LOCUS       sequence-11488-e        6502 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 6502)
  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 6502)
  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: p6VP1
FEATURES             Location/Qualifiers
     source          1..6502
                     /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(3875..3899)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    3961..4549
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4014..4034)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4432..4443)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4591..5163
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
     rep_origin      5334..5922
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5823..5842
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        6008..6055
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        6047..6065
                     /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 accatgccct gccatcgcaa aggcaacggc ccaatccaga agctgccacg cgtgatcaag
     2761 aagggaggcg tcgaagtcat ggagaccgtc ccgttgtccg aggataccat ctacaaggtc
     2821 gaagccatcc tgctgcccaa cttcgccagc ggcagtaaca ccgccgtgta tcaaagcaga
     2881 ggcgcaccat acacattcac cgataccctc gacgctggca gcagcctctg ctacacactc
     2941 gccgtcgtca acctgccaga gatccccgaa gcactctgcg acgacaccct gctggtctgg
     3001 gaagcattcc gcgtcgagac cgagctcatc ttcacaccgc aagtcggatc cgctggctac
     3061 atccgcgcac aaggcacacc agctggagtg gaaggcagtc agatgtactt ctgggcatgc
     3121 ggagggtccc cactcgacgt gatcggcatc aatcccgacc cagagcgcat gaacgttgct
     3181 gccgggctcg aaggcccaag caaggagaac cagcccagcg tcgctggcat caaggccaca
     3241 cgcaagcagg tcaccgctgc aaacttccca atcgagatct ggtccgccga ccctacacgc
     3301 aacgagaact gccgctactt cggacgcatc gtcggcggat ccgtcactcc acctgtcgtg
     3361 tccttcggca accaatccac aaccccactg gtcgacgaga acggcgtcgg catcttgtgc
     3421 ctgttcggcg ccatctacct gaccagcgcc gacatgctcg gcatggtcgg ctacgctggc
     3481 aatccaacct tgtccgacgc atacagccag cagcgctccg tccaagccgc attcggccgc
     3541 ttcttccgcg tccatttccg ccagcgccgc gtcaagcacc cctacaccgt cgacatgatg
     3601 ttccgccagt tcctgcaacc ccagaagcct caggtccagg gaacccagcc aaacgctgtg
     3661 caggaagtgg tcatggagca gatgcagcca tctatcttgc ccactaccct cgaaggagcc
     3721 atcggctata gcccatccac caagttcatc ctgcaaaacg gagagctgat ctatccaagc
     3781 tccaccgtgg cagccggagc cgctaacctg ttcggaccgc ctgtggaaaa gcagacctcc
     3841 aaggagccca gcaagggcga actctgagac tagtacccag ctttcttgta caaagtggtt
     3901 cgatctagaa tggctagtgg atcccccggg ctgcaggaat tcgatatcaa gcttatcgat
     3961 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     4021 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     4081 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     4141 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     4201 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     4261 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     4321 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     4381 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     4441 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     4501 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccgt
     4561 cggcccactg ctccctaaac ctgagctagc attatcccta atacctgcca ccccactctt
     4621 aatcagtggt ggaagaacgg tctcagaact gtttgtttca attggccatt taagtttagt
     4681 agtaaaagac tggttaatga taacaatgca tcgtaaaacc ttcagaagga aaggagaatg
     4741 ttttgtggac cactttggtt ttcttttttg cgtgtggcag ttttaagtta ttagttttta
     4801 aaatcagtac tttttaatgg aaacaacttg accaaaaatt tgtcacagaa ttttgagacc
     4861 cattaaaaaa gttaaatgag aaacctgtgt gttcctttgg tcaacaccga gacatttagg
     4921 tgaaagacat ctaattctgg ttttacgaat ctggaaactt cttgaaaatg taattcttga
     4981 gttaacactt ctgggtggag aatagggttg ttttcccccc acataattgg aaggggaagg
     5041 aatatcattt aaagctatgg gagggtttct ttgattacaa cactggagag aaatgcagca
     5101 tgttgctgat tgcctgtcac taaaacaggc caaaaactga gtccttgggt tgcatagaaa
     5161 gcttcatgtt gctaaaccaa tgttaagtga atctttggaa acaaaatgtt tccaaattac
     5221 tgggatgtgc atgttgaaac gtgggttaat taactagcca tgaccaaaat cccttaacgt
     5281 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5341 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5401 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5461 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5521 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5581 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5641 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5701 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5761 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5821 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5881 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5941 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttcttaat taaatttttc
     6001 aaaagtagtt gacaattaat catcggcata gtatatcggc atagtataat acgactcact
     6061 ataggagggc catcatggcc aagttgacca gtgctgtccc agtgctcaca gccagggatg
     6121 tggctggagc tgttgagttc tggactgaca ggttggggtt ctccagagat tttgtggagg
     6181 atgactttgc aggtgtggtc agagatgatg tcaccctgtt catctcagca gtccaggacc
     6241 aggtggtgcc tgacaacacc ctggcttggg tgtgggtgag aggactggat gagctgtatg
     6301 ctgagtggag tgaggtggtc tccaccaact tcagggatgc cagtggccct gccatgacag
     6361 agattggaga gcagccctgg gggagagagt ttgccctgag agacccagca ggcaactgtg
     6421 tgcactttgt ggcagaggag caggactgag gataagaatt gtaacaaaaa accccgcccc
     6481 ggcggggttt tttgttaatt aa
//
