pGL3-EM/T/G/ER/B/N/F/D/L/K-Nicking/PegRNA-PGK-puroR-mRFP1

LOCUS       pGL3-EM/T/G/ER/B/N/F/D/L/K-Nicking/PegRNA-PGK-puroR-mRFP1            9367 bp DNA     circular SYN 08-NOV-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9367)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 22, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..9367
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(21..44)
                     /label=OL459
     primer_bind     24..40
                     /label=OL2990
     primer_bind     24..40
                     /label=OL2992
     primer_bind     24..40
                     /label=OL2994
     primer_bind     24..40
                     /label=OL2996
     primer_bind     27..45
                     /label=OL2998
     primer_bind     37..61
                     /label=OL1451
     misc_feature    41..115
                     /label=hCtRNA
     primer_bind     56..79
                     /label=OL464
     primer_bind     complement(98..115)
                     /label=OL465
     misc_feature    116..135
                     /label=EMX1 nickingRNA
     misc_RNA        136..211
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     136..176
                     /label=OL462
     primer_bind     complement(194..230)
                     /label=OL463
     misc_feature    212..286
                     /label=hCtRNA
     primer_bind     212..232
                     /label=OL1451
     primer_bind     227..250
                     /label=OL464
     primer_bind     complement(269..286)
                     /label=OL465
     misc_feature    287..306
                     /label=EMX1 spacer
     misc_RNA        307..382
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     307..347
                     /label=OL462
     primer_bind     complement(365..382)
                     /label=OL463
     misc_feature    383..404
                     /label=RTT
     misc_feature    405..417
                     /label=PBS
     primer_bind     complement(421..450)
                     /label=OL2997
     primer_bind     complement(421..440)
                     /label=OL2995
     primer_bind     complement(424..443)
                     /label=OL2991
     primer_bind     complement(424..442)
                     /label=OL2989
     primer_bind     complement(424..441)
                     /label=OL2993
     primer_bind     436..457
                     /label=OL2990
     primer_bind     437..457
                     /label=OL2994
     primer_bind     437..457
                     /label=OL2996
     primer_bind     438..457
                     /label=OL2992
     primer_bind     444..462
                     /label=OL2998
     primer_bind     454..478
                     /label=OL1451
     misc_feature    458..532
                     /label=hCtRNA
     primer_bind     473..496
                     /label=OL464
     primer_bind     complement(515..535)
                     /label=OL465
     misc_feature    533..552
                     /label=TP53 nickingRNA
     primer_bind     550..593
                     /label=OL462
     misc_RNA        553..628
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(611..647)
                     /label=OL463
     misc_feature    629..703
                     /label=hCtRNA
     primer_bind     629..649
                     /label=OL1451
     primer_bind     644..667
                     /label=OL464
     primer_bind     complement(686..703)
                     /label=OL465
     misc_feature    704..723
                     /label=TP53 spacer
     misc_RNA        724..799
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     724..764
                     /label=OL462
     primer_bind     complement(782..799)
                     /label=OL463
     misc_feature    800..813
                     /label=RTT
     misc_feature    814..826
                     /label=PBS
     primer_bind     complement(830..859)
                     /label=OL2997
     primer_bind     complement(830..849)
                     /label=OL2995
     primer_bind     complement(833..852)
                     /label=OL2991
     primer_bind     complement(833..851)
                     /label=OL2989
     primer_bind     complement(833..850)
                     /label=OL2993
     primer_bind     845..866
                     /label=OL2990
     primer_bind     846..866
                     /label=OL2994
     primer_bind     846..866
                     /label=OL2996
     primer_bind     847..866
                     /label=OL2992
     primer_bind     853..871
                     /label=OL2998
     primer_bind     863..887
                     /label=OL1451
     misc_feature    867..941
                     /label=hCtRNA
     primer_bind     882..905
                     /label=OL464
     primer_bind     complement(924..941)
                     /label=OL465
     misc_feature    942..961
                     /label=GBA1 nickingRNA
     primer_bind     942..961
                     /label=OL3594
     misc_RNA        962..1037
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     962..1002
