LOCUS       sequence-173641-        7051 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Flag/HA tagged HPV8 E6 K64A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7051)
  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 7051)
  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 HPV8 E6 K64A
FEATURES             Location/Qualifiers
     source          1..7051
                     /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(636..655)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    701..756
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(755..774)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1114..1130)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1114..1130)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1127..1149)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1138..1154
                     /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(1162..1168,1169..1186,1187..1192))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1207..1228
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1345..1362)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1516..2104)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1596..1615)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2275..3066,3067..3135))
                     /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     2898..2917
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3136..3240)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3308..3326
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3364..3386)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3486..3505
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3699..3721
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3713..3730
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3714..3730
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4141..4286
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             4343..5137
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDADVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4397..4416)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5007..5026
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5331..5350)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5375..5422
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     enhancer        5715..6094
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6095..6298
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6249..6269
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6295..6319
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          6410..6982
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
ORIGIN
        1 gactacaagg atgacgatga caagctcgat ggaggatacc catacgatgt tccagattac
       61 gctgatatcg gatccgacgg gcaggacaag gcttcatatt tagacactaa taaggacgag
      121 ctaccctcta ctattaaaga gttagctgcg gctttaggta ttccattgca ggactgttta
      181 gtaccgtgca acttttgtgg taactttttg gatttcttag aactgtgtga gtttgacaaa
      241 aagagactgt gcctaatttg ggcaaattac gttgttactg cgtgttgtcg ttgttgttgt
      301 gtagcaaccg caacgtttga atttaatgaa tattatgagc aaactgtgct aggcagagat
      361 attgaattag ctacaggacg ttcaattttt gagatagacg ttaggtgtca aaactgcttg
      421 tcatttttgg atatcataga gaaattagat tgctgtggga gaggccgtcc ctttcataaa
      481 gttagaggag gctggaaagg agtttgcagg ctttgtaagc atttgtatca tgattggtaa
      541 agatccgtcg aggaattcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt
      601 gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat
      661 ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc
      721 attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca
      781 aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg
      841 ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct
      901 gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt
      961 tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc
     1021 agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga
     1081 tccctcgacg gatcggccgc aattcgtaat catgtcatag ctgtttcctg tgtgaaattg
     1141 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     1201 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     1261 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     1321 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     1381 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     1441 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1501 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1561 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1621 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1681 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1741 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1801 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1861 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1921 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     1981 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2041 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2101 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2161 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2221 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     2281 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     2341 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     2401 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     2461 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     2521 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     2581 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     2641 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     2701 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     2761 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     2821 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     2881 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     2941 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3001 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3061 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     3121 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     3181 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     3241 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     3301 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     3361 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     3421 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     3481 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     3541 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     3601 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     3661 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     3721 gacggccagt gccaagcttc acgctgccgc aagcactcag ggcgcaaggg ctgctaaagg
     3781 aagcggaaca cgtagaaagc cagtccgcag aaacggtgct gaccccggat gaatgtcagc
     3841 tactgggcta tctggacaag ggaaaacgca agcgcaaaga gaaagcaggt agcttgcagt
     3901 gggcttacat ggcgatagct agactgggcg gttttatgga cagcaagcga accggaattg
     3961 ccagctgggg cgccctctgg taaggttggg aagccctgca aagtaaactg gatggctttc
     4021 ttgccgccaa ggatctgatg gcgcagggga tcaagatcct gcttcatccc cgtggcccgt
     4081 tgctcgcgtt tgctggcggt gtccccggaa gaaatatatt tgcatgtctt tagttctatg
     4141 atgacacaaa ccccgcccag cgtcttgtca ttggcgaatt cgaacacgca gatgcagtcg
     4201 gggcggcgcg gtcccaggtc cacttcgcat attaaggtga cgcgtgtggc ctcgaacacc
     4261 gagcgaccct gcagcgaccc gcttaacagc gtcaacagcg tgccgcagat ctgatcaaga
     4321 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4381 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4441 tgccgacgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4501 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     4561 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4621 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4681 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4741 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4801 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4861 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4921 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4981 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     5041 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5101 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     5161 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5221 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5281 gatctcatgc tggagttctt cgcccacccc gggagatggg ggaggctaac tgaaacacgg
     5341 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     5401 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     5461 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     5521 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaagccct
     5581 gccatagcca cgggccccgt gggttaggga cggcggatcg cggccctcta gagagcttgg
     5641 cccattgcat acgttgtatc catatcataa tatgtacatt tatattggct catgtccaac
     5701 attaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
     5761 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     5821 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     5881 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     5941 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     6001 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     6061 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     6121 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     6181 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     6241 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg
     6301 tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag
     6361 acaccgggac cgatccagcc tccggtcgac cgatcctgag aacttcaggg tgagtttggg
     6421 gacccttgat tgttctttct ttttcgctat tgtaaaattc atgttatatg gagggggcaa
     6481 agttttcagg gtgttgttta gaatgggaag atgtcccttg tatcaccatg gaccctcatg
     6541 ataattttgt ttctttcact ttctactctg ttgacaacca ttgtctcctc ttattttctt
     6601 ttcattttct gtaacttttt cgttaaactt tagcttgcat ttgtaacgaa tttttaaatt
     6661 cacttttgtt tatttgtcag attgtaagta ctttctctaa tcactttttt ttcaaggcaa
     6721 tcagggtata ttatattgta cttcagcaca gttttagaga acaattgtta taattaaatg
     6781 ataaggtaga atatttctgc atataaattc tggctggcgt ggaaatattc ttattggtag
     6841 aaacaactac accctggtca tcatcctgcc tttctcttta tggttacaat gatatacact
     6901 gtttgagatg aggataaaat actctgagtc caaaccgggc ccctctgcta accatgttca
     6961 tgccttcttc tctttcctac agctcctggg caacgtgctg gttgttgtgc tgtctcatca
     7021 ttttggcaaa gaattcctcg acggatctat g
//
