LOCUS       sequence-221694-        7030 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-Flag/HA tagged HPV 197 E6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7030)
  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 7030)
  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-C 197 E6
FEATURES             Location/Qualifiers
     source          1..7030
                     /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             1791..1814
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1827..1853
                     /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..4556)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4624..4642
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4680..4702)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4802..4821
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5015..5037
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5029..5046
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5030..5046
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5457..5602
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             5659..6453
                     /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(5713..5732)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6323..6342
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6647..6666)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6691..6738
                     /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 atccatggag cccctgctga agcctctgag tattgatgag tattgtaact
     1381 atttccacat tgattccctg tatctgaccc tgccctgtat tttctgccac aacatcattg
     1441 acagcctgca gctgaccatc tttcagcgga gacagctgtc cctggtgtgg aggggggata
     1501 attgtttcgc cgcttgcctg tcttgtatcg caaacgtggc caagctggag cgcaacaagt
     1561 actttcagtg cctggtcaag ggcgactata ttgaacattg tacccagaca cccctgcaga
     1621 acctgctggt ccgatgcctg ttctgtatgg ctctgctggg cggggacgag aaaatcgatg
     1681 tgattgccag cggagaaaac ttctacctgg tgcggggcgc ttggaagagt gtctgtagaa
     1741 actgtagcca gaacgcttgg ggagaaagta ataactcagg atccgatatc gactacaagg
     1801 atgacgatga caagctcgat ggaggatacc catacgatgt tccagattac gcttagagat
     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 ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4501 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4561 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     4621 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     4681 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     4741 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     4801 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     4861 tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg
     4921 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     4981 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     5041 gccagtgcca agcttcacgc tgccgcaagc actcagggcg caagggctgc taaaggaagc
     5101 ggaacacgta gaaagccagt ccgcagaaac ggtgctgacc ccggatgaat gtcagctact
     5161 gggctatctg gacaagggaa aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc
     5221 ttacatggcg atagctagac tgggcggttt tatggacagc aagcgaaccg gaattgccag
     5281 ctggggcgcc ctctggtaag gttgggaagc cctgcaaagt aaactggatg gctttcttgc
     5341 cgccaaggat ctgatggcgc aggggatcaa gatcctgctt catccccgtg gcccgttgct
     5401 cgcgtttgct ggcggtgtcc ccggaagaaa tatatttgca tgtctttagt tctatgatga
     5461 cacaaacccc gcccagcgtc ttgtcattgg cgaattcgaa cacgcagatg cagtcggggc
     5521 ggcgcggtcc caggtccact tcgcatatta aggtgacgcg tgtggcctcg aacaccgagc
     5581 gaccctgcag cgacccgctt aacagcgtca acagcgtgcc gcagatctga tcaagagaca
     5641 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     5701 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     5761 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     5821 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     5881 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     5941 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     6001 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     6061 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     6121 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     6181 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     6241 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     6301 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     6361 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     6421 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     6481 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     6541 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     6601 tcatgctgga gttcttcgcc caccccggga gatgggggag gctaactgaa acacggaagg
     6661 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     6721 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     6781 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     6841 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca agccctgcca
     6901 tagccacggg ccccgtgggt tagggacggc ggatcgcggc cctctagaga gcttggccca
     6961 ttgcatacgt tgtatccata tcataatatg tacatttata ttggctcatg tccaacatta
     7021 ccgccatgtt
//