                     /label=OL462
     primer_bind     complement(1020..1056)
                     /label=OL463
     misc_feature    1038..1112
                     /label=hCtRNA
     primer_bind     1038..1058
                     /label=OL1451
     primer_bind     1053..1076
                     /label=OL464
     primer_bind     complement(1095..1112)
                     /label=OL465
     misc_feature    1113..1132
                     /label=GBA1 spacer
     misc_RNA        1133..1208
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     1133..1173
                     /label=OL462
     primer_bind     complement(1191..1208)
                     /label=OL463
     misc_feature    1209..1225
                     /label=RTT
     misc_feature    1226..1238
                     /label=PBS
     primer_bind     complement(1242..1271)
                     /label=OL2997
     primer_bind     complement(1242..1261)
                     /label=OL2995
     primer_bind     complement(1245..1264)
                     /label=OL2991
     primer_bind     complement(1245..1263)
                     /label=OL2989
     primer_bind     complement(1245..1262)
                     /label=OL2993
     primer_bind     1257..1278
                     /label=OL2990
     primer_bind     1258..1278
                     /label=OL2994
     primer_bind     1258..1278
                     /label=OL2996
     primer_bind     1259..1278
                     /label=OL2992
     primer_bind     1265..1283
                     /label=OL2998
     primer_bind     1275..1299
                     /label=OL1451
     misc_feature    1279..1353
                     /label=hCtRNA
     primer_bind     1294..1317
                     /label=OL464
     primer_bind     complement(1336..1353)
                     /label=OL465
     misc_feature    1354..1373
                     /label=ERCC6 nickingRNA
     misc_RNA        1374..1449
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     1374..1414
                     /label=OL462
     primer_bind     complement(1432..1468)
                     /label=OL463
     misc_feature    1450..1524
                     /label=hCtRNA
     primer_bind     1450..1470
                     /label=OL1451
     primer_bind     1465..1488
                     /label=OL464
     primer_bind     complement(1507..1525)
                     /label=OL465
     misc_feature    1525..1544
                     /label=ERCC6 spacer
     misc_RNA        1545..1620
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     1545..1585
                     /label=OL462
     primer_bind     complement(1603..1621)
                     /label=OL463
     misc_feature    1621..1635
                     /label=RTT
     misc_feature    1635
                     /label=a to g
     misc_feature    1636..1653
                     /label=PBS
     primer_bind     complement(1657..1686)
                     /label=OL2997
     primer_bind     complement(1657..1676)
                     /label=OL2995
     primer_bind     complement(1660..1679)
                     /label=OL2991
     primer_bind     complement(1660..1678)
                     /label=OL2989
     primer_bind     complement(1660..1677)
                     /label=OL2993
     primer_bind     1672..1693
                     /label=OL2990
     primer_bind     1673..1693
                     /label=OL2994
     primer_bind     1673..1693
                     /label=OL2996
     primer_bind     1674..1693
                     /label=OL2992
     primer_bind     1680..1698
                     /label=OL2998
     primer_bind     1690..1714
                     /label=OL1451
     misc_feature    1694..1768
                     /label=hCtRNA
     primer_bind     1709..1732
                     /label=OL464
     primer_bind     complement(1751..1769)
                     /label=OL465
     primer_bind     1766..1788
                     /label=OL193
     misc_feature    1769..1788
                     /label=BAG3 nickingRNA
     primer_bind     1769..1788
                     /label=OL3011
     primer_bind     1788..1829
                     /label=OL462
     misc_RNA        1789..1864
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(1847..1883)
                     /label=OL463
     misc_feature    1865..1939
                     /label=hCtRNA
     primer_bind     1865..1885
                     /label=OL1451
     primer_bind     1880..1903
                     /label=OL464
     primer_bind     complement(1922..1939)
                     /label=OL465
     misc_feature    1940..1959
                     /label=BAG3 spacer
     misc_RNA        1960..2035
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     1960..2000
                     /label=OL462
     primer_bind     complement(2018..2036)
                     /label=OL463
     misc_feature    2036..2047
                     /label=RTT
     misc_feature    2048..2062
                     /label=PBS
     primer_bind     complement(2066..2095)
                     /label=OL2997
     primer_bind     complement(2066..2085)
                     /label=OL2995
