LOCUS       sequence-81735-e        9205 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HPV66 L1 and L2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9205)
  AUTHORS   Kwak K, Jiang R, Wang JW, Jagu S, Kirnbauer R, Roden RB
  TITLE     Impact of inhibitors and L2 antibodies upon the infectivity of 
            diverse alpha and beta human papillomavirus types.
  JOURNAL   PLoS One. 2014 May 9;9(5):e97232. doi: 10.1371/journal.pone.0097232.
            eCollection 2014.
  PUBMED    24816794
REFERENCE   2  (bases 1 to 9205)
  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: "PLoS One.";
            date: "2014-05-9"; volume: "9(5):e97232. doi"; pages: " 
            10.1371/journal.pone.0097232. eCollection 2014"
COMMENT     Sequence Label: pVITRO-HPV66 L1L2
FEATURES             Location/Qualifiers
     source          1..9205
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        51..242
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(74..94)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        256..1562
                     /label=mEF-1-alpha promoter
                     /note="strong constitutive promoter for mouse elongation 
                     factor EF-1-alpha"
     intron          583..1530
                     /label=mEF-1-alpha intron
                     /note="intron upstream of the start codon of mouse 
                     EF-1-alpha"
     regulatory      1582..1591
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3119..3240
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3130..3149)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3184..3203
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3428..4016
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3917..3936
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4096..4399
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4514..5819
                     /label=rEF-1-alpha promoter
                     /note="strong constitutive promoter for rat elongation 
                     factor EF-1-alpha"
     misc_feature    4856..5787
                     /label=rEF-1-alpha intron
                     /note="intron upstream of the start codon of rat 
                     EF-1-alpha"
     regulatory      5838..5847
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      5842..5851
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    7248..7692
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        7725..7771
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        7763..7781
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7791..8585
                     /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             7791..8585
                     /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(7845..7864)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8455..8474
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    8632..9204
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
ORIGIN
        1 cctgcagggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga
       61 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
      121 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
      181 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
      241 ccactagtgg agccgagagt aattcataca aaaggaggga tcgccttcgc aaggggagag
      301 cccagggacc gtccctaaat tctcacagac ccaaatccct gtagccgccc cacgacagcg
      361 cgaggagcat gcgctcaggg ctgagcgcgg ggagagcaga gcacacaagc tcatagaccc
      421 tggtcgtggg ggggaggacc ggggagctgg cgcggggcaa actgggaaag cggtgtcgtg
