pCMV-RT (PE6e)-EGFP

LOCUS      pCMV-RT (PE6e)-EGFP     5850 bp DNA     circular SYN 06-NOV-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5850)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 20, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..5850
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        87..466
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     101..128
                     /label=OL1151
     primer_bind     complement(106..130)
                     /label=OL1047
     primer_bind     complement(168..195)
                     /label=OL1815
     primer_bind     318..355
                     /label=OL1892
     primer_bind     388..407
                     /label=OL064
     promoter        467..670
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     569..588
                     /label=OL790
     primer_bind     615..636
                     /label=OL096
     primer_bind     621..641
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     621..640
                     /label=OL1335
     primer_bind     complement(622..639)
                     /label=OL400
     primer_bind     622..639
                     /label=OL443
     primer_bind     640..663
                     /label=OL152
     primer_bind     640..663
                     /label=OL159
     primer_bind     649..671
                     /label=OL084
     primer_bind     693..725
                     /label=OL2387
     primer_bind     712..731
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        712..730
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     712..730
                     /label=OL1124
     primer_bind     complement(727..746)
                     /label=OL1206
     primer_bind     complement(765..779)
                     /label=OL2538
     primer_bind     771..852
                     /label=OL1012
     primer_bind     complement(774..854)
                     /label=OL1013
     primer_bind     complement(778..824)
                     /label=OL2674
     primer_bind     complement(778..800)
                     /label=OL1036
     CDS             780..800
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     781..850
                     /label=OL1037
     primer_bind     complement(781..800)
                     /label=OL1298
     CDS             800
                     /label=SGGSx2-sv40NLS-SGGSx2
                     /note="/created_by=chenp"
     primer_bind     801..846
                     /label=OL2673
     CDS             825..2315
                     /label=RT codon optimized Genscript
                     /note="/created_by=jessiedavis"
     primer_bind     complement(1437..1476)
                     /label=OL1894
     primer_bind     1519..1533
                     /label=OL1416
     primer_bind     complement(1679..1697)
                     /label=OL1920
     primer_bind     complement(1708..1739)
                     /label=OL2389
     primer_bind     complement(2295..2340)
                     /label=OL2669
     primer_bind     2316..2337
                     /label=OL2671
     CDS             2361..2381
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2391..2417
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     primer_bind     2393..2442
                     /label=OL2670
     primer_bind     complement(2395..2417)
                     /label=OL2672
     primer_bind     2417..2442
                     /label=OL899
     primer_bind     2417..2441
                     /label=PE001
     CDS             2418..2474
                     /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     2418..2474
                     /label=OL866
     primer_bind     complement(2418..2474)
                     /label=OL867
     primer_bind     2418..2444
                     /label=OL2251
     primer_bind     2442..2495
                     /label=OL1687
     primer_bind     complement(2455..2493)
                     /label=OL308
     primer_bind     2456..2493
                     /label=OL309
     primer_bind     complement(2456..2482)
                     /label=PE002
     primer_bind     2466..2492
                     /label=PE003
     CDS             2475..3194
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     2475..2495
                     /label=OL1686
     primer_bind     2475..2495
                     /label=OL1704
     primer_bind     complement(2696..2715)
                     /label=OL957
     primer_bind     3128..3148
                     /label=OL697
     primer_bind     3138..3152
                     /label=OL1937
     primer_bind     complement(3169..3194)
                     /label=OL174
     primer_bind     complement(3170..3224)
                     /label=OL900
     primer_bind     complement(3171..3195)
                     /label=PE004
     primer_bind     complement(3171..3191)
                     /label=OL1833
     primer_bind     complement(3175..3214)
                     /label=OL1205
     primer_bind     complement(3175..3214)
                     /label=OL1215
     primer_bind     complement(3176..3217)
                     /label=OL1146
     primer_bind     complement(3177..3219)
                     /label=OL1038
     primer_bind     3191..3224
                     /label=OL1014
     primer_bind     3195..3214
                     /label=OL1207
     primer_bind     3195..3209
                     /label=OL2537
     primer_bind     3200..3223
                     /label=OL1035
     CDS             3200..3217
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     3200..3217
                     /label=OL1133
     primer_bind     complement(3200..3217)
                     /label=OL1152
     primer_bind     complement(3240..3257)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3246..3470
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal
                     牛生长激素聚腺苷酸化信号
                     牛生长激素（BGH）多聚腺苷酸化信号能够保证高效的转录终止和mRNA的poly A结构"
     primer_bind     3246..3265
                     /label=OL1407
     primer_bind     complement(3251..3270)
                     /label=OL1410
     primer_bind     complement(3290..3307)
                     /label=OL698
     primer_bind     complement(3292..3311)
                     /label=OL1938
     primer_bind     complement(3451..3470)
                     /label=OL1684
     primer_bind     complement(3541..3557)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3541..3557)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3554..3576)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3565..3581
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(3589..3619)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3634..3655
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3772..3789)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3943..4531)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4023..4042)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4125..4144)
                     /label=OL1041
     primer_bind     complement(4258..4277)
                     /label=OL631
     primer_bind     4314..4333
                     /label=OL798
     primer_bind     4575..4594