     primer_bind     complement(2069..2088)
                     /label=OL2991
     primer_bind     complement(2069..2087)
                     /label=OL2989
     primer_bind     complement(2069..2086)
                     /label=OL2993
     primer_bind     2081..2102
                     /label=OL2990
     primer_bind     2082..2102
                     /label=OL2994
     primer_bind     2082..2102
                     /label=OL2996
     primer_bind     2083..2102
                     /label=OL2992
     primer_bind     2089..2107
                     /label=OL2998
     primer_bind     2099..2123
                     /label=OL1451
     misc_feature    2103..2177
                     /label=hCtRNA
     primer_bind     2118..2141
                     /label=OL464
     primer_bind     complement(2160..2178)
                     /label=OL465
     misc_feature    2178..2197
                     /label=NMB nickingRNA
     primer_bind     2178..2197
                     /label=OL3012
     primer_bind     2197..2238
                     /label=OL462
     misc_RNA        2198..2273
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(2256..2292)
                     /label=OL463
     misc_feature    2274..2348
                     /label=hCtRNA
     primer_bind     2274..2294
                     /label=OL1451
     primer_bind     2289..2312
                     /label=OL464
     primer_bind     complement(2331..2349)
                     /label=OL465
     misc_feature    2349..2368
                     /label=NMB spacer
     misc_RNA        2369..2444
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     2369..2409
                     /label=OL462
     primer_bind     complement(2427..2447)
                     /label=OL463
     misc_feature    2445..2458
                     /label=RTT
     misc_feature    2459..2469
                     /label=PBS
     primer_bind     complement(2473..2502)
                     /label=OL2997
     primer_bind     complement(2473..2492)
                     /label=OL2995
     primer_bind     complement(2476..2495)
                     /label=OL2991
     primer_bind     complement(2476..2494)
                     /label=OL2989
     primer_bind     complement(2476..2493)
                     /label=OL2993
     primer_bind     2488..2509
                     /label=OL2990
     primer_bind     2489..2509
                     /label=OL2994
     primer_bind     2489..2509
                     /label=OL2996
     primer_bind     2490..2509
                     /label=OL2992
     primer_bind     2496..2514
                     /label=OL2998
     primer_bind     2506..2530
                     /label=OL1451
     misc_feature    2510..2584
                     /label=hCtRNA
     primer_bind     2525..2548
                     /label=OL464
     primer_bind     complement(2567..2584)
                     /label=OL465
     primer_bind     2582..2604
                     /label=OL2094
     misc_feature    2585..2604
                     /label=FANCF-nickingRNA
     primer_bind     2604..2645
                     /label=OL462
     misc_RNA        2605..2680
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(2663..2699)
                     /label=OL463
     misc_feature    2681..2755
                     /label=hCtRNA
     primer_bind     2681..2701
                     /label=OL1451
     primer_bind     2696..2719
                     /label=OL464
     primer_bind     complement(2738..2755)
                     /label=OL465
     misc_feature    2756..2775
                     /label=FANCF-spacer
     misc_RNA        2776..2851
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     2776..2816
                     /label=OL462
     primer_bind     complement(2834..2854)
                     /label=OL463
     misc_feature    2852..2868
                     /label=RT template
     misc_feature    2864
                     /label=G to T
     misc_feature    2869..2881
                     /label=PBS
     primer_bind     complement(2885..2914)
                     /label=OL2997
     primer_bind     complement(2885..2904)
                     /label=OL2995
     primer_bind     complement(2888..2907)
                     /label=OL2991
     primer_bind     complement(2888..2906)
                     /label=OL2989
     primer_bind     complement(2888..2905)
                     /label=OL2993
     primer_bind     2900..2921
                     /label=OL2990
     primer_bind     2901..2921
                     /label=OL2994
     primer_bind     2901..2921
                     /label=OL2996
     primer_bind     2902..2921
                     /label=OL2992
     primer_bind     2908..2926
                     /label=OL2998
     primer_bind     2918..2942
                     /label=OL1451
     misc_feature    2922..2996
                     /label=hCtRNA
     primer_bind     2937..2960
                     /label=OL464
     primer_bind     complement(2979..2996)
                     /label=OL465
     misc_feature    2997..3017
                     /label=DNMT1-nickingRNA
     misc_RNA        3018..3093
