LOCUS       sequence-320721-        6742 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HPV16 E7 E10K.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6742)
  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 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"; 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: p7612 MSCV-P C-FlagHA 16E7 E10K
FEATURES             Location/Qualifiers
     source          1..6742
                     /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(368..392)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             424..447
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             463..489
                     /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             463..489
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        502..1001
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(537..560)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     934..953
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             1022..1621
                     /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(1022..1041)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1518..1538
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             1721..2235
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    2397..2413
                     /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(2421..2427,2428..2445,2446..2451))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2466..2487
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2604..2621)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2775..3363)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2855..2874)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3534..4325,4326..4394))
                     /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     4157..4176
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4395..4499)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4567..4585
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4623..4645)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4745..4764
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4958..4980
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5108..5127
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             5327..5843
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    5907..6248
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             6315..6731
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6661..6683
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6699..6715
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 taatacgact cactataggg agacccaagc tggctagtta agctatcaac aagtttgtac
       61 aaaaaagttg gcaccatgca tggagataca cctacattgc ataaatatat gttagatttg
      121 caaccagaga caactgatct ctactgttat gagcaattaa atgacagctc agaggaggag
      181 gatgaaatag atggtccagc tggacaagca gaaccggaca gagcccatta caatattgta
      241 accttttgtt gcaagtgtga ctctacgctt cggttgtgcg tacaaagcac acacgtagac
      301 attcgtactt tggaagacct gttaatgggc acactaggaa ttgtgtgccc catctgttct
      361 cagaaaccac caactttctt gtacaaagtg gttgatctag agggcccgcg gttcgaactc
      421 gaggactaca aggacgacga tgacaagtcg gccgctggag gataccccta cgacgtgccc
      481 gactacgcct agaattctac cgggtagggg aggcgctttt cccaaggcag tctggagcat
      541 gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg cctcgcacac
      601 attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac
      661 cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga
      721 cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc
      781 aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct
      841 gggctcagag gctgggaagg ggtgggtccg ggggcgggct caggggcggg ctcaggggcg
      901 gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa aagcgcacgt
      961 ctgccgcgct gttctcctct tcctcatctc cgggcctttc gacctgcagc ccaagcttac
     1021 catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt
     1081 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga
     1141 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     1201 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     1261 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     1321 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     1381 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     1441 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     1501 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     1561 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
     1621 acgcccgccc cacgacccgc agcgcccgac cgaaaggagc gcacgacccc atgcatcgat
     1681 aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag
     1741 gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaatac ataactgaga
     1801 atagagaagt tcagatcaag gttaggaaca gagagacagc agaatatggg ccaaacagga
     1861 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc
     1921 ggtcccgccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc
     1981 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
     2041 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc
     2101 ctccgataga ctgcgtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat
     2161 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca
     2221 gcgggggtct ttcatgggta acagtttctt gaagttggag aacaacattc tgagggtagg
     2281 agtcgaatat taagtaatcc tgactcaatt agccactgtt ttgaatccac atactccaat
     2341 actcctgaaa tagttcatta tggacagcgc agaaagagct ggggagaatt gtgaaattgt
     2401 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     2461 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     2521 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     2581 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     2641 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     2701 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2761 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2821 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccttgga
     2881 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2941 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3001 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3061 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3121 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3181 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3241 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3301 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3361 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3421 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     3481 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     3541 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     3601 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     3661 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3721 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3781 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3841 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     3901 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     3961 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4021 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4081 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4141 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     4201 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     4261 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     4321 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     4381 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     4441 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     4501 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     4561 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     4621 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     4681 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     4741 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     4801 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     4861 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     4921 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     4981 acggcgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc
     5041 ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt
     5101 ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc
     5161 cggccacgat gcgtccggcg tagaggcgat tagtccaatt tgttaaagac aggatatcag
     5221 tggtccaggc tctagttttg actcaacaat atcaccagct gaagcctata gagtacgagc
     5281 catagataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca
     5341 cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaatacata
     5401 actgagaata gagaagttca gatcaaggtt aggaacagag agacagcaga atatgggcca
     5461 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc
     5521 cagatgcggt cccgccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc
     5581 aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc
     5641 tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggcgc
     5701 gccagtcctc cgatagactg cgtcgcccgg gtacccgtat tcccaataaa gcctcttgct
     5761 gtttgcatcc gaatcgtgga ctcgctgatc cttgggaggg tctcctcaga ttgattgact
     5821 gcccacctcg ggggtctttc atttggaggt tccaccgaga tttggagacc cctgcccagg
     5881 gaccaccgac ccccccgccg ggaggtaagc tggccagcgg tcgtttcgtg tctgtctctg
     5941 tctttgtgcg tgtttgtgcc ggcatctaat gtttgcgcct gcgtctgtac tagttagcta
     6001 actagctctg tatctggcgg acccgtggtg gaactgacga gttctgaaca cccggccgca
     6061 accctgggag acgtcccagg gactttgggg gccgtttttg tggcccgacc tgaggaaggg
     6121 agtcgatgtg gaatccgacc ccgtcaggat atgtggttct ggtaggagac gagaacctaa
     6181 aacagttccc gcctccgtct gaatttttgc tttcggtttg gaaccgaagc cgcgcgtctt
     6241 gtctgctgca gcgctgcagc atcgttctgt gttgtctctg tctgactgtg tttctgtatt
     6301 tgtctgaaaa ttagggccag actgttacca ctcccttaag tttgacctta ggtcactgga
     6361 aagatgtcga gcggatcgct cacaaccagt cggtagatgt caagaagaga cgttgggtta
     6421 ccttctgctc tgcagaatgg ccaaccttta acgtcggatg gccgcgagac ggcaccttta
     6481 accgagacct catcacccag gttaagatca aggtcttttc acctggcccg catggacacc
     6541 cagaccaggt cccctacatc gtgacctggg aagccttggc ttttgacccc cctccctggg
     6601 tcaagccctt tgtacaccct aagcctccgc ctcctcttcc tccatccgcc ccgtctctcc
     6661 cccttgaacc tcctcgttcg accccgcctc gatcctccct ttatccagcc ctcactcctt
     6721 ctctaggcgc cggaattaga tc
//
