pEF1a-PE5max-EGFP

LOCUS       pEF1a-PE5max-EGFP               13486 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 13486)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 22, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..13486
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(38..76)
                     /label=OL4014
     primer_bind     57..102
                     /label=OL4012
     promoter        87..1265
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     primer_bind     87..103
                     /label=OL2675
     primer_bind     87..103
                     /label=OL2712
     primer_bind     264..278
                     /label=OL3734
     intron          318..1256
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     340..357
                     /label=ef1a-seq1
     primer_bind     1116..1135
                     /label=ef1a-seq2
     primer_bind     1213..1233
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     complement(1236..1291)
                     /label=OL4011
     primer_bind     complement(1237..1325)
                     
     primer_bind     1270..1311
                     /label=OL4013
     primer_bind     1288..1320
                     /label=OL2387
     primer_bind     1304..1324
                     /label=OL3918
     primer_bind     1307..1326
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1307..1325
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1307..1325
                     /label=OL1124
     primer_bind     complement(1322..1341)
                     /label=OL1206
     primer_bind     complement(1326..1415)
                     /label=OL1393
     primer_bind     complement(1326..1338)
                     /label=OL3698
     primer_bind     1355..1369
                     /label=OL2802
     primer_bind     complement(1360..1374)
                     /label=OL2538
     primer_bind     complement(1373..1395)
                     /label=OL1036
     CDS             1375..1395
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     1376..1417
                     /label=OL1039
     primer_bind     1376..1415
                     /label=OL1296
     primer_bind     complement(1376..1395)
                     /label=OL1298
     CDS             1396..5496
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(R221K N394K H840A)
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRKLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLKREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     complement(1400..1419)
                     /label=OL1051
     primer_bind     1624..1648
                     /label=OL2715
     primer_bind     complement(1624..1648)
                     /label=OL2716
     primer_bind     complement(1844..1863)
                     /label=OL1336
     primer_bind     1864..1878
                     /label=OL1386
     primer_bind     complement(1868..1886)
                     /label=OL3674
     primer_bind     complement(1932..1951)
                     /label=OL989
     primer_bind     complement(2004..2014)
                     /label=OL900
     primer_bind     complement(2037..2066)
                     /label=OL1321
     primer_bind     2042..2071
                     /label=OL1322
     primer_bind     2204..2218
                     /label=OL1270
     primer_bind     complement(2557..2591)
                     /label=OL1323
     primer_bind     2557..2591
                     /label=OL1324
     primer_bind     2688..2707
                     /label=OL1447
     primer_bind     complement(2891..2910)
                     /label=OL1446
     primer_bind     3116..3130
                     /label=OL1271
     primer_bind     3506..3525
                     /label=OL2835
     primer_bind     3614..3648
                     /label=OL2384
     primer_bind     complement(3616..3648)
                     /label=OL1893
     primer_bind     3862..3876
                     /label=OL1272
     primer_bind     complement(3895..3926)
                     /label=OL1316
     primer_bind     3895..3926
                     /label=OL1317
     primer_bind     complement(3949..3968)
                     /label=OL1921
     primer_bind     4040..4056
                     /label=OL2390
     primer_bind     4969..4988
                     /label=OL2391
     primer_bind     5199..5218
                     /label=OL1328
     primer_bind     complement(5470..5511)
                     /label=OL2388
     CDS             5497..5598
                     /label=SGGSx2-sv40NLS-SGGSx2
                     /note="/created_by=chenp"
     primer_bind     5497..5519
                     /label=OL1326
     primer_bind     5536..5552
                     /label=OL1153
     primer_bind     complement(5544..5560)
                     /label=OL1154
     CDS             5554..5574
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     5575..5620
                     /label=OL2673
     CDS             5599..7629
                     /label=RT codon optimized Genscript
                     /note="/created_by=jessiedavis"
     primer_bind     complement(6211..6250)
                     /label=OL1894
     primer_bind     6293..6307
                     /label=OL1416
     primer_bind     6427..6445
                     /label=OL2947
     primer_bind     complement(6453..6471)
                     /label=OL1920
     primer_bind     complement(6482..6513)
                     /label=OL2389
     primer_bind     6632..6651
                     /label=OL2719
     primer_bind     complement(6632..6651)
                     /label=OL2720
     primer_bind     7328..7342
                     /label=OL1417
     primer_bind     7563..7585
                     /label=OL447
     primer_bind     7630..7651
                     /label=OL2671
