LOCUS       sequence-320723-        6763 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HPV18 E7 R84S.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6763)
  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 6763)
  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: p8193 MSCV-P C-FlagHA 18E7 R84S
FEATURES             Location/Qualifiers
     source          1..6763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    49..73
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(389..413)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             445..468
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             484..510
                     /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             484..510
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        523..1022
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(558..581)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     955..974
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             1043..1642
                     /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(1043..1062)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1539..1559
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             1742..2256
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    2418..2434
                     /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(2442..2448,2449..2466,2467..2472))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2487..2508
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2625..2642)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2796..3384)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2876..2895)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3555..4346,4347..4415))
                     /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     4178..4197
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4416..4520)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4588..4606
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4644..4666)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4766..4785
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4979..5001
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5129..5148
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             5348..5864
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    5928..6269
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             6336..6752
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6682..6704
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6720..6736
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 taatacgact cactataggg agacccaagc tggctagtta agctatcaac aagtttgtac
       61 aaaaaagttg gcaccatgca tggacctaag gcaacattgc aagacattgt attgcattta
      121 gagccccaaa atgaaattcc ggttgacctt ctatgtcacg agcaattaag cgactcagag
      181 gaagaaaacg atgaaataga tggagttaat catcaacatt taccagcccg acgagccgaa
      241 ccacaacgtc acacaatgtt gtgtatgtgt tgtaagtgtg aagccagaat tgagctagta
      301 gtagaaagct cagcagacga ccttagcgca ttccagcagc tgtttctgaa caccctgtcc
      361 tttgtgtgtc cgtggtgtgc atcccagcag ccaactttct tgtacaaagt ggttgatcta
      421 gagggcccgc ggttcgaact cgaggactac aaggacgacg atgacaagtc ggccgctgga
      481 ggatacccct acgacgtgcc cgactacgcc tagaattcta ccgggtaggg gaggcgcttt
      541 tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa
      601 gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct
      661 ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc
      721 cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc
      781 agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc
      841 agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc
      901 tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct
      961 gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt
     1021 cgacctgcag cccaagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg
     1081 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     1141 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     1201 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     1261 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     1321 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     1381 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     1441 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     1501 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     1561 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     1621 gacccgcaag cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag
     1681 cgcacgaccc catgcatcga taaaataaaa gattttattt agtctccaga aaaagggggg
     1741 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
     1801 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
     1861 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     1921 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
     1981 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
     2041 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
     2101 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtgtatccaa
     2161 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
     2221 tgagtgattg actacccgtc agcgggggtc tttcatgggt aacagtttct tgaagttgga
     2281 gaacaacatt ctgagggtag gagtcgaata ttaagtaatc ctgactcaat tagccactgt
     2341 tttgaatcca catactccaa tactcctgaa atagttcatt atggacagcg cagaaagagc
     2401 tggggagaat tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     2461 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     2521 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     2581 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     2641 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     2701 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     2761 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2821 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2881 taccaggcgt ttccccttgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2941 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3001 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3061 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3121 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3181 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3241 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3301 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3361 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3421 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3481 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3541 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     3601 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     3661 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     3721 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     3781 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     3841 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     3901 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     3961 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4021 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4081 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4141 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     4201 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4261 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4321 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4381 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     4441 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4501 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     4561 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     4621 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     4681 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     4741 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     4801 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg
     4861 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     4921 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     4981 cagtcacgac gttgtaaaac gacggcgcaa ggaatggtgc atgcaaggag atggcgccca
     5041 acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc
     5101 cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg
     5161 cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggcga ttagtccaat
     5221 ttgttaaaga caggatatca gtggtccagg ctctagtttt gactcaacaa tatcaccagc
     5281 tgaagcctat agagtacgag ccatagataa aataaaagat tttatttagt ctccagaaaa
     5341 aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt
     5401 gcaaggcatg gaaaatacat aactgagaat agagaagttc agatcaaggt taggaacaga
     5461 gagacagcag aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
     5521 gggccaagaa cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat
     5581 cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc
     5641 aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc
     5701 ccacaacccc tcactcggcg cgccagtcct ccgatagact gcgtcgcccg ggtacccgta
     5761 ttcccaataa agcctcttgc tgtttgcatc cgaatcgtgg actcgctgat ccttgggagg
     5821 gtctcctcag attgattgac tgcccacctc gggggtcttt catttggagg ttccaccgag
     5881 atttggagac ccctgcccag ggaccaccga cccccccgcc gggaggtaag ctggccagcg
     5941 gtcgtttcgt gtctgtctct gtctttgtgc gtgtttgtgc cggcatctaa tgtttgcgcc
     6001 tgcgtctgta ctagttagct aactagctct gtatctggcg gacccgtggt ggaactgacg
     6061 agttctgaac acccggccgc aaccctggga gacgtcccag ggactttggg ggccgttttt
     6121 gtggcccgac ctgaggaagg gagtcgatgt ggaatccgac cccgtcagga tatgtggttc
     6181 tggtaggaga cgagaaccta aaacagttcc cgcctccgtc tgaatttttg ctttcggttt
     6241 ggaaccgaag ccgcgcgtct tgtctgctgc agcgctgcag catcgttctg tgttgtctct
     6301 gtctgactgt gtttctgtat ttgtctgaaa attagggcca gactgttacc actcccttaa
     6361 gtttgacctt aggtcactgg aaagatgtcg agcggatcgc tcacaaccag tcggtagatg
     6421 tcaagaagag acgttgggtt accttctgct ctgcagaatg gccaaccttt aacgtcggat
     6481 ggccgcgaga cggcaccttt aaccgagacc tcatcaccca ggttaagatc aaggtctttt
     6541 cacctggccc gcatggacac ccagaccagg tcccctacat cgtgacctgg gaagccttgg
     6601 cttttgaccc ccctccctgg gtcaagccct ttgtacaccc taagcctccg cctcctcttc
     6661 ctccatccgc cccgtctctc ccccttgaac ctcctcgttc gaccccgcct cgatcctccc
     6721 tttatccagc cctcactcct tctctaggcg ccggaattag atc
//