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     3018..3058
                     /label=OL462
     primer_bind     complement(3076..3112)
                     /label=OL463
     misc_feature    3094..3168
                     /label=hCtRNA
     primer_bind     3094..3114
                     /label=OL1451
     primer_bind     3109..3132
                     /label=OL464
     primer_bind     complement(3151..3168)
                     /label=OL465
     misc_feature    3169..3188
                     /label=DNMT1 spacer
     misc_RNA        3189..3264
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     3189..3229
                     /label=OL462
     primer_bind     complement(3247..3264)
                     /label=OL463
     misc_feature    3265..3275
                     /label=RT template
     misc_feature    3271
                     /label=C352A
     misc_feature    3276..3288
                     /label=PBS
     primer_bind     complement(3280..3297)
                     /label=OL3000
     primer_bind     complement(3292..3321)
                     /label=OL2997
     primer_bind     complement(3292..3311)
                     /label=OL2995
     primer_bind     complement(3295..3314)
                     /label=OL2991
     primer_bind     complement(3295..3313)
                     /label=OL2989
     primer_bind     complement(3295..3312)
                     /label=OL2993
     primer_bind     3307..3328
                     /label=OL2990
     primer_bind     3308..3328
                     /label=OL2994
     primer_bind     3308..3328
                     /label=OL2996
     primer_bind     3309..3328
                     /label=OL2992
     primer_bind     3315..3333
                     /label=OL2998
     primer_bind     3325..3349
                     /label=OL1451
     misc_feature    3329..3403
                     /label=hCtRNA
     primer_bind     3344..3367
                     /label=OL464
     primer_bind     complement(3386..3403)
                     /label=OL465
     misc_feature    3404..3423
                     /label=LRRK2-nickingRNA
     primer_bind     3404..3423
                     /label=OL3595
     misc_RNA        3424..3499
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     3424..3464
                     /label=OL462
     primer_bind     complement(3482..3518)
                     /label=OL463
     misc_feature    3500..3574
                     /label=hCtRNA
     primer_bind     3500..3520
                     /label=OL1451
     primer_bind     3515..3538
                     /label=OL464
     primer_bind     complement(3557..3574)
                     /label=OL465
     misc_feature    3575..3594
                     /label=LRKK2-Spacer
     misc_RNA        3595..3670
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     3595..3635
                     /label=OL462
     primer_bind     complement(3653..3672)
                     /label=OL463
     misc_feature    3671..3683
                     /label=RTT
     misc_feature    3684..3697
                     /label=PBS
     primer_bind     complement(3701..3730)
                     /label=OL2997
     primer_bind     complement(3701..3720)
                     /label=OL2995
     primer_bind     complement(3704..3723)
                     /label=OL2991
     primer_bind     complement(3704..3722)
                     /label=OL2989
     primer_bind     complement(3704..3721)
                     /label=OL2993
     primer_bind     3716..3737
                     /label=OL2990
     primer_bind     3717..3737
                     /label=OL2994
     primer_bind     3717..3737
                     /label=OL2996
     primer_bind     3718..3737
                     /label=OL2992
     primer_bind     3724..3742
                     /label=OL2998
     primer_bind     3734..3758
                     /label=OL1451
     misc_feature    3738..3812
                     /label=hCtRNA
     primer_bind     3753..3776
                     /label=OL464
     primer_bind     complement(3795..3812)
                     /label=OL465
     misc_feature    3813..3832
                     /label=KRAS-nickingRNA
     primer_bind     3832..3873
                     /label=OL462
     misc_RNA        3833..3908
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(3891..3927)
                     /label=OL463
     misc_feature    3909..3983
                     /label=hCtRNA
     primer_bind     3909..3929
                     /label=OL1451
     primer_bind     3924..3947
                     /label=OL464
     primer_bind     complement(3966..3983)
                     /label=OL465
     misc_feature    3984..4003
                     /label=KRAS spacer
     misc_RNA        4004..4079
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     4004..4044
                     /label=OL462
     primer_bind     complement(4062..4080)
                     /label=OL463
     misc_feature    4080..4091
                     /label=RTT
     primer_bind     complement(4092..4109)
                     /label=OL3582
     misc_feature    4092..4100
                     /label=PBS
