LOCUS       sequence-10346-e        6974 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6974)
  AUTHORS   Pastrana DV, Tolstov YL, Becker JC, Moore PS, Chang Y, Buck CB
  TITLE     Quantitation of human seroresponsiveness to Merkel cell 
            polyomavirus.
  JOURNAL   PLoS Pathog. 2009 Sep . 5(9):e1000578.
  PUBMED    19750217
REFERENCE   2  (bases 1 to 6974)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS 
            Pathog. 2009 Sep . 5(9):e1000578."
COMMENT     Sequence Label: pGwf
FEATURES             Location/Qualifiers
     source          1..6974
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(53..73)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        58..387
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      238..373
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      412..421
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(418..420,421..423,424..1137)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(418..420,421..423,424..1134)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(463..484)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(724..743)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1071..1092
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1162..1283
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1173..1192)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1227..1246
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1465..2643
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1696..2634
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2591..2611
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    2667..2791
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2816..2821,2822..2839,2840..2846)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2900..3559
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2966..2985)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3901..4206
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     4085..4104
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(4247..4371)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    4433..5021
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4486..4506)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4904..4915)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5063..5635
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
     rep_origin      5806..6394
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6295..6314
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        6480..6527
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        6519..6537
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ccctgcaggg cctgaaataa cctctgaaag aggaacttgg ttaggtacct gtggaatgtg
       61 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
      121 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
      181 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
      241 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
      301 atttatacag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
      361 ttttttggag gcctaggctt ttgcaaaaag cttgattggg atccaccggt cgccaccatg
      421 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
      481 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
      541 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
      601 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
      661 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
      721 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
      781 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
      841 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
      901 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
      961 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1021 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     1081 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc
     1141 ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     1201 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     1261 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     1321 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg
     1381 ataaggatcc gggctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     1441 tgcggtggag aagagcatgc gtgaggctcc ggtgcccgtc agtgggcaga gcgcacatcg
     1501 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg
     1561 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcccttt tcccgagggt
     1621 gggggagaac cgtatataag tgcagtagtc gctgtgaacg ttctttttcg caacgggttt
     1681 gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt
     1741 tatggccctt gcgtgccttg aattacttcc acctggctgc agtacgtgat tcttgatccc
     1801 gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt gcgcttaagg agccccttcg
     1861 cctcgtgctt gagttgaggc ctggcctggg cgctggggcc gccgcgtgcg aatctggtgg
     1921 caccttcgcg cctgtctcgc tgctttcgat aagtctctag ccatttaaaa tttttgatga
     1981 cctgctgcga cgcttttttt ctggcaagat agtcttgtaa atgcgggcca agatctgcac
     2041 actggtattt cggtttttgg ggccgcgggc ggcgacgggg cccgtgcgtc ccagcgcaca
     2101 tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa tcggacgggg gtagtctcaa
     2161 gctggccggc ctgctctggt gcctggcctc gcgccgccgt gtatcgcccc gccctgggcg
     2221 gcaaggctgg cccggtcggc accagttgcg tgagcggaaa gatggccgct tcccggccct
     2281 gctgcaggga gctcaaaatg gaggacgcgg cgctcgggag agcgggcggg tgagtcaccc
     2341 acacaaagga aaagggcctt tccgtcctca gccgtcgctt catgtgactc cacggagtac
