LOCUS       sequence-166408-        7033 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N- Flag/HA tagged HPV 197 E6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7033)
  AUTHORS   Grace M, Munger K
  TITLE     Proteomic analysis of the gamma human papillomavirus type 197 E6 and
            E7 associated cellular proteins.
  JOURNAL   Virology. 2016 Oct 20;500:71-81. doi: 10.1016/j.virol.2016.10.010.
  PUBMED    27771561
REFERENCE   2  (bases 1 to 7033)
  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: "Virology.";
            date: "2016-10-20"; volume: "500:71-81. doi"; pages: " 
            10.1016/j.virol.2016.10.010"
COMMENT     Sequence Label: CMV-N 197 E6
FEATURES             Location/Qualifiers
     source          1..7033
                     /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(1952..1971)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2017..2072
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2071..2090)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2430..2446)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2430..2446)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2443..2465)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2454..2470
                     /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(2478..2484,2485..2502,2503..2508))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2523..2544
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2661..2678)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2832..3420)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2912..2931)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3591..4382,4383..4451))
                     /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     4214..4233
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4452..4557)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4625..4643
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4681..4703)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4803..4822
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5017..5039
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5031..5048
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5032..5048
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5459..5604
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             5661..6455
                     /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(5715..5734)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6325..6344
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6649..6668)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6693..6740
                     /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 aggcactcag 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 ccatggagcc cctgctgaag cctctgagta
     1441 ttgatgagta ttgtaactat ttccacattg attccctgta tctgaccctg ccctgtattt
     1501 tctgccacaa catcattgac agcctgcagc tgaccatctt tcagcggaga cagctgtccc
     1561 tggtgtggag gggggataat tgtttcgccg cttgcctgtc ttgtatcgca aacgtggcca
     1621 agctggagcg caacaagtac tttcagtgcc tggtcaaggg cgactatatt gaacattgta
     1681 cccagacacc cctgcagaac ctgctggtcc gatgcctgtt ctgtatggct ctgctgggcg
     1741 gggacgagaa aatcgatgtg attgccagcg gagaaaactt ctacctggtg cggggcgctt
     1801 ggaagagtgt ctgtagaaac tgtagccaga acgcttgggg agaaagtaat aactaaggat
     1861 ccgtcgagga attcactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg
     1921 ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg
     1981 acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt attttcattg
     2041 caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc
     2101 atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg
     2161 ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg
     2221 tccattcctt attccataga aaagccttga cttgaggtta gatttttttt atattttgtt
     2281 ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat
     2341 ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc
     2401 tcgacggatc ggccgcaatt cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat
     2461 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     2521 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     2581 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     2641 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     2701 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     2761 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2821 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2881 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2941 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3001 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3061 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3121 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3181 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3241 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3301 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3361 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3421 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3481 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3541 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3601 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3661 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3721 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3781 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3841 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3901 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     3961 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4021 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4081 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4141 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4201 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4261 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4321 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4381 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4441 tgaatactca tactcttcct ttttctaata ttattgaagc atttatcagg gttattgtct
     4501 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4561 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     4621 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     4681 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     4741 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     4801 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     4861 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     4921 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcggaaa gggggatgtg
     4981 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     5041 cggccagtgc caagcttcac gctgccgcaa gcactcaggg cgcaagggct gctaaaggaa
     5101 gcggaacacg tagaaagcca gtccgcagaa acggtgctga ccccggatga atgtcagcta
     5161 ctgggctatc tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg
     5221 gcttacatgg cgatagctag actgggcggt tttatggaca gcaagcgaac cggaattgcc
     5281 agctggggcg ccctctggta aggttgggaa gccctgcaaa gtaaactgga tggctttctt
     5341 gccgccaagg atctgatggc gcaggggatc aagatcctgc ttcatccccg tggcccgttg
     5401 ctcgcgtttg ctggcggtgt ccccggaaga aatatatttg catgtcttta gttctatgat
     5461 gacacaaacc ccgcccagcg tcttgtcatt ggcgaattcg aacacgcaga tgcagtcggg
     5521 gcggcgcggt cccaggtcca cttcgcatat taaggtgacg cgtgtggcct cgaacaccga
     5581 gcgaccctgc agcgacccgc ttaacagcgt caacagcgtg ccgcagatct gatcaagaga
     5641 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     5701 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     5761 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     5821 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     5881 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5941 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     6001 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     6061 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     6121 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     6181 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     6241 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     6301 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     6361 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     6421 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     6481 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     6541 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     6601 tctcatgctg gagttcttcg cccaccccgg gagatggggg aggctaactg aaacacggaa
     6661 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     6721 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     6781 tacccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     6841 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaagccctg
     6901 ccatagccac gggccccgtg ggttagggac ggcggatcgc ggccctctag agagcttggc
     6961 ccattgcata cgttgtatcc atatcataat atgtacattt atattggctc atgtccaaca
     7021 ttaccgccat gtt
//