     CDS             7675..7695
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7705..7731
                     /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     complement(7709..7731)
                     /label=OL2672
     primer_bind     complement(7727..7740)
                     /label=OL2803
     CDS             7741..7797
                     /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     7741..7797
                     /label=OL866
     primer_bind     complement(7741..7797)
                     /label=OL867
     primer_bind     7741..7767
                     /label=OL2251
     primer_bind     7741..7765
                     /label=OL899
     primer_bind     7741..7764
                     /label=PE001
     primer_bind     7741..7760
                     /label=OL3636
     primer_bind     7741..7757
                     /label=OL2806
     primer_bind     complement(7779..7798)
                     /label=PE002
     primer_bind     7788..7812
                     /label=OL3077
     CDS             7798..10053
                     /label=MLH1 del754-756 genscript codon
                     /note="/created_by=chenp"
                     /note="/modified_by=chenp"
     primer_bind     7852..7887
                     /label=OL532
     primer_bind     complement(8158..8196)
                     /label=OL531
     primer_bind     8251..8265
                     /label=OL1418
     primer_bind     9814..9833
                     /label=OL1414
     primer_bind     complement(9814..9833)
                     /label=OL1415
     primer_bind     10024..10078
                     /label=OL899
     primer_bind     10053..10077
                     /label=PE001
     CDS             10054..10110
                     /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     10054..10110
                     /label=OL866
     primer_bind     complement(10054..10110)
                     /label=OL867
     primer_bind     10054..10080
                     /label=OL2251
     primer_bind     10054..10073
                     /label=OL3636
     primer_bind     10054..10070
                     /label=OL2806
     primer_bind     10078..10131
                     /label=OL1687
     primer_bind     complement(10091..10129)
                     /label=OL308
     primer_bind     10092..10129
                     /label=OL309
     primer_bind     complement(10092..10118)
                     /label=PE002
     primer_bind     10102..10128
                     /label=PE003
     CDS             10111..10830
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     10111..10131
                     /label=OL1686
     primer_bind     10111..10131
                     /label=OL1704
     primer_bind     10111..10129
                     /label=OL3080
     primer_bind     complement(10332..10351)
                     /label=OL957
     primer_bind     complement(10493..10511)
                     /label=OL3733
     primer_bind     10542..10564
                     /label=OL3306
     primer_bind     10764..10784
                     /label=OL697
     primer_bind     10774..10788
                     /label=OL1937
     primer_bind     complement(10805..10830)
                     /label=OL174
     primer_bind     complement(10806..10860)
                     /label=OL900
     primer_bind     complement(10807..10831)
                     /label=PE004
     primer_bind     complement(10807..10830)
                     /label=OL3081
     primer_bind     complement(10807..10827)
                     /label=OL1833
     primer_bind     10808..10822
                     /label=OL2807
     primer_bind     complement(10811..10850)
                     /label=OL1205
     primer_bind     complement(10811..10850)
                     /label=OL1215
     primer_bind     complement(10812..10853)
                     /label=OL1146
     primer_bind     complement(10812..10827)
                     /label=OL2847
     primer_bind     complement(10813..10855)
                     /label=OL1038
     primer_bind     10831..10850
                     /label=OL1207
     primer_bind     10831..10845
                     /label=OL2537
     primer_bind     complement(10833..10850)
                     /label=OL2808
     primer_bind     10836..10859
                     /label=OL1035
     CDS             10836..10853
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     10836..10853
                     /label=OL1133
     primer_bind     complement(10836..10853)
                     /label=OL1152
     primer_bind     complement(10876..10893)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10882..11106
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     10882..10901
                     /label=OL1407
     primer_bind     complement(10887..10906)
                     /label=OL1410
     primer_bind     complement(10926..10943)
                     /label=OL698
     primer_bind     complement(10928..10947)
                     /label=OL1938
     primer_bind     complement(11087..11106)
                     /label=OL1684
     primer_bind     complement(11177..11193)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11177..11193)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11190..11212)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11201..11217
                     /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(11225..11255)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     11239..11258
                     /label=OL 3278
     protein_bind    11270..11291
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11408..11425)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     11518..11534
                     /label=OL3695
     primer_bind     11518..11534
                     /label=OL3920
     primer_bind     complement(11518..11534)