                     /label=OL659
     CDS             complement(4702..5562)
                     /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     5159..5178
                     /label=OL1922
     primer_bind     5325..5344
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(5325..5344)
                     /label=OL1923
     primer_bind     5478..5497
                     /label=OL1033
     primer_bind     complement(5478..5497)
                     /label=OL1034
     promoter        complement(5563..5667)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5746..5765)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5746..5765
                     /label=OL1919
ORIGIN
        1 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
       61 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
      121 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      181 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      241 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      301 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      361 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      421 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      481 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      541 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      601 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      661 aagcagagct ggtttagtga accgtcagat ccgctagaga tccgcggccg ctaatacgac
      721 tcactatagg gagagccgcc accatgaaac ggacagccga cggaagcgag ttcgagtcac
      781 caaagaagaa gcggaaagtc agcggaggct ctagcggcgg aagcaccctg aacattgaag
      841 acgagtatag actgcatgaa acaagcaagg aacccgacgt gtccctgggc tccacctggc
      901 tgtccgactt tccccaggcc tgggccgaga caggaggaat gggcctggcc gtgcggcagg
      961 cacccctgat catccctctg aaggccacct ctacacccgt gagcatcaag cagtacccta
     1021 tgtctcagga ggccagactg ggcatcaagc ctcacatcca gaggctgctg gaccagggca
     1081 tcctggtgcc atgccagagc ccctggaaca caccactgct gcccgtgaag aagccaggca
     1141 ccaatgacta tagacccgtg caggatctga gagaggtgaa caagagggtg gaggatatcc
     1201 accccaccgt gcccaaccct tacaatctgc tgtccggcct gcccccttct caccagtggt
     1261 atacagtgct ggacctgaag gatgccttct tttgtctgag actgcaccct accagccagc
     1321 cactgttcgc ctttgagtgg agggaccctg agatgggcat ctctggccag ctgacctgga
     1381 cacgcctgcc tcagggcttc aagaatagcc caacactgtt taacgaggcc ctgcaccgcg
     1441 acctggcaga tttccggatc cagcacccag atctgatcct gctgcagtac gtggacgatc
     1501 tgctgctggc cgccaccagc gagctggatt gccagcaggg aacacgcgcc ctgctgcaga
     1561 ccctgggaaa cctgggatat agggcatccg ccaagaaggc ccagatctgt cagaagcagg
     1621 tgaagtacct gggctatctg ctgaaggagg gccagagatg gctgacagag gccaggaagg
     1681 agacagtgat gggccagcca acacccaaga ccccaagaca gctgagggag ttcctgggca
     1741 aagcaggatt ttgcaggctg ttcatcccag gattcgcaga gatggcagca cctctgtacc
     1801 cactgaccaa gccgggcacc ctgtttaatt ggggccctga ccagcagaag gcctatcagg
     1861 agatcaagca ggccctgctg acagcaccag ccctgggcct gccagacctg accaagcctt
     1921 tcgagctgtt tgtggatgag aagcagggct acgccaaggg cgtgctgacc cagaagctgg
     1981 gaccatggag acggcccgtg gcctatctgt ccaagaagct ggacccagtg gcagcaggat
     2041 ggccaccatg cctgaggatg gtggcagcaa tcgccgtgct gacaaaggat gccggcaagc
     2101 tgaccatggg acagccactg gtcatcctgg caccacacgc agtggaggcc ctggtgaagc
     2161 agcctccaga tcgctggctg tctaacgccc ggatgacaca ctaccaggcc ctgctgctgg
     2221 acaccgatcg cgtgcagttt ggccctgtgg tggccctgaa tccagccacc ctgctgcctc
     2281 tgccagagga gggcctgcag cacaactgtc tggactctgg cggctcaaaa agaaccgccg
     2341 acggcagcga attcgagtct cccaagaaga agaggaaagt cggctctggc cctgccgcta
     2401 agagagtgaa gctggacgct actaacttca gcctgctgaa gcaggctgga gacgtggagg
     2461 agaaccctgg acctatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     2521 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     2581 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     2641 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     2701 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     2761 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     2821 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     2881 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     2941 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     3001 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     3061 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     3121 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     3181 agctgtacaa gtaaccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc
     3241 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     3301 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgagaaaa ttgcatcgca
     3361 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     3421 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     3481 ggaaagaacc agctggggct cgataccgtc gacctctagc tagagcttgg cgtaatcatg
     3541 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3601 cggaagcata aagtgtaaag cctagggtgc ctaatgagtg agctaactca cattaattgc
     3661 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3721 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3781 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3841 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3901 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3961 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4021 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4081 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4141 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4201 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4261 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4321 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4381 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4441 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4501 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4561 tgacactcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4621 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4681 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4741 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4801 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4861 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4921 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4981 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5041 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5101 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5161 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5221 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5281 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5341 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5401 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5461 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5521 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5581 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5641 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     5701 cggatcggga gatcgatctc ccgatcccct agggtctact ctcagtacaa tctgctctga
     5761 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     5821 cgcgagcaaa atttaagcta caacaaggca
//