LOCUS       sequence-10348-e        6493 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 6493)
  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 6493)
  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: pwP
FEATURES             Location/Qualifiers
     source          1..6493
                     /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(3866..3890)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    3952..4540
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4005..4025)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4423..4434)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4582..5154
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
     rep_origin      5325..5913
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5814..5833
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5999..6046
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        6038..6056
                     /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 ttccggaacc
     2701 accatggcac caaagcgcaa gtcaggggtc agcaaatgtg aaaccaagtg caccaaggca
     2761 tgcccccgcc ctgcacccgt cccaaagctc ttgatcaagg gcggcatgga agtcctcgat
     2821 ctggtcaccg gccccgattc cgtcaccgag atcgaggcat tcctgaatcc acgcatgggc
     2881 caacccccaa cacccgagtc actgaccgaa ggcggccagt attacggctg gagccgcggc
     2941 atcaacctcg ccaccagcga caccgaagac agtcccgaga acaacacctt gccaacctgg
     3001 tccatggcca aactgcaact gccaatgctg aacgaagatc tgacatgcga tacattgcaa
     3061 atgtgggaag ccgtcagcgt caagacagaa gtcgtcggga gcgggagcct gctcgatgtc
     3121 cacggcttca acaagccaac cgacaccgtg aacaccaagg gcatcagtac cccggttgag
     3181 gggtcacagt accacgtctt cgccgtcggg ggcgagcccc tggatctgca aggcttggtc
     3241 accgacgcac gcaccaagtt caaagaggag ggcgtggtca ccatcaagac cattaccaag
     3301 aaggacatgg tcaacaagga tcaggtgctc aaccccatct caaaagcaaa gctcgacaag
     3361 gatggcatgt accccgtcga gatttggcac cccgaccccg ccaagaacga aaacacccgc
     3421 tacttcggga actataccgg cgggaccacc accccaccgg tgctccaatt caccaataca
     3481 ctcaccaccg tcctgctgga cgagaacggc gtcggcccac tgtgcaaggg cgaagggttg
     3541 tatctgagtt gcgtggacat catggggtgg cgcgtcaccc gcaactacga cgcacaccat
     3601 tggcgcggcc tgccacgcta cttcaagatt acactccgca agcgctgggt caagaaccca
     3661 tacccaatgg caagtctgat ctcgagtctg ttcaacaata tgttgccaca agtccagggg
     3721 cagccaatgg agggcgaaaa cacacaagtg gaagaagtcc gcgtctacga cggcaccgag
     3781 cccgtgcccg gcgatcccga catgaccaga tacgtcgata ggttcggcaa gaccaaaacc
     3841 gtgttcccag gaaactgagc ctaggaccca gctttcttgt acaaagtggt tcgatctaga
     3901 atggctagtg gatcccccgg gctgcaggaa ttcgatatca agcttatcga taatcaacct
     3961 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     4021 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     4081 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     4141 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     4201 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     4261 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     4321 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     4381 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     4441 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     4501 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg tcggcccact
     4561 gctccctaaa cctgagctag cattatccct aatacctgcc accccactct taatcagtgg
     4621 tggaagaacg gtctcagaac tgtttgtttc aattggccat ttaagtttag tagtaaaaga
     4681 ctggttaatg ataacaatgc atcgtaaaac cttcagaagg aaaggagaat gttttgtgga
     4741 ccactttggt tttctttttt gcgtgtggca gttttaagtt attagttttt aaaatcagta
     4801 ctttttaatg gaaacaactt gaccaaaaat ttgtcacaga attttgagac ccattaaaaa
     4861 agttaaatga gaaacctgtg tgttcctttg gtcaacaccg agacatttag gtgaaagaca
     4921 tctaattctg gttttacgaa tctggaaact tcttgaaaat gtaattcttg agttaacact
     4981 tctgggtgga gaatagggtt gttttccccc cacataattg gaaggggaag gaatatcatt
     5041 taaagctatg ggagggtttc tttgattaca acactggaga gaaatgcagc atgttgctga
     5101 ttgcctgtca ctaaaacagg ccaaaaactg agtccttggg ttgcatagaa agcttcatgt
     5161 tgctaaacca atgttaagtg aatctttgga aacaaaatgt ttccaaatta ctgggatgtg
     5221 catgttgaaa cgtgggttaa ttaactagcc atgaccaaaa tcccttaacg tgagttttcg
     5281 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5341 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5401 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5461 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5521 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5581 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5641 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5701 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5761 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5821 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5881 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5941 ttcctggcct tttgctggcc ttttgctcac atgttcttaa ttaaattttt caaaagtagt
     6001 tgacaattaa tcatcggcat agtatatcgg catagtataa tacgactcac tataggaggg
     6061 ccatcatggc caagttgacc agtgctgtcc cagtgctcac agccagggat gtggctggag
     6121 ctgttgagtt ctggactgac aggttggggt tctccagaga ttttgtggag gatgactttg
     6181 caggtgtggt cagagatgat gtcaccctgt tcatctcagc agtccaggac caggtggtgc
     6241 ctgacaacac cctggcttgg gtgtgggtga gaggactgga tgagctgtat gctgagtgga
     6301 gtgaggtggt ctccaccaac ttcagggatg ccagtggccc tgccatgaca gagattggag
     6361 agcagccctg ggggagagag tttgccctga gagacccagc aggcaactgt gtgcactttg
     6421 tggcagagga gcaggactga ggataagaat tgtaacaaaa aaccccgccc cggcggggtt
     6481 ttttgttaat taa
//
