LOCUS       sequence-422376-        6971 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 6971)
  AUTHORS   Shuda M, Feng H, Kwun HJ, Rosen ST, Gjoerup O, Moore PS, Chang Y
  TITLE     T antigen mutations are a human tumor-specific signature for Merkel 
            cell polyomavirus.
  JOURNAL   Proc Natl Acad Sci U S A. 2008 Oct 21. 105(42):16272-7.
  PUBMED    18812503
REFERENCE   2  (bases 1 to 6971)
  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: "Proc Natl 
            Acad Sci U S A. 2008 Oct 21. 105(42):16272-7."
COMMENT     Sequence Label: pcDNA6.MCV.LT339.V5.4
FEATURES             Location/Qualifiers
     source          1..6971
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        121..500
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        501..704
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     655..675
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     749..768
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        749..767
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2722..2763
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2773..2790
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2813..2830)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2819..3043
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3089..3517
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3176..3195)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3386..3407
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3526..3546)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3531..3860
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3711..3846
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3773..3792
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3908..3955
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3974..4372
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4530..4651
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4567..4586)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4621..4640
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4700..4716)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4700..4716)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4713..4735)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4724..4740
                     /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(4748..4754,4755..4772,4773..4778))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4793..4814
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4931..4948)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5102..5690)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5182..5201)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5861..6652,6653..6721))
                     /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     6484..6503
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6722..6826)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6901..6920)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
       61 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
      121 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
      181 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      241 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      301 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      361 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      421 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      481 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      541 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      601 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      661 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      721 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      781 gctagcgttt aaacttaagc ttggtaccga gctcggatcc actagtccag tgtggtggaa
      841 ttctgcagat atcatggatt tagtcctaaa caggaaagaa agagaggctc tctgcaagct
      901 tttagagatt gctcctaatt gttatggcaa catccctctg atgaaagctg ctttcaaaag
      961 aagctgctta aagcatcacc ctgataaagg gggaaatcct gttataatga tggaattgaa
     1021 caccctttgg agcaaattcc agcaaaatat ccacaagctc agaagtgact tctctatgtt
     1081 tgatgaggtc agtacaaaat ttccttggga agaatatgga actttaaagg attatatgca
     1141 aagtggatat aatgctagat tttgcagagg tcctgggtgc atgcttaagc aacttagaga
     1201 ttctaagtgc gcttgtatta gctgtaagtt gtctcgccag cattgtagtc taaaaacttt
     1261 aaagcaaaaa aactgtctga cgtggggaga gtgtttttgc tatcagtgct ttattctttg
     1321 gtttggattt cctcctactt gggaaagttt tgactggtgg caaaaaactt tagaagaaac
     1381 tgactactgc ttactgcatc tgcacctttt ctagactcct acttccttcc tctgtaagta
     1441 ttagatatgg aaaagtctat aaggcaaaat atcaaagaaa ggttatttat gacagatttt
     1501 ctgtactttc ccatctaggt tgacgaggcc cctatatatg ggaccactaa attcaaagaa
     1561 tggtggagat caggaggatt cagcttcggg aaggcatacg aatatgggcc caatccacac
     1621 gggaccaact caagatccag aaagccttcc tccaatgcat ccaggggagc ccccagtgga
     1681 agctcaccac cccacagcca gagctcttcc tctgggtatg ggtccttctc agcgtcccag
     1741 gcttcagact cccagtccag aggacccgat atacctcccg aacaccatga ggaacccacc
     1801 tcatcctctg gatccagtag cagagaggag accaccaatt caggaagaga atccagcaca
     1861 ccaaatggaa ccagtgtacc tagaaattct tccagaactg atggcacctg ggaggatctc
     1921 ttctgcgatg aatcactttc ctcccctgag cctccctcgt cctctgagga gcctgaggag
     1981 cccccctcct caagaagctc gccccggcag cccccgtctt cctctgccga ggaggcctcg
     2041 tcatctcagt ttacagatga ggaatacaga tcctcctcct tcaccacccc gaagacccct
     2101 cctccattct caagaaagcg aaaatttggg gggtcccgaa gctctgcaag ctctgctagt
     2161 tcagcaagtt ttacaagcac tccaccaaag ccaaaaaaga acagagaaac tcctgttcct
     2221 actgattttc ctattgatct ttctgattat cttagccatg ctgtatatag taataaaaca
     2281 gtaagttgtt ttgccattta tactacttct gataaagcta tagagttata tgataagatt
     2341 gagaaattta aagttgattt taaaggcagg catgcctgtg aattaggatg tattttattg
     2401 tttataactt tatcaaagca tagagtatct gctattaaga atttctgctc taccttctgc
     2461 actataagct ttttaatttg taaaggagtg aataagatgc ctgaaatgta taataattta
     2521 tgcaagcccc cttacaaatt actgcaagag aataagccac tgctcaatta tgaatttcaa
     2581 gaaaaagaaa aagaggccag ctgcaattgg aatttagttg ctgaatttgc ttgtgaatat
     2641 gagctagacg aggttagaga tgctggaaat gaccaggaca gaaatgctat gtactcgagt
     2701 ctagagggcc cgcggttcga aggtaagcct atccctaacc ctctcctcgg tctcgattct
     2761 acgcgtaccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc agcctcgact
     2821 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     2881 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     2941 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     3001 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     3061 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     3121 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3181 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3241 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3301 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3361 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3421 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3481 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     3541 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3601 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3661 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3721 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3781 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3841 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     3901 agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     3961 gaggaactaa accatggcca agcctttgtc tcaagaagaa tccaccctca ttgaaagagc
     4021 aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct
     4081 ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg
     4141 tgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat
     4201 cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt
     4261 gcttctcgat ctgcatcctg ggatcaaagc catagtgaag gacagtgatg gacagccgac
     4321 ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggct aagcacttcg
     4381 tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg
     4441 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4501 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     4561 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4621 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     4681 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     4741 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     4801 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     4861 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     4921 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4981 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5041 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5101 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5161 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5221 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5281 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5341 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5401 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5461 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5521 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     5581 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5641 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5701 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5761 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5821 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5881 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5941 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6001 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6061 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6121 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6181 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6241 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6301 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6361 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6421 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6481 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6541 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6601 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6661 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6721 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6781 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6841 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     6901 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     6961 ggtcgctgag t
//
