LOCUS       sequence-180205-        7006 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Flag/HA tagged MmuPV1 E6.
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
FEATURES             Location/Qualifiers
     source          1..7006
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          696..1268
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     CDS             1338..1361
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1374..1400
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1928..1947)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1993..2048
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2047..2066)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2406..2422)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2406..2422)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2419..2441)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2430..2446
                     /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(2454..2460,2461..2478,2479..2484))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2499..2520
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2637..2654)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2808..3396)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2888..2907)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3567..4358,4359..4427))
                     /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     4190..4209
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4428..4532)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4600..4618
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4656..4678)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4778..4797
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4991..5013
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5005..5022
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5006..5022
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5433..5578
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             5635..6429
                     /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(5689..5708)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6299..6318
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6623..6642)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6667..6714
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg gtcgaccgat cctgagaact tcagggtgag tttggggacc cttgattgtt
      721 ctttcttttt cgctattgta aaattcatgt tatatggagg gggcaaagtt ttcagggtgt
      781 tgtttagaat gggaagatgt cccttgtatc accatggacc ctcatgataa ttttgtttct
      841 ttcactttct actctgttga caaccattgt ctcctcttat tttcttttca ttttctgtaa
      901 ctttttcgtt aaactttagc ttgcatttgt aacgaatttt taaattcact tttgtttatt
      961 tgtcagattg taagtacttt ctctaatcac ttttttttca aggcaatcag ggtatattat
     1021 attgtacttc agcacagttt tagagaacaa ttgttataat taaatgataa ggtagaatat
     1081 ttctgcatat aaattctggc tggcgtggaa atattcttat tggtagaaac aactacaccc
     1141 tggtcatcat cctgcctttc tctttatggt tacaatgata tacactgttt gagatgagga
     1201 taaaatactc tgagtccaaa ccgggcccct ctgctaacca tgttcatgcc ttcttctctt
     1261 tcctacagct cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaaagaat
     1321 tcctcgacgg atctatggac tacaaggatg acgatgacaa gctcgatgga ggatacccat
     1381 acgatgttcc agattacgct gatatcggat ccgaaatcgg caaaggctac actctcgagg
     1441 aggtgcttag atattctaac aaagatgtcg tggattttca tttgtcttgt gctttttgct
     1501 ctactactat ggatcataac gagaaggcca gattcataca ggctaaattg aaatgtgttg
     1561 ttagagattt tgcttttaaa ggtgcttgta ttgtgtgccg cagacagctt gcttgcaagg
     1621 aaaagctttt gcatactaga gttacagggg aggctgattt ggtagagtgc atggctggca
     1681 agaatattgt gtttgttact gtaagatgtg ttacgtgcct ggcactcctt actgcctctg
     1741 aaaagcttga tgccaaagcg tgcggcttgc catttcactt ggtgcgccac atgtggagag
     1801 gctactgcgg gttctgcaaa ccattactat aaggatccgt cgaggaattc actcctcagg
     1861 tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa tgccctggct cacaaatacc
     1921 actgagatct ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc
     1981 tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt
     2041 gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt
     2101 tggtttagag tttggcaaca tatgcccata tgctggctgc catgaacaaa ggttggctat
     2161 aaagaggtca tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag
     2221 ccttgacttg aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat
     2281 ccctaaaatt ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc
     2341 cagtcatagc tgtccctctt ctcttatgga gatccctcga cggatcggcc gcaattcgta
     2401 atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     2461 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     2521 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     2581 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     2641 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2701 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2761 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2821 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2881 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2941 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3001 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3061 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3121 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3181 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3241 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3301 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3361 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3421 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3481 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3541 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3601 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3661 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3721 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3781 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3841 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     3901 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3961 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     4021 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     4081 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4141 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     4201 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     4261 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     4321 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4381 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     4441 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4501 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     4561 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     4621 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     4681 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     4741 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     4801 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     4861 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     4921 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     4981 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct tcacgctgcc
     5041 gcaagcactc agggcgcaag ggctgctaaa ggaagcggaa cacgtagaaa gccagtccgc
     5101 agaaacggtg ctgaccccgg atgaatgtca gctactgggc tatctggaca agggaaaacg
     5161 caagcgcaaa gagaaagcag gtagcttgca gtgggcttac atggcgatag ctagactggg
     5221 cggttttatg gacagcaagc gaaccggaat tgccagctgg ggcgccctct ggtaaggttg
     5281 ggaagccctg caaagtaaac tggatggctt tcttgccgcc aaggatctga tggcgcaggg
     5341 gatcaagatc ctgcttcatc cccgtggccc gttgctcgcg tttgctggcg gtgtccccgg
     5401 aagaaatata tttgcatgtc tttagttcta tgatgacaca aaccccgccc agcgtcttgt
     5461 cattggcgaa ttcgaacacg cagatgcagt cggggcggcg cggtcccagg tccacttcgc
     5521 atattaaggt gacgcgtgtg gcctcgaaca ccgagcgacc ctgcagcgac ccgcttaaca
     5581 gcgtcaacag cgtgccgcag atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     5641 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     5701 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     5761 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     5821 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     5881 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     5941 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     6001 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     6061 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     6121 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     6181 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     6241 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     6301 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     6361 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     6421 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     6481 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     6541 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     6601 ccgggagatg ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     6661 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     6721 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     6781 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     6841 ggctcgcagc caacgtcggg gcggcaagcc ctgccatagc cacgggcccc gtgggttagg
     6901 gacggcggat cgcggccctc tagagagctt ggcccattgc atacgttgta tccatatcat
     6961 aatatgtaca tttatattgg ctcatgtcca acattaccgc catgtt
//