     2401 cgggcgccgt ccaggcacct cgattagttc tcgagctttt ggagtacgtc gtctttaggt
     2461 tggggggagg ggttttatgc gatggagttt ccccacactg agtgggtgga gactgaagtt
     2521 aggccagctt ggcacttgat gtaattctcc ttggaatttg ccctttttga gtttggatct
     2581 tggttcattc tcaagcctca gacagtggtt caaagttttt ttcttccatt tcaggtgtcg
     2641 tgaggaattc tctagagctt gatcaaacaa gtttgtacaa aaaagctgaa cgagaaacgt
     2701 aaaatgatat aaatatcaat atattaaatt agattttgca taaaaaacag actacataat
     2761 actgtaaaac acaacatatc cagtcactat ggcggccgca ttaggcaccc caggctttac
     2821 actttatgct tccggctcgt ataatgtgtg gattttgagt taggatccgt cgagattttc
     2881 aggagctaag gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc
     2941 ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa
     3001 ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa
     3061 gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaattccg
     3121 tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt
     3181 tttccatgag caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg
     3241 gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt
     3301 ccctaaaggg tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac
     3361 cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg
     3421 caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc
     3481 cgtctgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga
     3541 gtggcagggc ggggcgtaat ctagaggatc cggcttacta aaagccagat aacagtatgc
     3601 gtatttgcgc gctgattttt gcggtataag aatatatact gatatgtata cccgaagtat
     3661 gtcaaaaaga ggtgtgctat gaagcagcgt attacagtga cagttgacag cgacagctat
     3721 cagttgctca aggcatatat gatgtcaata tctccggtct ggtaagcaca accatgcaga
     3781 atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa gggatggctg
     3841 aggtcgcccg gtttattgaa atgaacggct cttttgctga cgagaacagg gactggtgaa
     3901 atgcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta
     3961 cagagtgata ttattgacac gcccgggcga cggatggtga tccccctggc cagtgcacgt
     4021 ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc
     4081 tggcgcatga tgaccaccga tatggccagt gtgccggtct ccgttatcgg ggaagaagtg
     4141 gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga
     4201 atataaatgt caggctccgt tatacacagc cagtctgcag gtcgaccata gtgactggat
     4261 atgttgtgtt ttacagtatt atgtagtctg ttttttatgc aaaatctaat ttaatatatt
     4321 gatatttata tcattttacg tttctcgttc agctttcttg tacaaagtgg ttcgatctag
     4381 aatggctagt ggatcccccg ggctgcagga attcgatatc aagcttatcg ataatcaacc
     4441 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     4501 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     4561 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     4621 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     4681 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     4741 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     4801 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     4861 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     4921 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     4981 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gtcggcccac
     5041 tgctccctaa acctgagcta gcattatccc taatacctgc caccccactc ttaatcagtg
     5101 gtggaagaac ggtctcagaa ctgtttgttt caattggcca tttaagttta gtagtaaaag
     5161 actggttaat gataacaatg catcgtaaaa ccttcagaag gaaaggagaa tgttttgtgg
     5221 accactttgg ttttcttttt tgcgtgtggc agttttaagt tattagtttt taaaatcagt
     5281 actttttaat ggaaacaact tgaccaaaaa tttgtcacag aattttgaga cccattaaaa
     5341 aagttaaatg agaaacctgt gtgttccttt ggtcaacacc gagacattta ggtgaaagac
     5401 atctaattct ggttttacga atctggaaac ttcttgaaaa tgtaattctt gagttaacac
     5461 ttctgggtgg agaatagggt tgttttcccc ccacataatt ggaaggggaa ggaatatcat
     5521 ttaaagctat gggagggttt ctttgattac aacactggag agaaatgcag catgttgctg
     5581 attgcctgtc actaaaacag gccaaaaact gagtccttgg gttgcataga aagcttcatg
     5641 ttgctaaacc aatgttaagt gaatctttgg aaacaaaatg tttccaaatt actgggatgt
     5701 gcatgttgaa acgtgggtta attaactagc catgaccaaa atcccttaac gtgagttttc
     5761 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5821 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5881 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5941 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6001 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6061 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6121 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6181 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6241 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6301 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6361 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6421 gttcctggcc ttttgctggc cttttgctca catgttctta attaaatttt tcaaaagtag
     6481 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgactca ctataggagg
     6541 gccatcatgg ccaagttgac cagtgctgtc ccagtgctca cagccaggga tgtggctgga
     6601 gctgttgagt tctggactga caggttgggg ttctccagag attttgtgga ggatgacttt
     6661 gcaggtgtgg tcagagatga tgtcaccctg ttcatctcag cagtccagga ccaggtggtg
     6721 cctgacaaca ccctggcttg ggtgtgggtg agaggactgg atgagctgta tgctgagtgg
     6781 agtgaggtgg tctccaccaa cttcagggat gccagtggcc ctgccatgac agagattgga
     6841 gagcagccct gggggagaga gtttgccctg agagacccag caggcaactg tgtgcacttt
     6901 gtggcagagg agcaggactg aggataagaa ttgtaacaaa aaaccccgcc ccggcggggt
     6961 tttttgttaa ttaa
//
