LOCUS       sequence-323450-        6739 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HPV6 E7 R77S.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6739)
  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 6739)
  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: p8195 MSCV-P C-FlagHA 6E7 R77S
FEATURES             Location/Qualifiers
     source          1..6739
                     /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(855..879)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             911..934
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             950..976
                     /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             950..976
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        989..1488
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1024..1047)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1421..1440
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             1509..2108
                     /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(1509..1528)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2005..2025
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             2208..2722
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    2884..2900
                     /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(2908..2914,2915..2932,2933..2938))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2953..2974
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3091..3108)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3262..3850)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3342..3361)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4021..4812,4813..4881))
                     /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     4644..4663
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4882..4986)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5054..5072
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5110..5132)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5232..5251
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5445..5467
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5595..5614
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             5814..6330
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6394..6735
                     /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 gcatgcatgg aagacatgtt accctaaagg atattgtatt
      601 agacctgcaa cctccagacc ctgtagggtt acattgctat gagcaattag tagacagctc
      661 agaagatgag gtggacgaag tggacggaca agattcacaa cctttaaaac aacatttcca
      721 aatagtgacc tgttgctgtg gatgtgacag caacgttcga ctggttgtgc agtgtacaga
      781 aacagacatc agcgaagtgc aacagcttct gttgggaaca ctaaacatag tgtgtcccat
      841 ctgcgcaccg aagaccccaa ctttcttgta caaagtggtt gatctagagg gcccgcggtt
      901 cgaactcgag gactacaagg acgacgatga caagtcggcc gctggaggat acccctacga
      961 cgtgcccgac tacgcctaga attctaccgg gtaggggagg cgcttttccc aaggcagtct
     1021 ggagcatgcg ctttagcagc cccgctgggc acttggcgct acacaagtgg cctctggcct
     1081 cgcacacatt ccacatccac cggtaggcgc caaccggctc cgttctttgg tggccccttc
     1141 gcgccacctt ctactcctcc cctagtcagg aagttccccc ccgccccgca gctcgcgtcg
     1201 tgcaggacgt gacaaatgga agtagcacgt ctcactagtc tcgtgcagat ggacagcacc
     1261 gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc aatagcagct ttgctccttc
     1321 gctttctggg ctcagaggct gggaaggggt gggtccgggg gcgggctcag gggcgggctc
     1381 aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag
     1441 cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac ctgcagccca
     1501 agcttaccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca
     1561 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg
     1621 atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg
     1681 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca
     1741 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt
     1801 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc
     1861 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg
     1921 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg
     1981 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg
     2041 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg
     2101 gtgcctgacg cccgccccac gacccgcagc gcccgaccga aaggagcgca cgaccccatg
     2161 catcgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca
     2221 cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaatacata
     2281 actgagaata gagaagttca gatcaaggtt aggaacagag agacagcaga atatgggcca
     2341 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc
     2401 cagatgcggt cccgccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc
     2461 aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc
     2521 tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggcgc
     2581 gccagtcctc cgatagactg cgtcgcccgg gtacccgtgt atccaataaa ccctcttgca
     2641 gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta
     2701 cccgtcagcg ggggtctttc atgggtaaca gtttcttgaa gttggagaac aacattctga
     2761 gggtaggagt cgaatattaa gtaatcctga ctcaattagc cactgttttg aatccacata
     2821 ctccaatact cctgaaatag ttcattatgg acagcgcaga aagagctggg gagaattgtg
     2881 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     2941 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     3001 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3061 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3121 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3181 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3241 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3301 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3361 ccttggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3421 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3481 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3541 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3601 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3661 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3721 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3781 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3841 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3901 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3961 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4021 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4081 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4141 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4201 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4261 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4321 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4381 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4441 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4501 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4561 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4621 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4681 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4741 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4801 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4861 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4921 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4981 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     5041 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     5101 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     5161 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     5221 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     5281 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
     5341 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     5401 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     5461 taaaacgacg gcgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc
     5521 acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc
     5581 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
     5641 gtgatgccgg ccacgatgcg tccggcgtag aggcgattag tccaatttgt taaagacagg
     5701 atatcagtgg tccaggctct agttttgact caacaatatc accagctgaa gcctatagag
     5761 tacgagccat agataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa
     5821 gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa
     5881 atacataact gagaatagag aagttcagat caaggttagg aacagagaga cagcagaata
     5941 tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga
     6001 tggtccccag atgcggtccc gccctcagca gtttctagag aaccatcaga tgtttccagg
     6061 gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc
     6121 tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac
     6181 tcggcgcgcc agtcctccga tagactgcgt cgcccgggta cccgtattcc caataaagcc
     6241 tcttgctgtt tgcatccgaa tcgtggactc gctgatcctt gggagggtct cctcagattg
     6301 attgactgcc cacctcgggg gtctttcatt tggaggttcc accgagattt ggagacccct
     6361 gcccagggac caccgacccc cccgccggga ggtaagctgg ccagcggtcg tttcgtgtct
     6421 gtctctgtct ttgtgcgtgt ttgtgccggc atctaatgtt tgcgcctgcg tctgtactag
     6481 ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc
     6541 ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg cccgacctga
     6601 ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt aggagacgag
     6661 aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc
     6721 gcgtcttgtc tgctgcagc
//
