LOCUS       sequence-197278-        7049 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 7049)
  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 7049)
  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..7049
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..456
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        457..660
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     611..631
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     657..681
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          796..1066
                     /label=CMV intron
                     /note="modified intron A from human cytomegalovirus"
     polyA_signal    3787..3841
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3840..3859)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3933..3950)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4104..4692)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4184..4203)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4997..5375)
                     /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     5322..5341
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(5684..6499)
                     /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(5684..6499)
                     /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     6407..6426
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6491..6510
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     6777..6795
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6834..6856)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6956..6975
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tggccattgc atacgttgta tccatatcat aatatgtaca tttatattgg ctcatgtcca
       61 acattaccgc catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
      121 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      181 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      241 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      301 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      361 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      421 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      481 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      541 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      601 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      661 cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga
      721 agacaccggg accgatccag cctccgcggc cgggaacggt gcattggaac gcggattccc
      781 cgtgccaaga gtgacgtaag taccgcctat agagtctata ggcccacccc cttggcttct
      841 tatgcatgct atactgtttt tggcttgggg tctatacacc cccgcttcct catgttatag
      901 gtgatggtat agcttagcct ataggtgtgg gttattgacc attattgacc actccaacgg
      961 tggagggcag tgtagtctga gcagtactcg ttgctgccgc gcgcgccacc agacataata
     1021 gctgacagac taacagactg ttcctttcca tgggtctttt ctgcagtcac cgtcgtcgac
     1081 gccaccatgc tcctctcagt acctcttctc ctcggtcttc ttggactcgc tgttgcagaa
     1141 ccagcagtat acttcaagga gcagttcctg gacggtgacg gatggacatc acgatggatc
     1201 gaatccaagc ataagtccga cttcggtaaa ttcgtcctga gctccggtaa attctacgga
     1261 gacgaggaga aggacaaggg gctccaaacg tcccaagacg cacgattcta cgcattgtcc
     1321 gcttcattcg agccattcag caacaaggga cagactctcg ttgtacagtt cacggtcaag
     1381 catgagcaga acatcgactg cggtggaggt tacgttaagc tgttcccaaa ctccctcgac
     1441 caaacggaca tgcatggaga ctctgagtac aacatcatgt tcgggcctga catctgtggt
     1501 ccaggtacga agaaggtaca cgtcatcttc aactacaagg ggaagaacgt cctgatcaac
     1561 aaggacatcc gctgcaagga cgacgagttc acgcaccttt acaccctgat cgtccgacca
     1621 gacaacacct acgaggtaaa gatcgacaac tcccaggtcg agtctggttc acttgaggac
     1681 gactgggatt tccttccacc aaagaagatc aaggacccag acgcttccaa gcctgaggat
     1741 tgggatgagc gagctaagat cgacgatcca actgactcca agccagaaga ctgggataag
     1801 ccagaacata tcccagaccc agatgctaag aagccagaag attgggacga ggagatggac
     1861 ggtgagtggg aaccacctgt tatccaaaac cctgagtaca agggcgagtg gaagcctaga
     1921 cagatcgaca accctgacta caaagggacc tggatccatc ctgagatcga caatcctgag
     1981 tacagccctg acccttctat ctacgcatac gacaacttcg gtgtgctggg acttgacctc
     2041 tggcaagtta agtccggaac catcttcgac aacttcctca tcaccaacga cgaggcttat
     2101 gcagaggaat ttgggaacga gacctgggga gtcacaaagg cagctgagaa gcagatgaag
     2161 gacaagcagg acgaggaaca gagacttaag gaggaggaag aggacaagaa gagaaaagag
     2221 gaggaagaag ctgaggacaa agaggacgac gaggacaaag acgaggacga ggaggacgag
     2281 gaagacaaag aggaggacga agaggaggat gtccctggtc aagcaaaaga cgagcttgaa
     2341 ttcatggaga tcggaaaagg gtatactctg gaggaagtgc tcaggtattc caacaaggac
     2401 gtggtggatt tccacctgtc ctgcgctttt tgctccacca ccatggatca caacgagaaa
     2461 gccagattta tccaggctaa actgaaatgc gtcgtcaggg attttgcctt taaaggggcc
     2521 tgcatcgtct gccggaggca gctcgcgtgc aaagagaaac tcctgcacac cagagtgacc
     2581 ggagaagcgg atcttgtcga atgcatggcc ggaaaaaaca tcgtgtttgt gaccgtcaga
     2641 tgcgtgacct gccttgcgtt gttgacagcc tctgaaaaac tggatgccaa agcctgcgga
