LOCUS       sequence-323448-        6763 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HPV18 E7 D10K.
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: p8141 MSCV-P C-FlagHA 18E7 D10K
FEATURES             Location/Qualifiers
     source          1..6763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             63..479
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     409..431
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     447..463
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     491..510
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        491..509
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    539..563
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(879..903)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             935..958
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             974..1000
                     /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             974..1000
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        1013..1512
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1048..1071)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1445..1464
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             1533..2132
                     /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(1533..1552)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2029..2049
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             2232..2746
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    2908..2924
                     /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(2932..2938,2939..2956,2957..2962))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2977..2998
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3115..3132)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3286..3874)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3366..3385)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4045..4836,4837..4905))
                     /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     4668..4687
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4906..5010)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5078..5096
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5134..5156)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5256..5275
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5469..5491
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5619..5638
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             5838..6354
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6418..6759
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
ORIGIN
        1 gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt
       61 agggccagac tgttaccact cccttaagtt tgaccttagg tcactggaaa gatgtcgagc
      121 ggatcgctca caaccagtcg gtagatgtca agaagagacg ttgggttacc ttctgctctg
      181 cagaatggcc aacctttaac gtcggatggc cgcgagacgg cacctttaac cgagacctca
      241 tcacccaggt taagatcaag gtcttttcac ctggcccgca tggacaccca gaccaggtcc
      301 cctacatcgt gacctgggaa gccttggctt ttgacccccc tccctgggtc aagccctttg
      361 tacaccctaa gcctccgcct cctcttcctc catccgcccc gtctctcccc cttgaacctc
      421 ctcgttcgac cccgcctcga tcctcccttt atccagccct cactccttct ctaggcgccg
      481 gaattagatc taatacgact cactataggg agacccaagc tggctagtta agctatcaac
      541 aagtttgtac aaaaaagttg gcaccatgca tggacctaag gcaacattgc aaaaaattgt
      601 attgcattta gagccccaaa atgaaattcc ggttgacctt ctatgtcacg agcaattaag
      661 cgactcagag gaagaaaacg atgaaataga tggagttaat catcaacatt taccagcccg
      721 acgagccgaa ccacaacgtc acacaatgtt gtgtatgtgt tgtaagtgtg aagccagaat
      781 tgagctagta gtagaaagct cagcagacga ccttcgagca ttccagcagc tgtttctgaa
      841 caccctgtcc tttgtgtgtc cgtggtgtgc atcccagcag ccaactttct tgtacaaagt
      901 ggttgatcta gagggcccgc ggttcgaact cgaggactac aaggacgacg atgacaagtc
      961 ggccgctgga ggatacccct acgacgtgcc cgactacgcc tagaattcta ccgggtaggg
     1021 gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg
     1081 cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg
     1141 gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc
     1201 ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact
     1261 agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc
     1321 ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc
     1381 gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc
     1441 ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct
     1501 ccgggccttt cgacctgcag cccaagctta ccatgaccga gtacaagccc acggtgcgcc
     1561 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact
     1621 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc
     1681 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     1741 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     1801 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     1861 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     1921 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     1981 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     2041 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     2101 cctggtgcat gacccgcaag cccggtgcct gacgcccgcc ccacgacccg cagcgcccga
     2161 ccgaaaggag cgcacgaccc catgcatcga taaaataaaa gattttattt agtctccaga
     2221 aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat
     2281 tttgcaaggc atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac
     2341 agagagacag cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc
     2401 tcagggccaa gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac
     2461 catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta
     2521 accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag
     2581 agcccacaac ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc
     2641 gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga
     2701 gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatgggt aacagtttct
     2761 tgaagttgga gaacaacatt ctgagggtag gagtcgaata ttaagtaatc ctgactcaat
     2821 tagccactgt tttgaatcca catactccaa tactcctgaa atagttcatt atggacagcg
     2881 cagaaagagc tggggagaat tgtgaaattg ttatccgctc acaattccac acaacatacg
     2941 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     3001 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     3061 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     3121 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3181 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3241 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3301 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3361 actataaaga taccaggcgt ttccccttgg aagctccctc gtgcgctctc ctgttccgac
     3421 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3481 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3541 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3601 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3661 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3721 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3781 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3841 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3901 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3961 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4021 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4081 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4141 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4201 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4261 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4321 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4381 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4441 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4501 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4561 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4621 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4681 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4741 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4801 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4861 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4921 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4981 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     5041 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     5101 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     5161 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     5221 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     5281 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     5341 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     5401 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     5461 agggttttcc cagtcacgac gttgtaaaac gacggcgcaa ggaatggtgc atgcaaggag
     5521 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
     5581 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg
     5641 ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggcga
     5701 ttagtccaat ttgttaaaga caggatatca gtggtccagg ctctagtttt gactcaacaa
     5761 tatcaccagc tgaagcctat agagtacgag ccatagataa aataaaagat tttatttagt
     5821 ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta
     5881 acgccatttt gcaaggcatg gaaaatacat aactgagaat agagaagttc agatcaaggt
     5941 taggaacaga gagacagcag aatatgggcc aaacaggata tctgtggtaa gcagttcctg
     6001 ccccggctca gggccaagaa cagatggtcc ccagatgcgg tcccgccctc agcagtttct
     6061 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
     6121 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
     6181 ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgatagact gcgtcgcccg
     6241 ggtacccgta ttcccaataa agcctcttgc tgtttgcatc cgaatcgtgg actcgctgat
     6301 ccttgggagg gtctcctcag attgattgac tgcccacctc gggggtcttt catttggagg
     6361 ttccaccgag atttggagac ccctgcccag ggaccaccga cccccccgcc gggaggtaag
     6421 ctggccagcg gtcgtttcgt gtctgtctct gtctttgtgc gtgtttgtgc cggcatctaa
     6481 tgtttgcgcc tgcgtctgta ctagttagct aactagctct gtatctggcg gacccgtggt
     6541 ggaactgacg agttctgaac acccggccgc aaccctggga gacgtcccag ggactttggg
     6601 ggccgttttt gtggcccgac ctgaggaagg gagtcgatgt ggaatccgac cccgtcagga
     6661 tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc tgaatttttg
     6721 ctttcggttt ggaaccgaag ccgcgcgtct tgtctgctgc agc
//
