LOCUS       sequence-11489-e        6481 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 6481)
  AUTHORS   Schowalter RM, Pastrana DV, Pumphrey KA, Moyer AL, Buck CB.
  TITLE     Merkel cell polyomavirus and two previously unknown polyomaviruses 
            are chronically shed from human skin.
  JOURNAL   Cell Host Microbe. 2010 Jun 25;7(6):509-15.
  PUBMED    20542254
REFERENCE   2  (bases 1 to 6481)
  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: "Cell Host 
            Microbe."; date: "2010-06-25"; volume: "7(6)"; pages: "509-15"
COMMENT     Sequence Label: p7VP1
FEATURES             Location/Qualifiers
     source          1..6481
                     /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"
     protein_bind    complement(3854..3878)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    3940..4528
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3993..4013)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4411..4422)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4570..5142
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
     rep_origin      5313..5901
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5802..5821
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5987..6034
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        6026..6044
                     /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 accatgccat gccagcgcaa gggcaacggc ccaacccaga agctgccacg cgtcatccgc
     2761 aagggaggcg tcgaggtcct cgacaccgtg cccctgaccg aagataccca gtacaaggtc
     2821 gaggccgtcc tgctgcctaa cttcggcaag gcagccacta ccggaaactt ccaaagccgc
     2881 ggactgccct acaccatgtc cgataccctc ggccctggcg ctgccctgtg ctatagcgtc
     2941 gccgtcatca acctgcccga gatccccgac gcaatgtgcg aagataccat gatcgtctgg
     3001 gaagcatacc gcctcgagac cgagctgctg ttcgcacctc agatggcatc cagcggatac
     3061 cagcgcgcta acggaacact cgctggcatc gaaggatccc agctgtactt ctgggcatgc
     3121 ggaggaggtc cactcgacgt gatcggcatc aatcccgatc ccgagcgcct gaaagtgaac
     3181 gaggcactcg agggcccagg aaacaccgac gtggcgtccc tgcaagcact gcgcaagcag
     3241 gtcaacgctg ccaactttcc ggttgaactg tgggtcgccg acccaaccaa gaacgacaac
     3301 acacgctact tcggacgcgt cgtcggcgga ggcgtgacac cgcccgtcgt cagttacggc
     3361 aaccagtcca caaccccact catcgacgag aacggcgtcg gcatcctgtg caccttcggc
     3421 tccgtctacc tgacatccgc cgacatggtc ggcatgaccg ggctgccagg actcccaacc
     3481 ctgtccgccg attactccaa tcagcgcacc gtgcaggcag gctacggccg gttcttccgc
     3541 gtgcactgtc gccagcgccg catcaagcat ccatacaccg tcgacatgat gttccgccag
     3601 ttcctgcaac cacagaagcc tcaggtccag ggacagcagc ctgccgctgt gcaggaggtc
     3661 accatggaac agatgcaacc agcaaccatc ccgccaactg tcgaaggagg cctcggattc
     3721 gccccaacat ccaagttcct gattcagaac ggcgaactga tctacccatc cagcaacgca
     3781 gctgctgcgg ctgcaaagat cagcgtggca cccaagaaga ataccgacaa caagaaggag
     3841 ctctgagact agtacccagc tttcttgtac aaagtggttc gatctagaat ggctagtgga
     3901 tcccccgggc tgcaggaatt cgatatcaag cttatcgata atcaacctct ggattacaaa
     3961 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     4021 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     4081 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     4141 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     4201 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     4261 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     4321 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
     4381 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     4441 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     4501 cggatctccc tttgggccgc ctccccgcat cgataccgtc ggcccactgc tccctaaacc
     4561 tgagctagca ttatccctaa tacctgccac cccactctta atcagtggtg gaagaacggt
     4621 ctcagaactg tttgtttcaa ttggccattt aagtttagta gtaaaagact ggttaatgat
     4681 aacaatgcat cgtaaaacct tcagaaggaa aggagaatgt tttgtggacc actttggttt
     4741 tcttttttgc gtgtggcagt tttaagttat tagtttttaa aatcagtact ttttaatgga
     4801 aacaacttga ccaaaaattt gtcacagaat tttgagaccc attaaaaaag ttaaatgaga
     4861 aacctgtgtg ttcctttggt caacaccgag acatttaggt gaaagacatc taattctggt
     4921 tttacgaatc tggaaacttc ttgaaaatgt aattcttgag ttaacacttc tgggtggaga
     4981 atagggttgt tttcccccca cataattgga aggggaagga atatcattta aagctatggg
     5041 agggtttctt tgattacaac actggagaga aatgcagcat gttgctgatt gcctgtcact
     5101 aaaacaggcc aaaaactgag tccttgggtt gcatagaaag cttcatgttg ctaaaccaat
     5161 gttaagtgaa tctttggaaa caaaatgttt ccaaattact gggatgtgca tgttgaaacg
     5221 tgggttaatt aactagccat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5281 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5341 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5401 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5461 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5521 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     5581 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5641 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5701 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5761 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5821 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5881 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5941 tgctggcctt ttgctcacat gttcttaatt aaatttttca aaagtagttg acaattaatc
     6001 atcggcatag tatatcggca tagtataata cgactcacta taggagggcc atcatggcca
     6061 agttgaccag tgctgtccca gtgctcacag ccagggatgt ggctggagct gttgagttct
     6121 ggactgacag gttggggttc tccagagatt ttgtggagga tgactttgca ggtgtggtca
     6181 gagatgatgt caccctgttc atctcagcag tccaggacca ggtggtgcct gacaacaccc
     6241 tggcttgggt gtgggtgaga ggactggatg agctgtatgc tgagtggagt gaggtggtct
     6301 ccaccaactt cagggatgcc agtggccctg ccatgacaga gattggagag cagccctggg
     6361 ggagagagtt tgccctgaga gacccagcag gcaactgtgt gcactttgtg gcagaggagc
     6421 aggactgagg ataagaattg taacaaaaaa ccccgccccg gcggggtttt ttgttaatta
     6481 a
//
