LOCUS       sequence-222810-        7549 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of MCPyV LT mutant (Ser 220 to Ala).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7549)
  AUTHORS   Kwun HJ, Chang Y, Moore PS
  TITLE     Protein-mediated viral latency is a novel mechanism for Merkel cell 
            polyomavirus persistence.
  JOURNAL   Proc Natl Acad Sci U S A. 2017 May 16;114(20):E4040-E4047. doi: 
            10.1073/pnas.1703879114. Epub 2017 May 1.
  PUBMED    28461484
REFERENCE   2  (bases 1 to 7549)
  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"; doi: 
            "10.1073/pnas.1703879114"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2017-05-16- 16"; volume: "114"; issue: "20"; pages: 
            "E4040-E4047"
COMMENT     Sequence Label: pcDNA6 MCV LTco S220A
FEATURES             Location/Qualifiers
     source          1..7549
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3505..3522)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3511..3735
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3781..4209
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3868..3887)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4078..4099
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4218..4238)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4223..4552
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4403..4538
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4600..4647
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4666..5064
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5222..5343
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5259..5278)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5313..5332
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5392..5408)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5392..5408)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5405..5427)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5416..5432
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5440..5446,5447..5464,5465..5470))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5485..5506
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5623..5640)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5794..6382)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(6553..7344,7345..7413))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7176..7195
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7414..7518)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gggtaccgca gatatccacc atggatctgg tactgaatcg caaagaaagg gaggcgttgt
      961 gtaaactcct tgagattgct cccaattgtt atgggaacat tcccctgatg aaggcagcgt
     1021 ttaagcgatc ctgtttgaaa caccatccgg ataaaggcgg aaacccagtc attatgatgg
     1081 agctgaatac cctctggtcg aagtttcagc agaatatcca taagttgaga tcggatttct
     1141 cgatgtttga cgaggtggac gaggccccga tctacggaac aacaaagttc aaagaatggt
     1201 ggcgctcagg gggattttcg tttggtaaag cctatgagta tggacccaac ccccatggga
     1261 cgaactcccg gagccgaaaa ccgtcctcga atgcgtcacg gggtgcaccg tcagggtcgt
     1321 cccctcccca ttcgcagtcg agctcgtcgg ggtacgggag cttctcagcg tcgcaggcca
     1381 gcgatagcca gtcgcgggga cctgatattc cccctgaaca ccatgaggaa ccgacgtcgt
     1441 caagcggatc gtccagccga gaagaaacta ccaatagcgg cagggaatca tcaacgccca
     1501 acgggacgtc cgtgccgagg aactcctcaa gaacggacgg aacttgggaa gatcttttct
     1561 gcgacgaatc gctgtcagcc cccgaacccc cgtcgtcatc cgaggaaccc gaagaaccgc
     1621 cttcgtcaag atcatcaccc cgccagcccc catcatcaag cgcggaggaa gcatcgtcgt
     1681 cccagtttac agacgaggag taccgcagct catcgtttac aactccaaaa acacctccgc
     1741 ccttttcccg gaagagaaag ttcgggggat cgcgctcctc ggcctcctcc gcgtcctccg
     1801 catcgttcac gtcaacgcca ccgaagccga aaaagaatag ggagacacca gtgcccacag
     1861 actttcctat tgatctgagc gattacttgt cgcacgcggt ctattcaaac aagacggtca
     1921 gctgcttcgc gatctacacc acaagcgaca aagccattga gctctatgat aagattgaaa
     1981 agttcaaggt ggatttcaaa tcgagacatg cgtgtgaact gggctgcatc ttgctgttca
     2041 tcacgctgtc caaacaccga gtctcagcta tcaaaaactt ttgctcaacc ttctgcacca
     2101 tctcgttctt gatttgcaaa ggagtcaata agatgccaga gatgtataac aacttgtgca
     2161 agcctcccta caagctcctc caagagaaca agccgctttt gaattacgaa ttccaagaaa
     2221 aggagaaaga ggcgtcatgc aattggaatc ttgtcgcgga gttcgcatgc gaatatgaac
     2281 ttgacgacca tttcatcatt cttgcgcact acttggactt tgcgaagcca ttcccgtgcc
     2341 aaaagtgcga gaacagatcg agacttaaac cacataaggc ccacgaggcc caccattcca
     2401 acgccaagct cttttacgaa tccaaatcgc aaaagaccat ctgtcagcag gctgccgaca
     2461 cggtcctggc gaaacgaagg cttgagatgc tcgaaatgac tcgcacggaa atgttgtgta
     2521 agaagttcaa aaagcacctc gaacggctgc gggatttgga cacaatcgac ctcctctatt
     2581 acatgggtgg ggtagcttgg tactgctgcc tttttgaaga gtttgaaaag aagctgcaga
     2641 agattatcca gctccttact gagaacattc caaagtatag aaacatttgg ttcaagggac
     2701 ccatcaattc gggaaaaacc agcttcgcag ctgcactgat cgacctgctc gaaggaaaag
     2761 ccctgaacat taactgtcct tcggacaaac ttcccttcga gcttggttgt gcattggaca
     2821 agtttatggt agtgtttgag gatgtgaagg gacagaattc actgaataag gatcttcagc
