LOCUS       sequence-173646-        7036 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Flag/HA tagged HPV8 E6 del132-136.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7036)
  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 7036)
  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 del132-136
FEATURES             Location/Qualifiers
     source          1..7036
                     /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(621..640)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    686..741
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(740..759)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1099..1115)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1099..1115)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1112..1134)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1123..1139
                     /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(1147..1153,1154..1171,1172..1177))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1192..1213
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1330..1347)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1501..2089)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1581..1600)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2260..3051,3052..3120))
                     /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     2883..2902
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3121..3225)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3293..3311
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3349..3371)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3471..3490
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3684..3706
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3698..3715
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3699..3715
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4126..4271
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             4328..5122
                     /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(4382..4401)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4992..5011
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5316..5335)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5360..5407
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     enhancer        5700..6079
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6080..6283
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6234..6254
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6280..6304
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          6395..6967
                     /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 gaaaaattac gttgttactg cgtgttgtcg ttgttgttgt
      301 gtagcaaccg caacgtttga atttaatgaa tattatgagc aaactgtgct aggcagagat
      361 attgaattag ctacaggacg ttcaattttt gagatagacg ttaggtgtca aaactgcttg
      421 tcatttttgg atatcataga gaaattagat tgctgtggga gaggcgttag aggaggctgg
      481 aaaggagttt gcaggctttg taagcatttg tatcatgatt ggtaaagatc cgtcgaggaa
      541 ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg
      601 gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag
      661 ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt
      721 tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat
      781 cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac
      841 aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta
      901 ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt
      961 ttttctttaa catccctaaa attttcctta catgttttac tagccagatt tttcctcctc
     1021 tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct cgacggatcg
     1081 gccgcaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     1141 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     1201 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     1261 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     1321 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     1381 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     1441 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     1501 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     1561 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     1621 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     1681 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     1741 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     1801 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     1861 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     1921 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     1981 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2041 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2101 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2161 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2221 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2281 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     2341 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     2401 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     2461 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     2521 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     2581 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     2641 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     2701 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     2761 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     2821 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     2881 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     2941 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     3001 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     3061 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     3121 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     3181 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     3241 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     3301 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     3361 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     3421 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     3481 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     3541 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     3601 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     3661 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa
     3721 gcttcacgct gccgcaagca ctcagggcgc aagggctgct aaaggaagcg gaacacgtag
     3781 aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg tcagctactg ggctatctgg
     3841 acaagggaaa acgcaagcgc aaagagaaag caggtagctt gcagtgggct tacatggcga
     3901 tagctagact gggcggtttt atggacagca agcgaaccgg aattgccagc tggggcgccc
     3961 tctggtaagg ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc gccaaggatc
     4021 tgatggcgca ggggatcaag atcctgcttc atccccgtgg cccgttgctc gcgtttgctg
     4081 gcggtgtccc cggaagaaat atatttgcat gtctttagtt ctatgatgac acaaaccccg
     4141 cccagcgtct tgtcattggc gaattcgaac acgcagatgc agtcggggcg gcgcggtccc
     4201 aggtccactt cgcatattaa ggtgacgcgt gtggcctcga acaccgagcg accctgcagc
     4261 gacccgctta acagcgtcaa cagcgtgccg cagatctgat caagagacag gatgaggatc
     4321 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4381 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4441 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4501 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4561 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4621 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4681 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4741 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4801 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     4861 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4921 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4981 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     5041 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     5101 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     5161 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     5221 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     5281 ttcttcgccc accccgggag atgggggagg ctaactgaaa cacggaagga gacaataccg
     5341 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     5401 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     5461 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     5521 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcaa gccctgccat agccacgggc
     5581 cccgtgggtt agggacggcg gatcgcggcc ctctagagag cttggcccat tgcatacgtt
     5641 gtatccatat cataatatgt acatttatat tggctcatgt ccaacattac cgccatgttg
     5701 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     5761 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     5821 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     5881 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     5941 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     6001 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     6061 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     6121 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     6181 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     6241 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca
     6301 gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc
     6361 cagcctccgg tcgaccgatc ctgagaactt cagggtgagt ttggggaccc ttgattgttc
     6421 tttctttttc gctattgtaa aattcatgtt atatggaggg ggcaaagttt tcagggtgtt
     6481 gtttagaatg ggaagatgtc ccttgtatca ccatggaccc tcatgataat tttgtttctt
     6541 tcactttcta ctctgttgac aaccattgtc tcctcttatt ttcttttcat tttctgtaac
     6601 tttttcgtta aactttagct tgcatttgta acgaattttt aaattcactt ttgtttattt
     6661 gtcagattgt aagtactttc tctaatcact tttttttcaa ggcaatcagg gtatattata
     6721 ttgtacttca gcacagtttt agagaacaat tgttataatt aaatgataag gtagaatatt
     6781 tctgcatata aattctggct ggcgtggaaa tattcttatt ggtagaaaca actacaccct
     6841 ggtcatcatc ctgcctttct ctttatggtt acaatgatat acactgtttg agatgaggat
     6901 aaaatactct gagtccaaac cgggcccctc tgctaaccat gttcatgcct tcttctcttt
     6961 cctacagctc ctgggcaacg tgctggttgt tgtgctgtct catcattttg gcaaagaatt
     7021 cctcgacgga tctatg
//
