LOCUS       sequence-180206-        7051 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Flag/HA tagged HPV8 E6 R138S.
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 Apr 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 R138S
FEATURES             Location/Qualifiers
     source          1..7051
                     /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(1973..1992)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2038..2093
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2092..2111)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2451..2467)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2451..2467)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2464..2486)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2475..2491
                     /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(2499..2505,2506..2523,2524..2529))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2544..2565
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2682..2699)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2853..3441)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2933..2952)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3612..4403,4404..4472))
                     /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     4235..4254
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4473..4577)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4645..4663
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4701..4723)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4823..4842
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5036..5058
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5050..5067
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5051..5067
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5478..5623
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             5680..6474
                     /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(5734..5753)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6344..6363
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6668..6687)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6712..6759
                     /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 ccgacgggca ggacaaggct tcatatttag
     1441 acactaataa ggacgagcta ccctctacta ttaaagagtt agctgcggct ttaggtattc
     1501 cattgcagga ctgtttagta ccgtgcaact tttgtggtaa ctttttggat ttcttagaac
     1561 tgtgtgagtt tgacaaaaag agactgtgcc taatttggaa aaattacgtt gttactgcgt
     1621 gttgtcgttg ttgttgtgta gcaaccgcaa cgtttgaatt taatgaatat tatgagcaaa
     1681 ctgtgctagg cagagatatt gaattagcta caggacgttc aatttttgag atagacgtta
     1741 ggtgtcaaaa ctgcttgtca tttttggata tcatagagaa attagattgc tgtgggagag
     1801 gccgtccctt tcataaagtt agcggaggct ggaaaggagt ttgcaggctt tgtaagcatt
     1861 tgtatcatga ttggtaaaga tccgtcgagg aattcactcc tcaggtgcag gctgcctatc
     1921 agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc
     1981 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctggctaat
     2041 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     2101 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     2161 caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt
     2221 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt
     2281 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct
     2341 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc
     2401 ctcttctctt atggagatcc ctcgacggat cggccgcaat tcgtaatcat gtcatagctg
     2461 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     2521 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     2581 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     2641 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     2701 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2761 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2821 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2881 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2941 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3001 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3061 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3121 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3181 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3241 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3301 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3361 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3421 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3481 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3541 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3601 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3661 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3721 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     3781 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3841 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3901 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     3961 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4021 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4081 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4141 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4201 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4261 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4321 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4381 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4441 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4501 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4561 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     4621 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     4681 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     4741 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     4801 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     4861 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     4921 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     4981 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     5041 tcacgacgtt gtaaaacgac ggccagtgcc aagcttcacg ctgccgcaag cactcagggc
     5101 gcaagggctg ctaaaggaag cggaacacgt agaaagccag tccgcagaaa cggtgctgac
     5161 cccggatgaa tgtcagctac tgggctatct ggacaaggga aaacgcaagc gcaaagagaa
     5221 agcaggtagc ttgcagtggg cttacatggc gatagctaga ctgggcggtt ttatggacag
     5281 caagcgaacc ggaattgcca gctggggcgc cctctggtaa ggttgggaag ccctgcaaag
     5341 taaactggat ggctttcttg ccgccaagga tctgatggcg caggggatca agatcctgct
     5401 tcatccccgt ggcccgttgc tcgcgtttgc tggcggtgtc cccggaagaa atatatttgc
     5461 atgtctttag ttctatgatg acacaaaccc cgcccagcgt cttgtcattg gcgaattcga
     5521 acacgcagat gcagtcgggg cggcgcggtc ccaggtccac ttcgcatatt aaggtgacgc
     5581 gtgtggcctc gaacaccgag cgaccctgca gcgacccgct taacagcgtc aacagcgtgc
     5641 cgcagatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     5701 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     5761 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     5821 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     5881 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     5941 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     6001 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     6061 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     6121 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     6181 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     6241 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     6301 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     6361 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     6421 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     6481 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     6541 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     6601 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccggg agatggggga
     6661 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     6721 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     6781 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     6841 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     6901 tcggggcggc aagccctgcc atagccacgg gccccgtggg ttagggacgg cggatcgcgg
     6961 ccctctagag agcttggccc attgcatacg ttgtatccat atcataatat gtacatttat
     7021 attggctcat gtccaacatt accgccatgt t
//
