LOCUS       sequence-320724-        6913 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 6913)
  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 6913)
  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: p8220 MSCV-Neo C-HA 18E7 R84S
FEATURES             Location/Qualifiers
     source          1..6913
                     /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             460..486
                     /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             460..486
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        499..998
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(534..557)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     931..950
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             1019..1813
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             1019..1813
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1073..1092)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1683..1702
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             1892..2406
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    2568..2584
                     /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(2592..2598,2599..2616,2617..2622))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2637..2658
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2775..2792)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2946..3534)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3026..3045)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3705..4496,4497..4565))
                     /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     4328..4347
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4566..4670)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4738..4756
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4794..4816)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4916..4935
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5129..5151
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5279..5298
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             5498..6014
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6078..6419
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             6486..6902
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6832..6854
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6870..6886
                     /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 cgagtcggcc gctggaggat acccctacga cgtgcccgac
      481 tacgcctaga attctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg
      541 ctttagcagc cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt
      601 ccacatccac cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt
      661 ctactcctcc cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt
      721 gacaaatgga agtagcacgt ctcactagtc tcgtgcagat ggacagcacc gctgagcaat
      781 ggaagcgggt aggcctttgg ggcagcggcc aatagcagct ttgctccttc gctttctggg
      841 ctcagaggct gggaaggggt gggtccgggg gcgggctcag gggcgggctc aggggcgggg
      901 cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg
      961 ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac ctgcagccca agcttaccat
     1021 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     1081 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     1141 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     1201 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     1261 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     1321 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     1381 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     1441 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     1501 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     1561 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     1621 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     1681 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     1741 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     1801 cgagttcttc tgacgcccga ccgaaaggag cgcacgaccc catgcatcga taaaataaaa
     1861 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa
     1921 gctagcttaa gtaacgccat tttgcaaggc atggaaaata cataactgag aatagagaag
     1981 ttcagatcaa ggttaggaac agagagacag cagaatatgg gccaaacagg atatctgtgg
     2041 taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtcccgcc
     2101 ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac
     2161 cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg
     2221 ctccccgagc tcaataaaag agcccacaac ccctcactcg gcgcgccagt cctccgatag
     2281 actgcgtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt
     2341 ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc
     2401 tttcatgggt aacagtttct tgaagttgga gaacaacatt ctgagggtag gagtcgaata
     2461 ttaagtaatc ctgactcaat tagccactgt tttgaatcca catactccaa tactcctgaa
     2521 atagttcatt atggacagcg cagaaagagc tggggagaat tgtgaaattg ttatccgctc
     2581 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     2641 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     2701 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     2761 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     2821 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2881 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2941 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3001 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccttgg aagctccctc
     3061 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3121 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3181 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3241 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3301 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3361 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3421 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3481 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3541 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3601 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3661 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3721 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3781 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3841 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3901 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3961 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4021 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4081 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4141 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4201 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4261 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4321 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4381 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4441 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4501 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4561 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4621 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     4681 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     4741 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     4801 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     4861 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     4921 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
     4981 aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg
     5041 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     5101 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggcgcaa
     5161 ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca
     5221 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
     5281 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
     5341 tgcgtccggc gtagaggcga ttagtccaat ttgttaaaga caggatatca gtggtccagg
     5401 ctctagtttt gactcaacaa tatcaccagc tgaagcctat agagtacgag ccatagataa
     5461 aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt
     5521 ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaatacat aactgagaat
     5581 agagaagttc agatcaaggt taggaacaga gagacagcag aatatgggcc aaacaggata
     5641 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg
     5701 tcccgccctc agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg
     5761 aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc
     5821 gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct
     5881 ccgatagact gcgtcgcccg ggtacccgta ttcccaataa agcctcttgc tgtttgcatc
     5941 cgaatcgtgg actcgctgat ccttgggagg gtctcctcag attgattgac tgcccacctc
     6001 gggggtcttt catttggagg ttccaccgag atttggagac ccctgcccag ggaccaccga
     6061 cccccccgcc gggaggtaag ctggccagcg gtcgtttcgt gtctgtctct gtctttgtgc
     6121 gtgtttgtgc cggcatctaa tgtttgcgcc tgcgtctgta ctagttagct aactagctct
     6181 gtatctggcg gacccgtggt ggaactgacg agttctgaac acccggccgc aaccctggga
     6241 gacgtcccag ggactttggg ggccgttttt gtggcccgac ctgaggaagg gagtcgatgt
     6301 ggaatccgac cccgtcagga tatgtggttc tggtaggaga cgagaaccta aaacagttcc
     6361 cgcctccgtc tgaatttttg ctttcggttt ggaaccgaag ccgcgcgtct tgtctgctgc
     6421 agcgctgcag catcgttctg tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa
     6481 attagggcca gactgttacc actcccttaa gtttgacctt aggtcactgg aaagatgtcg
     6541 agcggatcgc tcacaaccag tcggtagatg tcaagaagag acgttgggtt accttctgct
     6601 ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga cggcaccttt aaccgagacc
     6661 tcatcaccca ggttaagatc aaggtctttt cacctggccc gcatggacac ccagaccagg
     6721 tcccctacat cgtgacctgg gaagccttgg cttttgaccc ccctccctgg gtcaagccct
     6781 ttgtacaccc taagcctccg cctcctcttc ctccatccgc cccgtctctc ccccttgaac
     6841 ctcctcgttc gaccccgcct cgatcctccc tttatccagc cctcactcct tctctaggcg
     6901 ccggaattag atc
//
