LOCUS       sequence-10344-e        6610 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 6610)
  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 6610)
  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: pwM
FEATURES             Location/Qualifiers
     source          1..6610
                     /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..2691
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      2698..2707
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3983..4007)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    4069..4657
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4122..4142)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4540..4551)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4699..5271
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
     rep_origin      5442..6030
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5931..5950
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        6116..6163
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        6155..6173
                     /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 aaaagcaggc ttccggagcc
     2701 accatggccc cgaagcgcaa ggccagcagt acatgcaaga ccccaaagcg ccagtgcatc
     2761 ccgaaacccg gctgttgtcc caacgtcgca agcgtcccga agctgttggt caagggcggc
     2821 gtcgaggtgc tgagcgtcgt caccggcgag gactcaatca cacaaatcga actctacctc
     2881 aacccacgca tgggcgtcaa cagtcccgac ttgcccacca ccagcaattg gtacacctac
     2941 acctacgatc tgcaacccaa aggcagcagc ccggaccagc caatcaaaga gaacctgccc
     3001 gcctattccg tcgcccgcgt cagcctgccc atgttgaacg aagacatcac atgcgatacc
     3061 ctgcaaatgt gggaagccat cagcgtcaag accgaggtgg tcggcatcag cagcctgatc
     3121 aacgtccact actgggatat gaagcgcgtc cacgactacg gcgccggcat cccggtcagc
     3181 ggcgtcaact atcacatgtt cgcaatcggc ggcgagccgt tggacctgca agggctggtc
     3241 ctggactatc agacccagta ccccaagacc accaacggcg gccccatcac catcgagacc
     3301 gtgctcggcc gcaagatgac cccgaagaac caggggttgg acccacaagc caaggccaag
     3361 ctggacaagg acggcaacta ccccatcgag gtctggtgcc ccgacccgag caagaacgag
     3421 aattcccgct actacggcag catccaaacc gggagccaga ccccgaccgt gttgcagttc
     3481 agcaacaccc tgaccaccgt gctgctggac gaaaacggcg tcggcccact gtgcaagggc
     3541 gatggcctgt tcatctcatg cgccgatatc gtcggattcc tgttcaagac atccggcaag
     3601 atggccttgc acggcctgcc acgctacttc aacgtcaccc tgcgaaagcg ctgggtcaag
     3661 aatccatatc ccgtggtcaa cctgatcaat agcttgttca gcaatctgat gcccaaggtc
     3721 agcggccagc ccatggaggg caaggacaac caagtggagg aagtccgcat ctacgaaggc
     3781 agcgagcagc tgcccggcga ccccgacatc gtgcggttcc tggacaagtt cggccaagaa
     3841 aagaccgtct atcccaaacc aagcgtcgca cccgccgccg tgacattcca gtccaaccag
     3901 caagacaaag gcaaggcccc gctgaagggc ccgcaaaagg caagccagaa ggagtcacag
     3961 acgcaggagc tatgagccta ggacccagct ttcttgtaca aagtggttcg atctagaatg
     4021 gctagtggat cccccgggct gcaggaattc gatatcaagc ttatcgataa tcaacctctg
     4081 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     4141 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     4201 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     4261 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     4321 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     4381 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     4441 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
     4501 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     4561 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     4621 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg gcccactgct
     4681 ccctaaacct gagctagcat tatccctaat acctgccacc ccactcttaa tcagtggtgg
     4741 aagaacggtc tcagaactgt ttgtttcaat tggccattta agtttagtag taaaagactg
     4801 gttaatgata acaatgcatc gtaaaacctt cagaaggaaa ggagaatgtt ttgtggacca
     4861 ctttggtttt cttttttgcg tgtggcagtt ttaagttatt agtttttaaa atcagtactt
     4921 tttaatggaa acaacttgac caaaaatttg tcacagaatt ttgagaccca ttaaaaaagt
     4981 taaatgagaa acctgtgtgt tcctttggtc aacaccgaga catttaggtg aaagacatct
     5041 aattctggtt ttacgaatct ggaaacttct tgaaaatgta attcttgagt taacacttct
     5101 gggtggagaa tagggttgtt ttccccccac ataattggaa ggggaaggaa tatcatttaa
     5161 agctatggga gggtttcttt gattacaaca ctggagagaa atgcagcatg ttgctgattg
     5221 cctgtcacta aaacaggcca aaaactgagt ccttgggttg catagaaagc ttcatgttgc
     5281 taaaccaatg ttaagtgaat ctttggaaac aaaatgtttc caaattactg ggatgtgcat
     5341 gttgaaacgt gggttaatta actagccatg accaaaatcc cttaacgtga gttttcgttc
     5401 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5461 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5521 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5581 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5641 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5701 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5761 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5821 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5881 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5941 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6001 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6061 ctggcctttt gctggccttt tgctcacatg ttcttaatta aatttttcaa aagtagttga
     6121 caattaatca tcggcatagt atatcggcat agtataatac gactcactat aggagggcca
     6181 tcatggccaa gttgaccagt gctgtcccag tgctcacagc cagggatgtg gctggagctg
     6241 ttgagttctg gactgacagg ttggggttct ccagagattt tgtggaggat gactttgcag
     6301 gtgtggtcag agatgatgtc accctgttca tctcagcagt ccaggaccag gtggtgcctg
     6361 acaacaccct ggcttgggtg tgggtgagag gactggatga gctgtatgct gagtggagtg
     6421 aggtggtctc caccaacttc agggatgcca gtggccctgc catgacagag attggagagc
     6481 agccctgggg gagagagttt gccctgagag acccagcagg caactgtgtg cactttgtgg
     6541 cagaggagca ggactgagga taagaattgt aacaaaaaac cccgccccgg cggggttttt
     6601 tgttaattaa
//