     2881 ctggtcaggg tatcaacaat ttggacaatc ttcgggacca cttggacgga gcggtggccg
     2941 tgtccttgga gaaaaagcac gtgaataaga agcaccagat ctttccccca tgcatcgtaa
     3001 cggctaatga ttacttcatc cccaagacac ttattgcgcg cttttcgtat acactccatt
     3061 tctcacccaa agcgaatctc cgcgactcat tggatcagaa catggaaatc agaaaaaggc
     3121 ggattctcca gtcgggcacg acccttcttc tgtgtctcat ctggtgcctt cccgatacta
     3181 ctttcaagcc ttgtctccaa gaagagatca agaattggaa acaaatcctc cagagcgaga
     3241 tcagctatgg caaattctgt cagatgattg agaacgtaga agcgggtcag gacccgttgt
     3301 tgaacatcct catcgaggag gagggtccgg aagagacaga ggaaacccaa gattcgggga
     3361 cgttctcgca atagccggga ctggaggtca acaaaccggc ggatcaaccg aggctctgtc
     3421 tgttggtggc gagccacctt ctgttcgcac ctcccccgtg tttgccttga cccgggctcg
     3481 agtccgcgga aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     3541 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     3601 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     3661 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     3721 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     3781 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3841 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3901 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3961 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     4021 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4081 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     4141 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4201 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4261 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4321 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4381 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4441 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     4501 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     4561 gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca
     4621 tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc caagcctttg
     4681 tctcaagaag aatccaccct cattgaaaga gcaacggcta caatcaacag catccccatc
     4741 tctgaagact acagcgtcgc cagcgcagct ctctctagcg acggccgcat cttcactggt
     4801 gtcaatgtat atcattttac tgggggacct tgtgcagaac tcgtggtgct gggcactgct
     4861 gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga tcggaaatga gaacaggggc
     4921 atcttgagcc cctgcggacg gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa
     4981 gccatagtga aggacagtga tggacagccg acggcagttg ggattcgtga attgctgccc
     5041 tctggttatg tgtgggaggg ctaagcactt cgtggccgag gagcaggact gacacgtgct
     5101 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5161 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5221 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5281 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5341 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     5401 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5461 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5521 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5581 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5641 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5701 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5761 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5821 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5881 ccaggcgttt gcccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5941 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6001 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6061 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6121 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6181 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6241 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6301 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6361 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6421 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6481 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6541 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6601 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6661 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6721 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6781 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6841 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6901 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6961 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7021 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7081 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7141 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7201 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7261 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7321 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7381 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7441 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7501 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