     2701 ttgccttttc acttggtcag acacatgtgg aggggatatt gtggcttttg taaacccctg
     2761 ctgatgcagg ggcctctccc tactattgcg gatattgaaa ttcaaaatct ggatagcctg
     2821 ctgggcgtcg gagaacctga tctgcctgat gtgggaagca gttcactgtc accggattca
     2881 ttgggagaag aagaagaact ggaactggaa accattgatg tggatcccta tcgcattaag
     2941 accacctgtt tttgttgtga taccgtgctg cggtttataa ttgtcaccgg cgatgatagc
     3001 gtcaaagcct ttgaaagcct gctgctgcag gatctcagct ttgtgtgtcc ccactgtgtg
     3061 gccagctatg tgaatctgcg gaatgggaaa aggtctagaa tgaaacggga tagcgccagc
     3121 aatctctatc ggcagtgtca ggtgactggg aattgtcccc ccgatgtggt gaataaggtg
     3181 gaagggaata cactggccga tcgcatactg aaagtgatta gcagcattgt gtatctgggc
     3241 ggcctgggga ttggcactgg gcgcgggagc ggaggaacaa ctggctatgg ccccataaat
     3301 agtgccggcg gcagggttac tgggacaggc actgttatga ggccgggcgt tacagtagaa
     3361 cccattggcc ccggggatat agttactgta gattctgtag gccccggcga tagttctttg
     3421 attcccttgt tggaagttac accggatgtt ccgattaatg gcggccccga agtgccgtct
     3481 agtggccccg atattagtac tgtggatgtg acaagttcta ttgatcccat atctgatttg
     3541 agtgtgacag gcacaacaat aagtaatact gattcagccg tgattgatgt gcagccctct
     3601 cccggcccgc ggcgcgtgat aattacacgc tctgatttta ataatcccag ttatgtgtca
     3661 gtggtgcatc ccacacaggg cttgggcgaa tctggcggcg tgtgagcggc cgcggatcca
     3721 gatctttttc cctcgccaaa aattatgggg acatcatgaa gccccttgag catctgactt
     3781 ctggctaata aaggaaattt atttcattgc aatagtgtgt tggaattttt tgtgtctctc
     3841 actcggaagg acatatggga gggcaaatca tttaaaacat cagaatcagt atttggttta
     3901 gagtttggca acatatgcca ttcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3961 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4021 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4081 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4141 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4201 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4261 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     4321 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4381 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4441 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4501 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     4561 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4621 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4681 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4741 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4801 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4861 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4921 atagttgcct gactccgggg ggggggggcg ctgaggtctg cctcgtgaag aaggtgttgc
     4981 tgactcatac cagggcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5041 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5101 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5161 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5221 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5281 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5341 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5401 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacctgaat cgccccatca
     5461 tccagccaga aagtgaggga gccacggttg atgagagctt tgttgtaggt ggaccagttg
     5521 gtgattttga acttttgctt tgccacggaa cggtctgcgt tgtcgggaag atgcgtgatc
     5581 tgatccttca actcagcaaa agttcgattt attcaacaaa gccgccgtcc cgtcaagtca
     5641 gcgtaatgct ctgccagtgt tacaaccaat taaccaattc tgattagaaa aactcatcga
     5701 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     5761 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     5821 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     5881 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     5941 gcaaaagctt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     6001 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
     6061 atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     6121 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     6181 atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     6241 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
     6301 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     6361 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     6421 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctcgag caagacgttt
     6481 cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta
     6541 ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacaa
     6601 cgtggctttc cccccccccc cattattgaa gcatttatca gggttattgt ctcatgagcg
     6661 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6721 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     6781 ggcgtatcac gaggcccttt cgtcctcgcg cgtttcggtg atgacggtga aaacctctga
     6841 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     6901 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     6961 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
     7021 aggagaaaat accgcatcag attggctat
//
