LOCUS       sequence-166375-        6890 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-Flag/HA tagged HPV 16 E7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6890)
  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 6890)
  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 16 E7
FEATURES             Location/Qualifiers
     source          1..6890
                     /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             1649..1672
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1685..1711
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1810..1829)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1875..1930
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1929..1948)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2287..2303)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2287..2303)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2300..2322)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2311..2327
                     /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(2335..2341,2342..2359,2360..2365))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2380..2401
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2518..2535)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2689..3277)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2769..2788)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3448..4239,4240..4308))
                     /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     4071..4090
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4309..4414)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4482..4500
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4538..4560)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4660..4679
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4874..4896
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4888..4905
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4889..4905
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5316..5461
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             5518..6312
                     /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(5572..5591)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6182..6201
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6506..6525)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6550..6597
                     /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 atccgattcg agaccatgca tggagataca cctacattgc atgaatatat
     1381 gttagatttg caaccagaga caactgatct ctactgttat gagcaattaa atgacagctc
     1441 agaggaggag gatgaaatag atggtccagc tggacaagca gaaccggaca gagcccatta
     1501 caatattgta accttttgtt gcaagtgtga ctctacgctt cggttgtgcg tacaaagcac
     1561 acacgtagac attcgtactt tggaagacct gttaatgggc acactaggaa ttgtgtgccc
     1621 catctgttct cagaaaccag cggccatcga ctacaaggat gacgatgaca agctcgatgg
     1681 aggataccca tacgatgttc cagattacgc ttagagatcc gtcgaggaat tcactcctca
     1741 ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg ctcacaaata
     1801 ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca
     1861 tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt
     1921 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta
     1981 tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct
     2041 ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa
     2101 agccttgact tgaggtagat tttttttata ttttgttttg tgttattttt ttctttaaca
     2161 tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc
     2221 ccagtcatag ctgtccctct tctcttatgg agatccctcg acggatcggc cgcaattcgt
     2281 aatcatgtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     2341 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     2401 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     2461 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     2521 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     2581 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     2641 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2701 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2761 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2821 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2881 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2941 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3001 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3061 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3121 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3181 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3241 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3301 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3361 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3421 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3481 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3541 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     3601 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     3661 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3721 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     3781 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3841 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3901 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     3961 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4021 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4081 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4141 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4201 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4261 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4321 tctaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     4381 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     4441 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     4501 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     4561 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     4621 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     4681 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     4741 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     4801 cttcgctatt acgccagctg gcggaaaggg ggatgtgctg caaggcgatt aagttgggta
     4861 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gcttcacgct
     4921 gccgcaagca ctcagggcgc aagggctgct aaaggaagcg gaacacgtag aaagccagtc
     4981 cgcagaaacg gtgctgaccc cggatgaatg tcagctactg ggctatctgg acaagggaaa
     5041 acgcaagcgc aaagagaaag caggtagctt gcagtgggct tacatggcga tagctagact
     5101 gggcggtttt atggacagca agcgaaccgg aattgccagc tggggcgccc tctggtaagg
     5161 ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc gccaaggatc tgatggcgca
     5221 ggggatcaag atcctgcttc atccccgtgg cccgttgctc gcgtttgctg gcggtgtccc
     5281 cggaagaaat atatttgcat gtctttagtt ctatgatgac acaaaccccg cccagcgtct
     5341 tgtcattggc gaattcgaac acgcagatgc agtcggggcg gcgcggtccc aggtccactt
     5401 cgcatattaa ggtgacgcgt gtggcctcga acaccgagcg accctgcagc gacccgctta
     5461 acagcgtcaa cagcgtgccg cagatctgat caagagacag gatgaggatc gtttcgcatg
     5521 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     5581 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     5641 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     5701 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     5761 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     5821 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     5881 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     5941 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     6001 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     6061 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     6121 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     6181 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     6241 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     6301 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     6361 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     6421 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     6481 accccgggag atgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     6541 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     6601 aacgcggggt tcggtcccag ggctggcact ctgtcgatac ccccaccgag accccattgg
     6661 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     6721 ccagggctcg cagccaacgt cggggcggca agccctgcca tagccacggg ccccgtgggt
     6781 tagggacggc ggatcgcggc cctctagaga gcttggccca ttgcatacgt tgtatccata
     6841 tcataatatg tacatttata ttggctcatg tccaacatta ccgccatgtt
//
