LOCUS       sequence-322809-        6742 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HPV31 E7 R77S.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6742)
  AUTHORS   Hatterschide J, Brantly AC, Grace M, Munger K, White EA
  TITLE     A Conserved Amino Acid in the C Terminus of Human Papillomavirus E7 
            Mediates Binding to PTPN14 and Repression of Epithelial 
            Differentiation.
  JOURNAL   J Virol. 2020 Aug 17;94(17). pii: JVI.01024-20. doi: 
            10.1128/JVI.01024-20. Print 2020 Aug 17.
  PUBMED    32581101
REFERENCE   2  (bases 1 to 6742)
  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: "J Virol."; 
            date: "2020-08-17"; volume: "94(17). pii: JVI.01024-20. doi"; pages:
            " 10.1128/JVI.01024-20. Print 2020 Aug 17"
COMMENT     Sequence Label: p8194 MSCV-P C-FlagHA 31E7 R77S
FEATURES             Location/Qualifiers
     source          1..6742
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    64..405
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             472..888
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     818..840
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     856..872
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     900..919
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        900..918
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    948..972
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1267..1291)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1323..1346
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1362..1388
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             1362..1388
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        1401..1900
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1436..1459)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1833..1852
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             1921..2520
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1921..1940)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2417..2437
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             2620..3134
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3296..3312
                     /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(3320..3326,3327..3344,3345..3350))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3365..3386
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3503..3520)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3674..4262)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3754..3773)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4433..5224,5225..5293))
                     /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     5056..5075
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5294..5398)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5466..5484
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5522..5544)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5644..5663
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5857..5879
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6007..6026
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6226..6742
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tggaggttcc accgagattt ggagacccct gcccagggac caccgacccc cccgccggga
       61 ggtaagctgg ccagcggtcg tttcgtgtct gtctctgtct ttgtgcgtgt ttgtgccggc
      121 atctaatgtt tgcgcctgcg tctgtactag ttagctaact agctctgtat ctggcggacc
      181 cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac
      241 tttgggggcc gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg
      301 tcaggatatg tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa
      361 tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc
      421 gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact
      481 gttaccactc ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac
      541 aaccagtcgg tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca
      601 acctttaacg tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt
      661 aagatcaagg tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg
      721 acctgggaag ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag
      781 cctccgcctc ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc
      841 ccgcctcgat cctcccttta tccagccctc actccttctc taggcgccgg aattagatct
      901 aatacgactc actataggga gacccaagct ggctagttaa gctatcaaca agtttgtaca
      961 aaaaagttgg caccatgcgt ggagaaacac ctacgttgca agactatgtg ttagatttgc
     1021 aacctgaggc aactgacctc cactgttatg agcaattacc cgacagctca gatgaggagg
     1081 atgtcataga cagtccagct ggacaagcag aaccggacac atccaattac aatatcgtta
     1141 ccttttgttg tcagtgtaag tctacacttc gtttgtgtgt acagagcaca caagtagata
     1201 ttagcatatt gcaagagctg ttaatgggct catttggaat cgtgtgcccc aactgttcta
     1261 ctagactgcc aactttcttg tacaaagtgg ttgatctaga gggcccgcgg ttcgaactcg
     1321 aggactacaa ggacgacgat gacaagtcgg ccgctggagg atacccctac gacgtgcccg
     1381 actacgccta gaattctacc gggtagggga ggcgcttttc ccaaggcagt ctggagcatg
     1441 cgctttagca gccccgctgg gcacttggcg ctacacaagt ggcctctggc ctcgcacaca
     1501 ttccacatcc accggtaggc gccaaccggc tccgttcttt ggtggcccct tcgcgccacc
     1561 ttctactcct cccctagtca ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac
     1621 gtgacaaatg gaagtagcac gtctcactag tctcgtgcag atggacagca ccgctgagca
     1681 atggaagcgg gtaggccttt ggggcagcgg ccaatagcag ctttgctcct tcgctttctg
     1741 ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg
     1801 ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc
     1861 tgccgcgctg ttctcctctt cctcatctcc gggcctttcg acctgcagcc caagcttacc
     1921 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
     1981 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     2041 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     2101 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     2161 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     2221 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     2281 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     2341 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     2401 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     2461 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     2521 cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg cacgacccca tgcatcgata
     2581 aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg
     2641 tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaataca taactgagaa
     2701 tagagaagtt cagatcaagg ttaggaacag agagacagca gaatatgggc caaacaggat
     2761 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
     2821 gtcccgccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
     2881 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
     2941 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
     3001 tccgatagac tgcgtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
     3061 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
     3121 cgggggtctt tcatgggtaa cagtttcttg aagttggaga acaacattct gagggtagga
     3181 gtcgaatatt aagtaatcct gactcaatta gccactgttt tgaatccaca tactccaata
     3241 ctcctgaaat agttcattat ggacagcgca gaaagagctg gggagaattg tgaaattgtt
     3301 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     3361 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     3421 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     3481 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3541 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3601 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3661 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3721 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccttggaa
     3781 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3841 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3901 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3961 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4021 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4081 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4141 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4201 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4261 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4321 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4381 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     4441 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4501 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     4561 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4621 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     4681 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4741 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4801 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4861 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4921 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4981 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5041 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5101 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5161 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     5221 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     5281 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5341 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5401 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     5461 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     5521 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     5581 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     5641 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     5701 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
     5761 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     5821 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     5881 cggcgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc
     5941 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
     6001 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
     6061 ggccacgatg cgtccggcgt agaggcgatt agtccaattt gttaaagaca ggatatcagt
     6121 ggtccaggct ctagttttga ctcaacaata tcaccagctg aagcctatag agtacgagcc
     6181 atagataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac
     6241 ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga aaatacataa
     6301 ctgagaatag agaagttcag atcaaggtta ggaacagaga gacagcagaa tatgggccaa
     6361 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc
     6421 agatgcggtc ccgccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca
     6481 aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct
     6541 gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggcgcg
     6601 ccagtcctcc gatagactgc gtcgcccggg tacccgtatt cccaataaag cctcttgctg
     6661 tttgcatccg aatcgtggac tcgctgatcc ttgggagggt ctcctcagat tgattgactg
     6721 cccacctcgg gggtctttca tt
//
