LOCUS       sequence-166411-        6874 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-Flag/HA tagged HPV 197 E7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6874)
  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 6874)
  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 E7
FEATURES             Location/Qualifiers
     source          1..6874
                     /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(1793..1812)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1858..1913
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1912..1931)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2271..2287)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2271..2287)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2284..2306)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2295..2311
                     /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(2319..2325,2326..2343,2344..2349))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2364..2385
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2502..2519)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2673..3261)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2753..2772)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3432..4223,4224..4292))
                     /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     4055..4074
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4293..4398)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4466..4484
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4522..4544)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4644..4663
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4858..4880
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4872..4889
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4873..4889
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5300..5445
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             5502..6296
                     /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(5556..5575)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6166..6185
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6490..6509)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6534..6581
                     /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 ccatgcacgg ggaaagagca actattagag
     1441 atgtggtcct ggaactgaca aaggtcgcct gtaactcaga agtcctgttt gaagagagcc
     1501 tgagctccga ctccgaggaa gagcacgaaa tcttcaagat tgagacccga tgcgataact
     1561 gtcgaagagg catcagggtg tgcgtccgcg catctaatca ggggattagt agactgcagt
     1621 ggctgctgga acatgacatt aggtttctgt gccccgcttg tagccgcaat ctgtttcagc
     1681 atggaaggtt tcagtaagga tccgtcgagg aattcactcc tcaggtgcag gctgcctatc
     1741 agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc
     1801 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctggctaat
     1861 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     1921 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     1981 caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt
     2041 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt
     2101 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct
     2161 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc
     2221 ctcttctctt atggagatcc ctcgacggat cggccgcaat tcgtaatcat gtcatagctg
     2281 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     2341 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     2401 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     2461 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     2521 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2581 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2641 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2701 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2761 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2821 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2881 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2941 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3001 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3061 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3121 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3181 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3241 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3301 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3361 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3421 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3481 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3541 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     3601 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3661 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3721 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     3781 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     3841 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     3901 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     3961 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4021 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4081 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4141 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4201 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4261 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttctaat attattgaag
     4321 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4381 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     4441 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     4501 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     4561 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     4621 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     4681 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     4741 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     4801 gctggcggaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     4861 agtcacgacg ttgtaaaacg acggccagtg ccaagcttca cgctgccgca agcactcagg
     4921 gcgcaagggc tgctaaagga agcggaacac gtagaaagcc agtccgcaga aacggtgctg
     4981 accccggatg aatgtcagct actgggctat ctggacaagg gaaaacgcaa gcgcaaagag
     5041 aaagcaggta gcttgcagtg ggcttacatg gcgatagcta gactgggcgg ttttatggac
     5101 agcaagcgaa ccggaattgc cagctggggc gccctctggt aaggttggga agccctgcaa
     5161 agtaaactgg atggctttct tgccgccaag gatctgatgg cgcaggggat caagatcctg
     5221 cttcatcccc gtggcccgtt gctcgcgttt gctggcggtg tccccggaag aaatatattt
     5281 gcatgtcttt agttctatga tgacacaaac cccgcccagc gtcttgtcat tggcgaattc
     5341 gaacacgcag atgcagtcgg ggcggcgcgg tcccaggtcc acttcgcata ttaaggtgac
     5401 gcgtgtggcc tcgaacaccg agcgaccctg cagcgacccg cttaacagcg tcaacagcgt
     5461 gccgcagatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     5521 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     5581 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     5641 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     5701 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     5761 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     5821 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     5881 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     5941 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     6001 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     6061 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     6121 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     6181 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     6241 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     6301 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     6361 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     6421 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccaccccg ggagatgggg
     6481 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     6541 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     6601 ccagggctgg cactctgtcg atacccccac cgagacccca ttggggccaa tacgcccgcg
     6661 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     6721 acgtcggggc ggcaagccct gccatagcca cgggccccgt gggttaggga cggcggatcg
     6781 cggccctcta gagagcttgg cccattgcat acgttgtatc catatcataa tatgtacatt
     6841 tatattggct catgtccaac attaccgcca tgtt
//