                     /label=OL3923
     primer_bind     complement(11518..11533)
                     /label=OL3924
     primer_bind     11535..11549
                     /label=OL3984
     rep_origin      complement(11579..12167)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11659..11678)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(11761..11780)
                     /label=OL1041
     primer_bind     complement(11894..11913)
                     /label=OL631
     primer_bind     11950..11969
                     /label=OL798
     primer_bind     12211..12230
                     /label=OL659
     CDS             complement(12338..13198)
                     /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     complement(12634..12649)
                     /label=OL3925
     primer_bind     12651..12665
                     /label=OL3926
     primer_bind     12795..12814
                     /label=OL1922
     primer_bind     12961..12980
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(12961..12980)
                     /label=OL1923
     primer_bind     13114..13133
                     /label=OL1033
     primer_bind     complement(13114..13133)
                     /label=OL1034
     promoter        complement(13199..13303)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     13277..13307
                     /label=OL 3277
     primer_bind     complement(13277..13306)
                     /label=OL 3274
     primer_bind     complement(13382..13401)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     13382..13401
                     /label=OL1919
ORIGIN
        1 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
       61 cgatgtacgg gccagatata cgcgttggct ccggtgcccg tcagtgggca gagcgcacat
      121 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa
      181 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
      241 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
      301 ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg
      361 gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc
      421 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt
      481 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt
      541 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat
      601 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc
      661 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca
      721 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc
      781 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg
      841 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc
      901 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac
      961 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt
     1021 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag
     1081 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag
     1141 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat
     1201 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt
     1261 cgtgaggttt agtgaaccgt cagatccgct agagatccgc ggccgctaat acgactcact
     1321 atagggagag ccgccaccat gaaacggaca gccgacggaa gcgagttcga gtcaccaaag
     1381 aagaagcgga aagtcgacaa gaagtacagc atcggcctgg acatcggcac caactctgtg
     1441 ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc
     1501 aacaccgacc ggcacagcat caagaagaac ctgatcggag ccctgctgtt cgacagcggc
     1561 gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag
     1621 aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc
     1681 ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac
     1741 cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac
     1801 cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg
     1861 gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc
     1921 gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc
     1981 gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg
     2041 agcaagagca gaaagctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc
     2101 ctgttcggaa acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc
     2161 gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac
     2221 aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg
     2281 tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc
     2341 ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa
     2401 gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag
     2461 aacggctacg ccggctacat tgacggcgga gccagccagg aagagttcta caagttcatc
     2521 aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaagagagag
     2581 gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg
     2641 ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac
     2701 cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc
     2761 aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg
     2821 aacttcgagg aagtggtgga caagggcgct tccgcccaga gcttcatcga gcggatgacc
     2881 aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag
     2941 tacttcaccg tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag
     3001 cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt caagaccaac
     3061 cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac
     3121 tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat
     3181 ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg
     3241 gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg
     3301 aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac
     3361 accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc
     3421 aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg
     3481 atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag
     3541 ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc
     3601 atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc
