LOCUS       sequence-173662-        7006 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Flag/HA tagged MmuPV1 E6 R130A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7006)
  AUTHORS   Meyers JM, Uberoi A, Grace M, Lambert PF, Munger K
  TITLE     Cutaneous HPV8 and MmuPV1 E6 Proteins Target the NOTCH and TGF-beta 
            Tumor Suppressors to Inhibit Differentiation and Sustain 
            Keratinocyte Proliferation.
  JOURNAL   PLoS Pathog. 2017 Jan 20;13(1):e1006171. doi: 
            10.1371/journal.ppat.1006171.
  PUBMED    28107544
REFERENCE   2  (bases 1 to 7006)
  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."; date: "2017-01-20"; volume: "13(1):e1006171. doi"; pages: 
            " 10.1371/journal.ppat.1006171"
COMMENT     Sequence Label: pNCMV MmuPV1 E6 R130A
FEATURES             Location/Qualifiers
     source          1..7006
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..24
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             37..63
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(591..610)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    656..711
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(710..729)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1069..1085)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1069..1085)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1082..1104)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1093..1109
                     /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(1117..1123,1124..1141,1142..1147))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1162..1183
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1300..1317)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1471..2059)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1551..1570)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2230..3021,3022..3090))
                     /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     2853..2872
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3091..3195)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3263..3281
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3319..3341)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3441..3460
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3654..3676
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3668..3685
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3669..3685
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4096..4241
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             4298..5092
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4352..4371)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4962..4981
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5286..5305)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5330..5377
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     enhancer        5670..6049
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6050..6253
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6204..6224
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6250..6274
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          6365..6937
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
ORIGIN
        1 gactacaagg atgacgatga caagctcgat ggaggatacc catacgatgt tccagattac
       61 gctgatatcg gatccgaaat cggcaaaggc tacactctcg aggaggtgct tagatattct
      121 aacaaagatg tcgtggattt tcatttgtct tgtgcttttt gctctactac tatggatcat
      181 aacgagaagg ccagattcat acaggctaaa ttgaaatgtg ttgttagaga ttttgctttt
      241 aaaggtgctt gtattgtgtg ccgcagacag cttgcttgca aggaaaagct tttgcatact
      301 agagttacag gggaggctga tttggtagag tgcatggctg gcaagaatat tgtgtttgtt
      361 actgtaagat gtgttacgtg cctggcactc cttactgcct ctgaaaagct tgatgccaaa
      421 gcgtgcggct tgccatttca cttggtgcgc cacatgtggg caggctactg cgggttctgc
      481 aaaccattac tataaggatc cgtcgaggaa ttcactcctc aggtgcaggc tgcctatcag
      541 aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc
      601 tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa
      661 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg
      721 acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca
      781 acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat
      841 atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag
      901 atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta
      961 catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct
     1021 cttctcttat ggagatccct cgacggatcg gccgcaattc gtaatcatgt catagctgtt
     1081 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     1141 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     1201 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     1261 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     1321 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     1381 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     1441 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     1501 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     1561 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     1621 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     1681 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     1741 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     1801 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     1861 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     1921 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     1981 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     2041 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     2101 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     2161 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     2221 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     2281 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     2341 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     2401 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     2461 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     2521 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     2581 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     2641 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     2701 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     2761 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     2821 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     2881 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     2941 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     3001 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     3061 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     3121 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     3181 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     3241 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     3301 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     3361 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     3421 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     3481 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     3541 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     3601 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     3661 acgacgttgt aaaacgacgg ccagtgccaa gcttcacgct gccgcaagca ctcagggcgc
     3721 aagggctgct aaaggaagcg gaacacgtag aaagccagtc cgcagaaacg gtgctgaccc
     3781 cggatgaatg tcagctactg ggctatctgg acaagggaaa acgcaagcgc aaagagaaag
     3841 caggtagctt gcagtgggct tacatggcga tagctagact gggcggtttt atggacagca
     3901 agcgaaccgg aattgccagc tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta
     3961 aactggatgg ctttcttgcc gccaaggatc tgatggcgca ggggatcaag atcctgcttc
     4021 atccccgtgg cccgttgctc gcgtttgctg gcggtgtccc cggaagaaat atatttgcat
     4081 gtctttagtt ctatgatgac acaaaccccg cccagcgtct tgtcattggc gaattcgaac
     4141 acgcagatgc agtcggggcg gcgcggtccc aggtccactt cgcatattaa ggtgacgcgt
     4201 gtggcctcga acaccgagcg accctgcagc gacccgctta acagcgtcaa cagcgtgccg
     4261 cagatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4321 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4381 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4441 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4501 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4561 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4621 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4681 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4741 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4801 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     4861 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4921 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4981 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5041 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5101 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     5161 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5221 atcctccagc gcggggatct catgctggag ttcttcgccc accccgggag atgggggagg
     5281 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     5341 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     5401 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     5461 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     5521 ggggcggcaa gccctgccat agccacgggc cccgtgggtt agggacggcg gatcgcggcc
     5581 ctctagagag cttggcccat tgcatacgtt gtatccatat cataatatgt acatttatat
     5641 tggctcatgt ccaacattac cgccatgttg acattgatta ttgactagtt attaatagta
     5701 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     5761 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     5821 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     5881 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     5941 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     6001 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     6061 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     6121 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     6181 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     6241 tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt
     6301 tgacctccat agaagacacc gggaccgatc cagcctccgg tcgaccgatc ctgagaactt
     6361 cagggtgagt ttggggaccc ttgattgttc tttctttttc gctattgtaa aattcatgtt
     6421 atatggaggg ggcaaagttt tcagggtgtt gtttagaatg ggaagatgtc ccttgtatca
     6481 ccatggaccc tcatgataat tttgtttctt tcactttcta ctctgttgac aaccattgtc
     6541 tcctcttatt ttcttttcat tttctgtaac tttttcgtta aactttagct tgcatttgta
     6601 acgaattttt aaattcactt ttgtttattt gtcagattgt aagtactttc tctaatcact
     6661 tttttttcaa ggcaatcagg gtatattata ttgtacttca gcacagtttt agagaacaat
     6721 tgttataatt aaatgataag gtagaatatt tctgcatata aattctggct ggcgtggaaa
     6781 tattcttatt ggtagaaaca actacaccct ggtcatcatc ctgcctttct ctttatggtt
     6841 acaatgatat acactgtttg agatgaggat aaaatactct gagtccaaac cgggcccctc
     6901 tgctaaccat gttcatgcct tcttctcttt cctacagctc ctgggcaacg tgctggttgt
     6961 tgtgctgtct catcattttg gcaaagaatt cctcgacgga tctatg
//