      481 tgctggctcc gccctcttcc cgagggtggg ggagaacggt atataagtgc ggcagtcgcc
      541 ttggacgttc tttttcgcaa cgggtttgcc gtcagaacgc aggtgagggg cgggtgtggc
      601 ttccgcgggc cgccgagctg gaggtcctgc tccgagcggg ccgggccccg ctgtcgtcgg
      661 cggggattag ctgcgagcat tcccgcttcg agttgcgggc ggcgcgggag gcagagtgcg
      721 aggcctagcg gcaaccccgt agcctcgcct cgtgtccggc ttgaggccta gcgtggtgtc
      781 cgcgccgccg ccgcgtgcta ctccggccgc actctggtct tttttttttt tgttgttgtt
      841 gccctgctgc cttcgattgc cgttcagcaa taggggctaa caaagggagg gtgcggggct
      901 tgctcgcccg gagcccggag aggtcatggt tggggaggaa tggagggaca ggagtggcgg
      961 ctggggcccg cccgccttcg gagcacatgt ccgacgccac ctggatgggg cgaggcctgg
     1021 ggtttttccc gaagcaacca ggctggggtt agcgtgccga ggccatgtgg ccccagcacc
     1081 cggcacgatc tggcttggcg gcgccgcgtt gccctgcctc cctaactagg gtgaggccat
     1141 cccgtccggc accagttgcg tgcgtggaaa gatggccgct cccgggccct gttgcaagga
     1201 gctcaaaatg gaggacgcgg cagcccggtg gagcgggcgg gtgagtcacc cacacaaagg
     1261 aagagggcct ggtccctcac cggctgctgc ttcctgtgac cccgtggtcc tatcggccgc
     1321 aatagtcacc tcgggctttt gagcacggct agtcgcggcg gggggagggg atgtaatggc
     1381 gttggagttt gttcacattt ggtgggtgga gactagtcag gccagcctgg cgctggaagt
     1441 catttttgga atttgtcccc ttgagttttg agcggagcta attctcgggc ttcttagcgg
     1501 ttcaaaggta tcttttaaac ccttttttag gtgttgtgaa aaccaccgct aattcaaagc
     1561 aaccggtgat atcaaagatc cgccaccatg gccatgtgga ggcccagcga caacaaggtg
     1621 tacctgcccc ccacccccgt gagcaaggtg gtggccaccg acacctacgt gaagaggacc
     1681 agcatcttct accacgccgg cagcagcagg ctgctggccg tgggccaccc ctactacagc
     1741 gtgagcaaga gcggcaccaa gaccaacatc cccaaggtga gcgcctacca gtacagggtg
     1801 ttcagggtga ggctccccga ccccaacaag ttcggcctgc ccgaccccag cttctacaac
     1861 cccgaccagg agaggctggt gtgggcctgc gtgggcctgg aggtgggcag gggccagccc
     1921 ctgggcgccg gcctgagcgg ccaccccctg ttcaacaggc tggacgacac cgaggtgagc
     1981 aacctggccg gcaacaacgt gatcgaggac agcagggaca acatcagcgt ggactgcaag
     2041 cagacccagc tgtgcatcgt gggctgcgcc cccgccctgg gcgagcactg gaccaagggc
     2101 gccgtgtgca agagcacccc cggcaacacc ggcgactgcc cccccctggc cctggtgaac
     2161 acccccatcg aggacggcga catggtggac accggcttcg gcgccatgga cttcaagctg
     2221 ctgcaggaga gcaaggccga ggtgcccctg gacatcgtgc agagcacctg caagtacccc
     2281 gactacctga agatgagcgc cgacgcctac ggcgacagca tgtggttcta cctgaggagg
     2341 gagcagctgt tcgccaggca ctacttcaac agggccggca acgtgggcga ggccatcccc
     2401 accgacctgt actggaaggg cggcaacggc agggaccccc cccccagcag cgtgtacgtg
     2461 gccaccccca gcggcagcat gatcaccagc gaggcccagc tgttcaacaa gccctactgg
     2521 ctgcagaggg cccagggcca caacaacggc atctgctggg gcaaccaggt gttcgtgacc
     2581 gtggtggaca ccaccaggag caccaacatg accatcaacg ccgccaagag caccctgacc
     2641 aagtacgacg ccagggagat caaccagtac ctgaggcacg tggaggagta cgagctgcag
     2701 ttcgtgttcc agctgtgcaa gatcaccctg accgccgagg tgatggccta cctgcacaac
     2761 atgaacaaca ccctgctgga cgactggaac atcggcctga gcccccccgt ggccaccagc
     2821 ctggaggaca agtacaggta catcaagagc accgccatca cctgccagag ggagcagccc
     2881 cccgccgaga agcaggaccc cctggccaag tacaagttct gggaggtgaa cctgcaggac
     2941 agcttcagcg ccgacctgga ccagttcccc ctgggcagga agttcctgat gcagctgggc
     3001 cccaggcccc ccaggcccaa ggccagcgtg agcgccagca agaggagggc cgcccccacc
     3061 agcagcagca gcagccccgc caagaggaag aagtaatagt ctagctggcc agacatgata
     3121 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     3181 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     3241 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     3301 taaagcaagt aaaacctcta caaatgtggt atggaaatgt taattaacta gccatgacca