     3661 gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac
     3721 agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg
     3781 aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg
     3841 cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac
     3901 gatgtggacg ctatcgtgcc tcagagcttt ctgaaggacg actccatcga caacaaggtg
     3961 ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg
     4021 aagaagatga agaactactg gcggcagctg ctgaacgcca agctgattac ccagagaaag
     4081 ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc
     4141 atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac
     4201 tcccggatga acactaagta cgacgagaat gacaagctga tccgggaagt gaaagtgatc
     4261 accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc
     4321 gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc
     4381 ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac
     4441 gacgtgcgga agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac
     4501 ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag
     4561 atccggaagc ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt gtgggataag
     4621 ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa
     4681 aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc
     4741 gataagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcgacagc
     4801 cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa
     4861 ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag
     4921 aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc
     4981 aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct
     5041 gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg
     5101 tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag
     5161 ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc
     5221 tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag
     5281 caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc
     5341 aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagaggtac
     5401 accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac
     5461 gagacacgga tcgacctgtc tcagctggga ggtgactccg gcggaagctc tggtggcagc
     5521 aagcggaccg ccgacggctc tgaattcgag agccctaaga agaaaagaaa ggtgagcgga
     5581 ggctctagcg gcggaagcac cctgaacatt gaagacgagt atagactgca tgaaacaagc
     5641 aaggaacccg acgtgtccct gggctccacc tggctgtccg actttcccca ggcctgggcc
     5701 gagacaggag gaatgggcct ggccgtgcgg caggcacccc tgatcatccc tctgaaggcc
     5761 acctctacac ccgtgagcat caagcagtac cctatgtctc aggaggccag actgggcatc
     5821 aagcctcaca tccagaggct gctggaccag ggcatcctgg tgccatgcca gagcccctgg
     5881 aacacaccac tgctgcccgt gaagaagcca ggcaccaatg actatagacc cgtgcaggat
     5941 ctgagagagg tgaacaagag ggtggaggat atccacccca ccgtgcccaa cccttacaat
     6001 ctgctgtccg gcctgccccc ttctcaccag tggtatacag tgctggacct gaaggatgcc
     6061 ttcttttgtc tgagactgca ccctaccagc cagccactgt tcgcctttga gtggagggac
     6121 cctgagatgg gcatctctgg ccagctgacc tggacacgcc tgcctcaggg cttcaagaat
     6181 agcccaacac tgtttaacga ggccctgcac cgcgacctgg cagatttccg gatccagcac
     6241 ccagatctga tcctgctgca gtacgtggac gatctgctgc tggccgccac cagcgagctg
     6301 gattgccagc agggaacacg cgccctgctg cagaccctgg gaaacctggg atatagggca
     6361 tccgccaaga aggcccagat ctgtcagaag caggtgaagt acctgggcta tctgctgaag
     6421 gagggccaga gatggctgac agaggccagg aaggagacag tgatgggcca gccaacaccc
     6481 aagaccccaa gacagctgag ggagttcctg ggcaaagcag gattttgcag gctgttcatc
     6541 ccaggattcg cagagatggc agcacctctg tacccactga ccaagccggg caccctgttt
     6601 aattggggcc ctgaccagca gaaggcctat caggagatca agcaggccct gctgacagca
     6661 ccagccctgg gcctgccaga cctgaccaag cctttcgagc tgtttgtgga tgagaagcag
     6721 ggctacgcca agggcgtgct gacccagaag ctgggaccat ggagacggcc cgtggcctat
     6781 ctgtccaaga agctggaccc agtggcagca ggatggccac catgcctgag gatggtggca
     6841 gcaatcgccg tgctgacaaa ggatgccggc aagctgacca tgggacagcc actggtcatc
     6901 ctggcaccac acgcagtgga ggccctggtg aagcagcctc cagatcgctg gctgtctaac
     6961 gcccggatga cacactacca ggccctgctg ctggacaccg atcgcgtgca gtttggccct
     7021 gtggtggccc tgaatccagc caccctgctg cctctgccag aggagggcct gcagcacaac
     7081 tgtctggaca tcctggcaga ggcacacgga acaaggccag acctgaccga tcagcccctg
     7141 cctgacgccg atcacacatg gtataccgat ggaagctccc tgctgcagga gggccagagg
     7201 aaggcaggag cagcagtgac cacagagaca gaagtgatct gggccaaggc cctgccagca
     7261 ggcacatccg cccagcgggc cgagctgatc gccctgaccc aggccctgaa gatggccgag
     7321 ggcaagaagc tgaacgtgta cacagactcc agatatgcct tcgccaccgc acacatccac
     7381 ggagagatct acaggcgccg gggctggctg acctctgagg gcaaggagat caagaacaag
     7441 gatgagatcc tggccctgct gaaggccctg tttctgccca agcggctgag catcatccac
     7501 tgtcctggac accagaaggg acactccgcc gaggcaaggg gcaatcggat ggccgaccag
     7561 gccgccagaa aggctgctat tactgaaact cccgacactt ccactctgct gattgaaaac
     7621 tcctcccctt ctggcggctc aaaaagaacc gccgacggca gcgaattcga gtctcccaag
     7681 aagaagagga aagtcggctc tggccctgcc gctaagagag tgaagctgga cggaagcgga
     7741 gctactaact tcagcctgct gaagcaggct ggagacgtgg aggagaaccc tggacctagc
     7801 ttcgttgctg gagtcatccg gagactggac gagacagtgg tgaacagaat tgccgccggc
     7861 gaggtgatcc agagacctgc caatgcaata aaggagatga tcgagaactg tctggacgcc
     7921 aagtccacaa gcattcaggt gatcgtgaag gagggcggac tgaagctgat ccagatccaa
     7981 gacaacggca caggcatcag aaaggaagat ctggacatcg tgtgtgaacg gttcaccaca
     8041 tctaagctgc agtcttttga ggatctggcc tctatcagta cctacggctt cagaggcgag
     8101 gccctggcca gcatcagcca cgtggcccat gtgaccatca ccaccaaaac cgccgacggc
     8161 aaatgcgctt atcgcgctag ctacagcgac ggcaagctga aagccccgcc aaagccttgc
     8221 gccggcaacc agggtacaca gataacagtg gaggatctgt tctacaacat cgccacccgg
     8281 agaaaggccc tgaaaaatcc cagcgaggag tacggcaaga tcctggaagt cgtcggcaga
     8341 tactccgtgc acaacgccgg aatcagcttt agcgtaaaga agcagggaga aaccgtggcc
     8401 gatgtgcgca ccctgccaaa tgccagcacc gtggataaca tcagaagcat tttcggaaat
