LOCUS       sequence-323447-        6730 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HPV16 E7 ∆DLYC.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6730)
  AUTHORS   Hatterschide J, Bohidar AE, Grace M, Nulton TJ, Kim HW, Windle B, 
            Morgan IM, Munger K, White EA
  TITLE     PTPN14 degradation by high-risk human papillomavirus E7 limits 
            keratinocyte differentiation and contributes to HPV-mediated 
            oncogenesis.
  JOURNAL   Proc Natl Acad Sci U S A. 2019 Apr 2;116(14):7033-7042. doi: 
            10.1073/pnas.1819534116. Epub 2019 Mar 20.
  PUBMED    30894485
REFERENCE   2  (bases 1 to 6730)
  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"; doi: 
            "10.1073/pnas.1819534116"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2019-04-2- 2"; volume: "116"; issue: "14"; pages: "7033-7042"
COMMENT     Sequence Label: p7274 MSCV-P C-FlagHA 16 ∆DLYC
FEATURES             Location/Qualifiers
     source          1..6730
                     /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(846..870)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             902..925
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             941..967
                     /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             941..967
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        980..1479
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1015..1038)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1412..1431
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             1500..2099
                     /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(1500..1519)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1996..2016
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             2199..2713
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    2875..2891
                     /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(2899..2905,2906..2923,2924..2929))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2944..2965
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3082..3099)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3253..3841)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3333..3352)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4012..4803,4804..4872))
                     /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     4635..4654
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4873..4977)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5045..5063
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5101..5123)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5223..5242
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5436..5458
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5586..5605
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             5805..6321
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6385..6726
                     /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 tggagataca cctacattgc atgaatatat
      601 gttagatttg caaccagaga caacatatga gcaattaaat gacagctcag aggaggagga
      661 tgaaatagat ggtccggccg gacaagcaga accggacaga gcccattaca atattgtaac
      721 cttttgttgc aagtgtgact ctacgcttcg gttgtgcgta caaagcacac acgtagacat
      781 tcgtactttg gaagacctgt taatgggcac actaggaatt gtgtgcccca tctgttctca
      841 gaaaccacca actttcttgt acaaagtggt tgatctagag ggcccgcggt tcgaactcga
      901 ggactacaag gacgacgatg acaagtcggc cgctggagga tacccctacg acgtgcccga
      961 ctacgcctag aattctaccg ggtaggggag gcgcttttcc caaggcagtc tggagcatgc
     1021 gctttagcag ccccgctggg cacttggcgc tacacaagtg gcctctggcc tcgcacacat
     1081 tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt cgcgccacct
     1141 tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg
     1201 tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac cgctgagcaa
     1261 tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt cgctttctgg
     1321 gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct caggggcggg
     1381 gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct
     1441 gccgcgctgt tctcctcttc ctcatctccg ggcctttcga cctgcagccc aagcttacca
     1501 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac
     1561 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc
     1621 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     1681 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     1741 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     1801 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     1861 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     1921 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     1981 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     2041 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac
     2101 gcccgcccca cgacccgcag cgcccgaccg aaaggagcgc acgaccccat gcatcgataa
     2161 aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt
     2221 ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaatacat aactgagaat
     2281 agagaagttc agatcaaggt taggaacaga gagacagcag aatatgggcc aaacaggata
     2341 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg
     2401 tcccgccctc agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg
     2461 aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc
     2521 gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct
     2581 ccgatagact gcgtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc
     2641 gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc
     2701 gggggtcttt catgggtaac agtttcttga agttggagaa caacattctg agggtaggag
     2761 tcgaatatta agtaatcctg actcaattag ccactgtttt gaatccacat actccaatac
     2821 tcctgaaata gttcattatg gacagcgcag aaagagctgg ggagaattgt gaaattgtta
     2881 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     2941 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3001 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3061 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3121 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3181 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3241 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3301 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccttggaag
     3361 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3421 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3481 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3541 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3601 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3661 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3721 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3781 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3841 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3901 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3961 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4021 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4081 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4141 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4201 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4261 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4321 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4381 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4441 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4501 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     4561 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4621 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4681 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4741 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4801 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4861 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4921 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4981 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     5041 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     5101 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     5161 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     5221 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     5281 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     5341 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     5401 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     5461 ggcgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct
     5521 gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc
     5581 ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg
     5641 gccacgatgc gtccggcgta gaggcgatta gtccaatttg ttaaagacag gatatcagtg
     5701 gtccaggctc tagttttgac tcaacaatat caccagctga agcctataga gtacgagcca
     5761 tagataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc
     5821 tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa aatacataac
     5881 tgagaataga gaagttcaga tcaaggttag gaacagagag acagcagaat atgggccaaa
     5941 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca
     6001 gatgcggtcc cgccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa
     6061 ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg
     6121 ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca ctcggcgcgc
     6181 cagtcctccg atagactgcg tcgcccgggt acccgtattc ccaataaagc ctcttgctgt
     6241 ttgcatccga atcgtggact cgctgatcct tgggagggtc tcctcagatt gattgactgc
     6301 ccacctcggg ggtctttcat ttggaggttc caccgagatt tggagacccc tgcccaggga
     6361 ccaccgaccc ccccgccggg aggtaagctg gccagcggtc gtttcgtgtc tgtctctgtc
     6421 tttgtgcgtg tttgtgccgg catctaatgt ttgcgcctgc gtctgtacta gttagctaac
     6481 tagctctgta tctggcggac ccgtggtgga actgacgagt tctgaacacc cggccgcaac
     6541 cctgggagac gtcccaggga ctttgggggc cgtttttgtg gcccgacctg aggaagggag
     6601 tcgatgtgga atccgacccc gtcaggatat gtggttctgg taggagacga gaacctaaaa
     6661 cagttcccgc ctccgtctga atttttgctt tcggtttgga accgaagccg cgcgtcttgt
     6721 ctgctgcagc
//