     3361 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3421 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3481 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3541 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     3601 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3661 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3721 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3781 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     3841 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     3901 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     3961 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4021 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4081 taattaacct gcaggcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     4141 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     4201 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     4261 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     4321 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     4381 agtcatcgct attaccatga tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     4441 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     4501 actagtggag ccgagagtaa ttcatacaaa aggagggatc gccttcgcaa ggggagagcc
     4561 cagggaccgt ccctaaattc tcacagaccc aaatccctgt agccgcccca cgacagcgcg
     4621 aggagcatgc gcccagggct gagcgcgggt agatcagagc acacaagctc acagtccccg
     4681 gcggtggggg gaggggcgcg ctgagcgggg gccagggagc tggcgcgggg caaactggga
     4741 aagtggtgtc gtgtgctggc tccgccctct tcccgagggt gggggagaac ggtatataag
     4801 tgcggtagtc gccttggacg ttctttttcg caacgggttt gccgtcagaa cgcaggtgag
     4861 tggcgggtgt ggcttccgcg ggccccggag ctggagccct gctctgagcg ggccgggctg
     4921 atatgcgagt gtcgtccgca gggtttagct gtgagcattc ccacttcgag tggcgggcgg
     4981 tgcgggggtg agagtgcgag gcctagcggc aaccccgtag cctcgcctcg tgtccggctt
     5041 gaggcctagc gtggtgtccg ccgccgcgtg ccactccggc cgcactatgc gttttttgtc
     5101 cttgctgccc tcgattgcct tccagcagca tgggctaaca aagggagggt gtggggctca
     5161 ctcttaagga gcccatgaag cttacgttgg ataggaatgg aagggcagga ggggcgactg
     5221 gggcccgccc gccttcggag cacatgtccg acgccacctg gatggggcga ggcctgtggc
     5281 tttccgaagc aatcgggcgt gagtttagcc tacctgggcc atgtggccct agcactgggc
     5341 acggtctggc ctggcggtgc cgcgttccct tgcctcccaa caagggtgag gccgtcccgc
     5401 ccggcaccag ttgcttgcgc ggaaagatgg ccgctcccgg ggccctgttg caaggagctc
     5461 aaaatggagg acgcggcagc ccggtggagc gggcgggtga gtcacccaca caaaggaaga
     5521 gggccttgcc cctcgccggc cgctgcttcc tgtgaccccg tggtctatcg gccgcatagt
     5581 cacctcgggc ttctcttgag caccgctcgt cgcggcgggg ggaggggatc taatggcgtt
     5641 ggagtttgtt cacatttggt gggtggagac tagtcaggcc agcctggcgc tggaagtcat
     5701 tcttggaatt tgcccctttg agtttggagc gaggctaatt ctcaagcctc ttagcggttc
     5761 aaaggtattt tctaaacccg tttccaggtg ttgtgaaagc caccgctaat tcaaagcaat
     5821 ccggagtata cggatccgcc accatggtgg cccacagggc caccaggagg aagagggcca
     5881 gcgccaccca gctgtacaag acctgcaagc tgagcggcac ctgccccgag gacgtgatca
     5941 acaaggtgga gcagaagacc tgggccgaca ggattctgca gtggggcagc ctgttcacct
     6001 acttcggcgg cctgggcatc ggcacaggca gcgggagcgg cggcagggcc ggctacgtgc
     6061 ccctgggcag caggcccagc accatcgtgg acgtgacccc cgccaggccc cccatcgtgg
     6121 tggagagcgt gggccccacc gaccccagca tcgtgaccct ggtggaggag agcagcgtga
     6181 tcaacagcgg cgccggcgtg cccaacttca ccggcagcgg cggcttcgag gtgaccagca
     6241 gctccaccac cacccccgcc gtgctggaca tcacacccac cagcagcacc gtgcacgtga