     8461 gccgtgtcca gagaactgat cgagatcggc tgcgaagata agaccctggc ttttaagatg
     8521 aacggctaca tcagcaacgc caattactct gtgaagaagt gcatctttct tctgttcatc
     8581 aaccacagac tggtggaaag caccagcctg cggaaagcca tcgagacagt gtacgccgcc
     8641 tacctgccta agaacaccca ccccttcctg tacctgagcc tcgagatcag ccctcagaac
     8701 gtggacgtca atgtgcatcc tacaaagcac gaggtgcact tcctgcacga ggaaagcatc
     8761 ctggaaagag tgcagcagca cattgagagc aagctgctgg gctctaacag cagcagaatg
     8821 tacttcacac agaccctgct gcctggcctg gccggcccct caggcgaaat ggttaagtcc
     8881 acaacctctc tgacctcatc cagcaccagc ggttcttccg ataaggtgta cgcccaccag
     8941 atggtgcgga ccgactctcg ggagcagaag ctggacgcct ttctgcaacc tctgagcaaa
     9001 cctctgagct ctcagcctca ggccatcgtg accgaggaca agacagatat ctcctccggc
     9061 cgtgccagac agcaggacga agaaatgctc gagctgccag ctcctgccga ggtggccgcc
     9121 aagaaccaga gcctggaggg agataccaca aagggcacca gcgaaatgag cgagaagcgg
     9181 ggccctacct ccagcaaccc cagaaaacgg caccgggagg acagcgacgt ggaaatggtg
     9241 gaggacgaca gcagaaagga aatgacagcc gcttgtaccc ctagaagaag aatcatcaac
     9301 ctgacctccg tgctgagcct gcaggaggag atcaacgagc agggccacga ggtgctgaga
     9361 gagatgctgc acaatcacag cttcgtgggc tgcgtgaacc ctcaatgggc cctggctcag
     9421 catcaaacaa agctgtacct gctgaacacc accaagctga gcgaagagct gttctaccag
     9481 atcctcatct acgacttcgc caacttcggc gtgctacgcc tgagcgagcc cgcccctctg
     9541 tttgacctgg ccatgctggc tctggatagc ccagaaagcg gctggacaga agaggacgga
     9601 cctaaagagg ggctggctga atacatcgtg gagttcctga agaaaaaggc cgagatgctg
     9661 gccgactact tttctctgga aatcgacgag gaaggcaacc tgatcggcct gcctctgctg
     9721 atcgataact acgtgcctcc cctggaaggc ctgcccatct tcatcctgag actggctaca
     9781 gaggtgaact gggacgagga aaaggaatgc ttcgagtctc tgagcaagga gtgcgccatg
     9841 ttctatagca tcagaaaaca gtacatctct gaagagagca ctctgtctgg ccagcagagt
     9901 gaagtgcccg gaagcatccc caacagctgg aagtggaccg tggaacacat cgtgtacaag
     9961 gccctgcgga gccacattct ccctcctaag cacttcaccg aggacggcaa catcctgcag
    10021 ctggccaacc tgcccgacct ttataaggtt ttcgctacta acttcagcct gctgaagcag
    10081 gctggagacg tggaggagaa ccctggacct atggtgagca agggcgagga gctgttcacc
    10141 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
    10201 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
    10261 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
    10321 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
    10381 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
    10441 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
    10501 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
    10561 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
    10621 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
    10681 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
    10741 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
    10801 actctcggca tggacgagct gtacaagtaa ccggtcatca tcaccatcac cattgagttt
    10861 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
    10921 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
    10981 agaaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
    11041 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
    11101 ctatggcttc tgaggcggaa agaaccagct ggggctcgat accgtcgacc tctagctaga
    11161 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
    11221 cacacaacat acgagccgga agcataaagt gtaaagccta gggtgcctaa tgagtgagct
    11281 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
    11341 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
    11401 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
    11461 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
    11521 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
    11581 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
    11641 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
    11701 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
    11761 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
    11821 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
    11881 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
    11941 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
    12001 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
    12061 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
    12121 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
    12181 tcttttctac ggggtctgac actcagtgga acgaaaactc acgttaaggg attttggtca
    12241 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
    12301 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
    12361 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
    12421 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
    12481 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
    12541 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
    12601 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
    12661 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
    12721 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
    12781 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
    12841 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
    12901 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
    12961 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
    13021 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
    13081 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
    13141 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
    13201 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
    13261 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
    13321 tgccacctga cgtcgacgga tcgggagatc gatctcccga tcccctaggg tctactctca
    13381 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
    13441 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggca
//