LOCUS       sequence-200449-        7048 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human calreticulin fused to MmuPV1 E6, MmuPV1 E7 and 
            MmuPV1 L2 residues 11-210 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7048)
  AUTHORS   Jiang RT, Wang JW, Peng S, Huang TC, Wang C, Cannella F, Chang YN, 
            Viscidi RP, Best SRA, Hung CF, Roden RBS
  TITLE     Spontaneous and Vaccine-Induced Clearance of Mus Musculus 
            Papillomavirus 1 Infection.
  JOURNAL   J Virol. 2017 Jul 12;91(15). pii: e00699-17. doi: 
            10.1128/JVI.00699-17. Print 2017 Aug 1.
  PUBMED    28515303
REFERENCE   2  (bases 1 to 7048)
  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: "2017-07-12"; volume: "91(15). pii: e00699-17. doi"; pages: " 
            10.1128/JVI.00699-17. Print 2017 Aug 1"
COMMENT     Sequence Label: pNGVL4a-hCRTmE6mE7mL2
FEATURES             Location/Qualifiers
     source          1..7048
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        72..451
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        452..655
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     606..626
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     652..676
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          791..1060
                     /label=CMV intron
                     /note="modified intron A from human cytomegalovirus"
     primer_bind     complement(3717..3736)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3782..3837
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3836..3855)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3930..3947)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4101..4689)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4181..4200)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4992..5370)
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     primer_bind     5317..5336
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(5679..6494)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(5679..6494)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     6402..6421
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6486..6505
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     6772..6790
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6828..6850)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6950..6969
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 attgcatacg ttgtatccat atcataatat gtacatttat attggctcat gtccaacatt
       61 accgccatgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
      121 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
      181 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      241 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      301 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
      361 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
      421 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
      481 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
      541 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
      601 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgttt
      661 agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagaca
      721 ccgggaccga tccagcctcc gcggccggga acggtgcatt ggaacgcgga ttccccgtgc
      781 caagagtgac gtaagtaccg cctatagact ctataggcac acccctttgg ctcttatgca
      841 tgctatactg tttttggctt ggggcctata cacccccgct tccttatgct ataggtgatg
      901 gtatagctta gcctataggt gtgggttatt gaccattatt gaccactcca acggtggagg
      961 gcagtgtagt ctgagcagta ctcgttgctg ccgcgcgcgc caccagacat aatagctgac
     1021 agactaacag actgttcctt tccatgggtc ttttctgcag tcaccgtcgt cgacgccacc
     1081 atgctcctct cagtacctct tctcctcggt cttcttggac tcgctgttgc agaaccagca
     1141 gtatacttca aggagcagtt cctggacggt gacggatgga catcacgatg gatcgaatcc
     1201 aagcataagt ccgacttcgg taaattcgtc ctgagctccg gtaaattcta cggagacgag
     1261 gagaaggaca aggggctcca aacgtcccaa gacgcacgat tctacgcatt gtccgcttca
     1321 ttcgagccat tcagcaacaa gggacagact ctcgttgtac agttcacggt caagcatgag
     1381 cagaacatcg actgcggtgg aggttacgtt aagctgttcc caaactccct cgaccaaacg
     1441 gacatgcatg gagactctga gtacaacatc atgttcgggc ctgacatctg tggtccaggt
     1501 acgaagaagg tacacgtcat cttcaactac aaggggaaga acgtcctgat caacaaggac
     1561 atccgctgca aggacgacga gttcacgcac ctttacaccc tgatcgtccg accagacaac
     1621 acctacgagg taaagatcga caactcccag gtcgagtctg gttcacttga ggacgactgg
     1681 gatttccttc caccaaagaa gatcaaggac ccagacgctt ccaagcctga ggattgggat
     1741 gagcgagcta agatcgacga tccaactgac tccaagccag aagactggga taagccagaa
     1801 catatcccag acccagatgc taagaagcca gaagattggg acgaggagat ggacggtgag
     1861 tgggaaccac ctgttatcca aaaccctgag tacaagggcg agtggaagcc tagacagatc
     1921 gacaaccctg actacaaagg gacctggatc catcctgaga tcgacaatcc tgagtacagc
     1981 cctgaccctt ctatctacgc atacgacaac ttcggtgtgc tgggacttga cctctggcaa
     2041 gttaagtccg gaaccatctt cgacaacttc ctcatcacca acgacgaggc ttatgcagag
     2101 gaatttggga acgagacctg gggagtcaca aaggcagctg agaagcagat gaaggacaag
     2161 caggacgagg aacagagact taaggaggag gaagaggaca agaagagaaa agaggaggaa
     2221 gaagctgagg acaaagagga cgacgaggac aaagacgagg acgaggagga cgaggaagac
     2281 aaagaggagg acgaagagga ggatgtccct ggtcaagcaa aagacgagct tgaattcatg