     6301 gcagcaccac catcaccaac cccctgtaca tcgacccccc cgtgatcgag gccccccaga
     6361 ccggcgaggt gagcggcaac atcctgatca gcacccccac cagcggcatc cacagctacg
     6421 aggagatccc catgcagacc ttcgccatcc acggcaccgg caacgagccc atcagcagca
     6481 cccccatccc cggcttcagg aggctggccg cccccaggct gtacagcagg gccttccagc
     6541 aggtgagggt gaccgacccc gccttcctgg acaaccccac caccctgatc agcgccgaca
     6601 accccgtgtt cgagggcgcc gacaccaccc tgaccttcag ccccagcggc gtggcccccg
     6661 accccgactt catggacatc gtggccctgc acaggcccgc cttcaccacc aggaggaccg
     6721 gcgtgaggtt cagcaggctg ggcaagaagg ccaccatgca gaccaggagg ggcacccaga
     6781 tcggcgccag ggtgcactac tactacgaca tcagccccat cgcccaggcc gacgagatcg
     6841 agatgcagcc cctgctgagc accgacaaca gcttcgacgg cctgtacgac atctacgcca
     6901 acatcgacga cgaggccccc atcagcttca ggcagagcgg cgccaccccc agcgcccagc
     6961 tgcccatcaa gcccagcacc ctgagcttcg ccagcaacac cgccaacgtg accgcccccc
     7021 tgggcaacgt gtgggagacc cccttctaca gcggccccga catcgtgctg cccaccggcc
     7081 ccagcacctg gcccttcgtg ccccagagcc ccagcgacgt gacccacgac gtgtacatcc
     7141 agggcgccac cttcgccctg tggcccgtgt acttcttcaa gaggaggagg aggaagagga
     7201 ttccctactt cttcgccgac ggcgacgtgg ccgcctaata gtctaggagc aggtttcccc
     7261 aatgacacaa aacgtgcaac ttgaaactcc gcctggtctt tccaggtcta gaggggtaac
     7321 actttgtact gcgtttggct ccacgctcga tccactggcg agtgttagta acagcactgt
     7381 tgcttcgtag cggagcatga cggccgtggg aactcctcct tggtaacaag gacccacggg
     7441 gccaaaagcc acgcccacac gggcccgtca tgtgtgcaac cccagcacgg cgactttact
     7501 gcgaaaccca ctttaaagtg acattgaaac tggtacccac acactggtga caggctaagg
     7561 atgcccttca ggtaccccga ggtaacacgc gacactcggg atctgagaag gggactgggg
     7621 cttctataaa agcgctcggt ttaaaaagct tctatgcctg aataggtgac cggaggtcgg
     7681 cacctttcct ttgcaattac tgaccctatg aatacactga ctgtttgaca attaatcatc
     7741 ggcatagtat atcggcatag tataatacga ctcactatag gagggccacc atgattgaac
     7801 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     7861 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     7921 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     7981 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     8041 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     8101 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     8161 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     8221 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     8281 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     8341 tcgtcgtgac acatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     8401 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     8461 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     8521 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     8581 tctgagcggg actctggggt tcgaaatgac cgaccaagcg aattcgctag gattatccct
     8641 aatacctgcc accccactct taatcagtgg tggaagaacg gtctcagaac tgtttgtttc
     8701 aattggccat ttaagtttag tagtaaaaga ctggttaatg ataacaatgc atcgtaaaac
     8761 cttcagaagg aaaggagaat gttttgtgga ccactttggt tttctttttt gcgtgtggca
     8821 gttttaagtt attagttttt aaaatcagta ctttttaatg gaaacaactt gaccaaaaat
     8881 ttgtcacaga attttgagac ccattaaaaa agttaaatga gaaacctgtg tgttcctttg
     8941 gtcaacaccg agacatttag gtgaaagaca tctaattctg gttttacgaa tctggaaact
     9001 tcttgaaaat gtaattcttg agttaacact tctgggtgga gaatagggtt gttttccccc
     9061 cacataattg gaaggggaag gaatatcatt taaagctatg ggagggttgc tttgattaca
     9121 acactggaga gaaatgcagc atgttgctga ttgcctgtca ctaaaacagg ccaaaaactg
     9181 agtccttggg ttgcatagaa agctg
//