     primer_bind     complement(4104..4123)
                     /label=OL2995
     primer_bind     complement(4104..4123)
                     /label=OL2997
     primer_bind     4106..4125
                     /label=OL458
     primer_bind     complement(4107..4123)
                     /label=OL2989
     primer_bind     complement(4107..4123)
                     /label=OL2991
     primer_bind     complement(4107..4123)
                     /label=OL2993
     primer_bind     complement(4113..4132)
                     /label=OL1916
     primer_bind     complement(4127..4143)
                     /label=OL1168
     misc_feature    4144..4261
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     4309..4329
                     /label=OL2290
     promoter        4310..4820
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4404..4423)
                     /label=OL1068
     primer_bind     complement(4404..4423)
                     /label=AG019_OL1068
     primer_bind     4594..4610
                     /label=OL446
     primer_bind     4739..4757
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             4842..5438
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4842..4861)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5338..5358
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     5409..5463
                     /label=OL901
     primer_bind     complement(5415..5438)
                     /label=OL306
     primer_bind     complement(5424..5446)
                     /label=OL384
     primer_bind     5431..5462
                     /label=OL379
     CDS             5439..5495
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     5439..5495
                     /label=OL866
     primer_bind     complement(5439..5495)
                     /label=OL867
     primer_bind     5476..5519
                     /label=OL902
     primer_bind     complement(5476..5503)
                     /label=OL966
     RBS             5478..5485
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             5496..6173
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(6151..6198)
                     /label=OL903
     primer_bind     complement(6155..6173)
                     /label=OL2976
     primer_bind     6172..6190
                     /label=OL965
     primer_bind     6174..6196
                     /label=OL305
     polyA_signal    6320..6441
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6331..6350)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6385..6404
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6630..6653)
                     /label=OL1043
     primer_bind     6631..6653
                     /label=OL1044
     primer_bind     complement(6689..6706)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6860..7448)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6940..6959)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(7042..7061)
                     /label=OL1041
     primer_bind     complement(7175..7194)
                     /label=OL631
     primer_bind     7231..7250
                     /label=OL798
     primer_bind     complement(7472..7511)
                     /label=OL660
     primer_bind     7492..7511
                     /label=OL659
     CDS             complement(7619..8479)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8242..8261
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     8395..8414
                     /label=OL1033
     primer_bind     complement(8395..8414)
                     /label=OL1034
     promoter        complement(8480..8584)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8611..9066
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8636..8645)
                     /label=OL305
     primer_bind     complement(8698..8717)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8908..8929
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    9197..9245
                     /note="synthetic polyadenylation signal"
     misc_feature    9259..9350
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     9259..9275
                     /label=OL1167
     primer_bind     9299..9318
                     /label=AG020_RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 agtgaacgga tctcgacggt atcgatcacg agactagcct agagggggta tagctcagtg
       61 gtagagcatt tgactgcaga tcaagaggtc cccggttcaa atccgggtgc cccctctccc
      121 cattggcctg cttcggtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
      181 tcaacttgaa aaagtggcac cgagtcggtg cagagggggt atagctcagt ggtagagcat
      241 ttgactgcag atcaagaggt ccccggttca aatccgggtg ccccctgagt ccgagcagaa
      301 gaagaagttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
      361 aaaagtggca ccgagtcggt gcgttgatgt gatgggagcc gttcttcttc tgctcggttt
      421 ttttctcgac ctcgagacaa gatcacgaga ctagcctaga gggggtatag ctcagtggta
      481 gagcatttga ctgcagatca agaggtcccc ggttcaaatc cgggtgcccc ctatcagagg
      541 cctggggacc ctgttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca
      601 acttgaaaaa gtggcaccga gtcggtgcag agggggtata gctcagtggt agagcatttg