     2341 gagatcggaa aagggtatac tctggaggaa gtgctcaggt attccaacaa ggacgtggtg
     2401 gatttccacc tgtcctgcgc tttttgctcc accaccatgg atcacaacga gaaagccaga
     2461 tttatccagg ctaaactgaa atgcgtcgtc agggattttg cctttaaagg ggcctgcatc
     2521 gtctgccgga ggcagctcgc gtgcaaagag aaactcctgc acaccagagt gaccggagaa
     2581 gcggatcttg tcgaatgcat ggccggaaaa aacatcgtgt ttgtgaccgt cagatgcgtg
     2641 acctgccttg cgttgttgac agcctctgaa aaactggatg ccaaagcctg cggattgcct
     2701 tttcacttgg tcagacacat gtggagggga tattgtggct tttgtaaacc cctgctgatg
     2761 caggggcctc tccctactat tgcggatatt gaaattcaaa atctggatag cctgctgggc
     2821 gtcggagaac ctgatctgcc tgatgtggga agcagttcac tgtcaccgga ttcattggga
     2881 gaagaagaag aactggaact ggaaaccatt gatgtggatc cctatcgcat taagaccacc
     2941 tgtttttgtt gtgataccgt gctgcggttt ataattgtca ccggcgatga tagcgtcaaa
     3001 gcctttgaaa gcctgctgct gcaggatctc agctttgtgt gtccccactg tgtggccagc
     3061 tatgtgaatc tgcggaatgg gaaaaggtct agaatgaaac gggatagcgc cagcaatctc
     3121 tatcggcagt gtcaggtgac tgggaattgt ccccccgatg tggtgaataa ggtggaaggg
     3181 aatacactgg ccgatcgcat actgaaagtg attagcagca ttgtgtatct gggcggcctg
     3241 gggattggca ctgggcgcgg gagcggagga acaactggct atggccccat aaatagtgcc
     3301 ggcggcaggg ttactgggac aggcactgtt atgaggccgg gcgttacagt agaacccatt
     3361 ggccccgggg atatagttac tgtagattct gtaggccccg gcgatagttc tttgattccc
     3421 ttgttggaag ttacaccgga tgttccgatt aatggcggcc ccgaagtgcc gtctagtggc
     3481 cccgatatta gtactgtgga tgtgacaagt tctattgatc ccatatctga tttgagtgtg
     3541 acaggcacaa caataagtaa tactgattca gccgtgattg atgtgcagcc ctctcccggc
     3601 ccgcggcgcg tgataattac acgctctgat tttaataatc ccagttatgt gtcagtggtg
     3661 catcccacac agggcttggg cgaatctggc ggcgtgtgag cggccgcgga tccagatctt
     3721 tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc
     3781 taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc
     3841 ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt
     3901 ttggcaacat atgcccattc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3961 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4021 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4081 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4141 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4201 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4261 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4321 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4381 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4441 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4501 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4561 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4621 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4681 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4741 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4801 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4861 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4921 gttgcctgac tcgggggggg gggcgctgag gtctgcctcg tgaagaaggt gttgctgact
     4981 cataccaggg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5041 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5101 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5161 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5221 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5281 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5341 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5401 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc tgaatcgccc catcatccag
     5461 ccagaaagtg agggagccac ggttgatgag agctttgttg taggtggacc agttggtgat
     5521 tttgaacttt tgctttgcca cggaacggtc tgcgttgtcg ggaagatgcg tgatctgatc
     5581 cttcaactca gcaaaagttc gatttattca acaaagccgc cgtcccgtca agtcagcgta
     5641 atgctctgcc agtgttacaa ccaattaacc aattctgatt agaaaaactc atcgagcatc
     5701 aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt
     5761 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
     5821 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
     5881 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa
     5941 agcttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
     6001 tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg
     6061 cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact
     6121 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct
     6181 gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
     6241 ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta
     6301 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
     6361 ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac
     6421 ccatataaat cagcatccat gttggaattt aatcgcggcc tcgagcaaga cgtttcccgt
     6481 tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt
     6541 catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga cacaacgtgg
     6601 ctttcccccc ccccccatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6661 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6721 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     6781 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     6841 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     6901 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     6961 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     7021 aaataccgca tcagattggc tattggcc
//
