LOCUS       sequence-10345-e        5608 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 5608)
  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 5608)
  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: phGf
FEATURES             Location/Qualifiers
     source          1..5608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(8..28)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        13..342
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      193..328
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      367..376
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(373..375,376..378,379..1092)
                     /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(373..375,376..378,379..1089)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(418..439)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(679..698)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1026..1047
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1117..1238
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1128..1147)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1182..1201
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1439..1650
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             1663..1931
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     protein_bind    1973..2097
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2122..2127,2128..2145,2146..2152)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2206..2865
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2272..2291)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3207..3512
                     /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     3391..3410
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3553..3677)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    3697..4269
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
     rep_origin      4440..5028
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4929..4948
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5114..5161
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        5153..5171
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cccctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
       61 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
      121 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
      181 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
      241 tgactaattt tttttattta tacagaggcc gaggccgcct cggcctctga gctattccag
      301 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttga ttgggatcca
      361 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      421 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      481 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      541 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      601 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      661 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      721 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      781 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
      841 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
      901 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
      961 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1021 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1081 ctgtacaagt aaagcggccg cttcgagcag acatgataag atacattgat gagtttggac
     1141 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     1201 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     1261 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
     1321 aatgtggtaa aatcgataag gatccgggct ggcgtaatag cgaagaggcc cgcaccgatc
     1381 gcccttccca acagttgcgg tggagaagag catgcgtgag gctccggtgc ccgtcagtgg
     1441 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc
     1501 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
     1561 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
     1621 tttcgcaacg ggtttgccgc cagaacacag ctgaagcttc gaggggctcg catctctcct
     1681 tcacgcgccc gccgccctac ctgaggccgc catccacgcc ggttgagtcg cgttctgccg
     1741 cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt aaagctcagg
     1801 tcgagaccgg gcctttgtcc ggcgctccct tggagcctac ctagactcag ccggctctcc
     1861 acgctttgcc tgaccctgct tgctcaactc tacgtctttg tttcgttttc tgttctgcgc
     1921 cgttacagat ccaagctgtg accggcgcct acgtaagtga tagcttgatc aaacaagttt
     1981 gtacaaaaaa gctgaacgag aaacgtaaaa tgatataaat atcaatatat taaattagat
     2041 tttgcataaa aaacagacta cataatactg taaaacacaa catatccagt cactatggcg
     2101 gccgcattag gcaccccagg ctttacactt tatgcttccg gctcgtataa tgtgtggatt
     2161 ttgagttagg atccgtcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc
     2221 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
     2281 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta
     2341 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc
     2401 ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct ggtgatatgg
     2461 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc
     2521 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg
     2581 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc
     2641 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac
     2701 ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg
     2761 ccgctggcga ttcaggttca tcatgccgtc tgtgatggct tccatgtcgg cagaatgctt
     2821 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaatctag aggatccggc
     2881 ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg atttttgcgg tataagaata
     2941 tatactgata tgtatacccg aagtatgtca aaaagaggtg tgctatgaag cagcgtatta
     3001 cagtgacagt tgacagcgac agctatcagt tgctcaaggc atatatgatg tcaatatctc
     3061 cggtctggta agcacaacca tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa
     3121 agcggaaaat caggaaggga tggctgaggt cgcccggttt attgaaatga acggctcttt
     3181 tgctgacgag aacagggact ggtgaaatgc agtttaaggt ttacacctat aaaagagaga
     3241 gccgttatcg tctgtttgtg gatgtacaga gtgatattat tgacacgccc gggcgacgga
     3301 tggtgatccc cctggccagt gcacgtctgc tgtcagataa agtctcccgt gaactttacc
     3361 cggtggtgca tatcggggat gaaagctggc gcatgatgac caccgatatg gccagtgtgc
     3421 cggtctccgt tatcggggaa gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa
     3481 acgccattaa cctgatgttc tggggaatat aaatgtcagg ctccgttata cacagccagt
     3541 ctgcaggtcg accatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt
     3601 ttatgcaaaa tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcagct
     3661 ttcttgtaca aagtggttcg atctagaatg gctagcatta tccctaatac ctgccacccc
     3721 actcttaatc agtggtggaa gaacggtctc agaactgttt gtttcaattg gccatttaag
     3781 tttagtagta aaagactggt taatgataac aatgcatcgt aaaaccttca gaaggaaagg
     3841 agaatgtttt gtggaccact ttggttttct tttttgcgtg tggcagtttt aagttattag
     3901 tttttaaaat cagtactttt taatggaaac aacttgacca aaaatttgtc acagaatttt
     3961 gagacccatt aaaaaagtta aatgagaaac ctgtgtgttc ctttggtcaa caccgagaca
     4021 tttaggtgaa agacatctaa ttctggtttt acgaatctgg aaacttcttg aaaatgtaat
     4081 tcttgagtta acacttctgg gtggagaata gggttgtttt ccccccacat aattggaagg
     4141 ggaaggaata tcatttaaag ctatgggagg gtttctttga ttacaacact ggagagaaat
     4201 gcagcatgtt gctgattgcc tgtcactaaa acaggccaaa aactgagtcc ttgggttgca
     4261 tagaaagctt catgttgcta aaccaatgtt aagtgaatct ttggaaacaa aatgtttcca
     4321 aattactggg atgtgcatgt tgaaacgtgg gttaattaac tagccatgac caaaatccct
     4381 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4441 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4501 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4561 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     4621 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4681 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4741 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4801 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4861 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4921 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4981 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5041 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt cttaattaaa
     5101 tttttcaaaa gtagttgaca attaatcatc ggcatagtat atcggcatag tataatacga
     5161 ctcactatag gagggccatc atggccaagt tgaccagtgc tgtcccagtg ctcacagcca
     5221 gggatgtggc tggagctgtt gagttctgga ctgacaggtt ggggttctcc agagattttg
     5281 tggaggatga ctttgcaggt gtggtcagag atgatgtcac cctgttcatc tcagcagtcc
     5341 aggaccaggt ggtgcctgac aacaccctgg cttgggtgtg ggtgagagga ctggatgagc
     5401 tgtatgctga gtggagtgag gtggtctcca ccaacttcag ggatgccagt ggccctgcca
     5461 tgacagagat tggagagcag ccctggggga gagagtttgc cctgagagac ccagcaggca
     5521 actgtgtgca ctttgtggca gaggagcagg actgaggata agaattgtaa caaaaaaccc
     5581 cgccccggcg gggttttttg ttaattaa
//