      661 actgcagatc aagaggtccc cggttcaaat ccgggtgccc cctagcacat gacggaggtt
      721 gtggttttag agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa
      781 agtggcaccg agtcggtgcg ggcagtgcct cacaacctcc gtcatgtttt tttctcgacc
      841 tcgagacaag atcacgagac tagcctagag ggggtatagc tcagtggtag agcatttgac
      901 tgcagatcaa gaggtccccg gttcaaatcc gggtgccccc tacccttacc tacactctct
      961 ggttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag
     1021 tggcaccgag tcggtgcaga gggggtatag ctcagtggta gagcatttga ctgcagatca
     1081 agaggtcccc ggttcaaatc cgggtgcccc ctagccgacc acatggtaca gggttttaga
     1141 gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga
     1201 gtcggtgctt accctagagc ctcctgtacc atgtggtctt tttttctcga cctcgagaca
     1261 agatcacgag actagcctag agggggtata gctcagtggt agagcatttg actgcagatc
     1321 aagaggtccc cggttcaaat ccgggtgccc cctgtcagtg attagtggaa atggttttag
     1381 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
     1441 agtcggtgca gagggggtat agctcagtgg tagagcattt gactgcagat caagaggtcc
     1501 ccggttcaaa tccgggtgcc cccttcaatg ctttcatcac cagagtttta gagctagaaa
     1561 tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc
     1621 acttctatgc catccggtga tgaaagcatt gatttttttt ctcgacctcg agacaagatc
     1681 acgagactag cctagagggg gtatagctca gtggtagagc atttgactgc agatcaagag
     1741 gtccccggtt caaatccggg tgcccccttc tccttacctc aggtccgtgt tttagagcta
     1801 gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
     1861 gtgcagaggg ggtatagctc agtggtagag catttgactg cagatcaaga ggtccccggt
     1921 tcaaatccgg gtgcccccta ctcagccaga taaacagtgg ttttagagct agaaatagca
     1981 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcacctg
     2041 tccacgctgt ttatctggct gatttttttc tcgacctcga gacaagatca cgagactagc
     2101 ctagaggggg tatagctcag tggtagagca tttgactgca gatcaagagg tccccggttc
     2161 aaatccgggt gcccccttcc tttccaggtc acttcatgtt ttagagctag aaatagcaag
     2221 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcagagggg
     2281 gtatagctca gtggtagagc atttgactgc agatcaagag gtccccggtt caaatccggg
     2341 tgcccccttg gtccctcagg gaggtgtggt tttagagcta gaaatagcaa gttaaaataa
     2401 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgcggacag ctacccacac
     2461 ctccctgagt ttttttctcg acctcgagac aagatcacga gactagccta gagggggtat
     2521 agctcagtgg tagagcattt gactgcagat caagaggtcc ccggttcaaa tccgggtgcc
     2581 ccctggggtc ccaggtgctg acgtgtttta gagctagaaa tagcaagtta aaataaggct
     2641 agtccgttat caacttgaaa aagtggcacc gagtcggtgc agagggggta tagctcagtg
     2701 gtagagcatt tgactgcaga tcaagaggtc cccggttcaa atccgggtgc cccctggaat
     2761 cccttctgca gcaccgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
     2821 tcaacttgaa aaagtggcac cgagtcggtg cggaaaagcg atcaaggtgc tgcagaaggg
     2881 atttttttct cgacctcgag acaagatcac gagactagcc tagagggggt atagctcagt
     2941 ggtagagcat ttgactgcag atcaagaggt ccccggttca aatccgggtg ccccctgccc
     3001 ttcagctaaa ataaagggtt ttagagctag aaatagcaag ttaaaataag gctagtccgt
     3061 tatcaacttg aaaaagtggc accgagtcgg tgcagagggg gtatagctca gtggtagagc
     3121 atttgactgc agatcaagag gtccccggtt caaatccggg tgccccctga ttcctggtgc
     3181 cagaaacagt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
     3241 gaaaaagtgg caccgagtcg gtgcgtcacc actgtttctg gcaccaggtt tttttctcga
     3301 cctcgagaca agatcacgag actagcctag agggggtata gctcagtggt agagcatttg
     3361 actgcagatc aagaggtccc cggttcaaat ccgggtgccc cctgacagac ctgatcacct
     3421 accgttttag agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa
     3481 agtggcaccg agtcggtgca gagggggtat agctcagtgg tagagcattt gactgcagat
     3541 caagaggtcc ccggttcaaa tccgggtgcc ccctattgca aagattgctg actagtttta
     3601 gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc
     3661 gagtcggtgc agcaatgctg tagtcagcaa tctttgcttt ttttctcgac ctcgagacaa
     3721 gatcacgaga ctagcctaga gggggtatag ctcagtggta gagcatttga ctgcagatca
     3781 agaggtcccc ggttcaaatc cgggtgcccc ctacaagatt tacctctatt gtgttttaga
     3841 gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga
     3901 gtcggtgcag agggggtata gctcagtggt agagcatttg actgcagatc aagaggtccc
     3961 cggttcaaat ccgggtgccc cctaaacttg tggtagttgg agcgttttag agctagaaat
     4021 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgca
     4081 cgccatcagc tccaactacc tttttttctc gacctcgaga caaatggcag tattcatcca
     4141 caattttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     4201 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     4261 tcgggtttat tacagggaca gcagagatcc actttggccg cggctcgagg gggttggggt
     4321 tgcgcctttt ccaaggcagc cctgggtttg cgcagggacg cggctgctct gggcgtggtt
     4381 ccgggaaacg cagcggcgcc gaccctggga ctcgcacatt cttcacgtcc gttcgcagcg
     4441 tcacccggat cttcgccgct acccttgtgg gccccccggc gacgcttcct gctccgcccc
     4501 taagtcggga aggttccttg cggttcgcgg cgtgccggac gtgacaaacg gaagccgcac
     4561 gtctcactag taccctcgca gacggacagc gccagggagc aatggcagcg cgccgaccgc
     4621 gatgggctgt ggccaatagc ggctgctcag cagggcgcgc cgagagcagc ggccgggaag
     4681 gggcggtgcg ggaggcgggg tgtggggcgg tagtgtgggc cctgttcctg cccgcgcggt
     4741 gttccgcatt ctgcaagcct ccggagcgca cgtcggcagt cggctccctc gttgaccgaa
     4801 tcaccgacct ctctccccag ggggatccac cggagcttac catgaccgag tacaagccca
     4861 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt
     4921 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca
     4981 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     5041 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     5101 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     5161 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     5221 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     5281 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggaaacctcc gcgccccgca
     5341 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     5401 gaccgcgcac ctggtgcatg acccgcaagc ccggtgccgc tactaacttc agcctgctga
     5461 agcaggctgg agacgtggag gagaaccctg gacctatggc gagtagcgaa gacgttatca
     5521 aagagttcat gcgtttcaaa gttcgtatgg aaggttccgt taacggtcac gagttcgaaa
     5581 tcgaaggtga aggtgaaggt cgtccgtacg aaggtaccca gaccgctaaa ctgaaagtta
     5641 ccaaaggtgg tccgctgccg ttcgcttggg acatcctgtc cccgcagttc cagtacggtt
     5701 ccaaagctta cgttaaacac ccggctgaca tcccggacta cctgaaactg tccttcccgg
     5761 aaggtttcaa atgggaacgt gttatgaact tcgaagacgg tggtgttgtt accgttaccc
     5821 aggactcctc cctgcaagac ggtgagttca tctacaaagt taaactgcgt ggtaccaact
     5881 tcccgtccga cggtccggtt atgcagaaaa aaaccatggg ttgggaagct tccaccgaac
     5941 gtatgtaccc ggaagacggt gctctgaaag gtgaaatcaa aatgcgtctg aaactgaaag
     6001 acggtggtca ctacgacgct gaagttaaaa ccacctacat ggctaaaaaa ccggttcagc
     6061 tgccgggtgc ttacaaaacc gacatcaaac tggacatcac ctcccacaac gaagactaca
     6121 ccatcgttga acagtacgaa cgtgctgaag gtcgtcactc caccggtgct taacgcccgc
     6181 cccacgaccc gcagcgcccg accgaaagga gcgcacgacc ccatgcatcg gtacctttaa
     6241 gaccaatgac ttacaaggca gctgtagatc ttagccactt tctagagtcg gggcggccgg
     6301 ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     6361 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     6421 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     6481 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat
     6541 aaggatccgt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc
     6601 gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg
     6661 acaggtgccg gcagcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6721 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6781 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6841 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6901 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6961 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     7021 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     7081 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     7141 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     7201 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     7261 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     7321 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7381 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     7441 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     7501 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7561 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7621 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7681 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7741 gtgctgcaat gataccgcgg gacccacgct caccggctcc agatttatca gcaataaacc
     7801 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7861 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7921 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7981 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     8041 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     8101 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     8161 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     8221 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     8281 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     8341 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     8401 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     8461 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     8521 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     8581 cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg
     8641 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     8701 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     8761 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     8821 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     8881 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     8941 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     9001 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     9061 caatttgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     9121 cttcgctatt acgccagccc aagctaccat gataagtaag taatattaag gtacgggagg
     9181 tacttggagc ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt
     9241 gtgtgaatcg atagtactaa catacgctct ccatcaaaac aaaacgaaac aaaacaaact
     9301 agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga acatttctct atcgataggt
     9361 accgatt
//