LOCUS       sequence-46945-e        7053 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7053)
  AUTHORS   Johnson KM, Kines RC, Roberts JN, Lowy DR, Schiller JT, Day PM
  TITLE     Role of heparan sulfate in attachment to and infection of the murine
            female genital tract by human papillomavirus.
  JOURNAL   J Virol. 2009 Mar;83(5):2067-74. Epub 2008 Dec 10.
  PUBMED    19073722
REFERENCE   2  (bases 1 to 7053)
  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: "2009-03"; volume: "83(5)"; pages: "2067-74. Epub 2008 Dec 10"
COMMENT     Sequence Label: pCLucf
FEATURES             Location/Qualifiers
     source          1..7053
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1135..1144
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1141..2793
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     polyA_signal    2861..2982
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2872..2891)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2926..2945
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3168..3623
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3255..3274)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3465..3486
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3740..4097
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3740..3931
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3763..3783)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3948..4083
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      4122..4131
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4128..4130,4131..4133,4134..4847)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4128..4130,4131..4133,4134..4844)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4173..4194)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4434..4453)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4781..4802
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4902..4950
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4991..5010)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5110..5132
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5170..5188)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5256..5360
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5361..5429,5430..6221)
                     /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     complement(5579..5598)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6392..6980
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6881..6900
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc ctcgagaatt ctctagagct tggcaatccg gtactgttgg taaagccacc
     1141 atggaagatg ccaaaaacat taagaagggc ccagcgccat tctacccact cgaagacggg
     1201 accgccggcg agcagctgca caaagccatg aagcgctacg ccctggtgcc cggcaccatc
     1261 gcctttaccg acgcacatat cgaggtggac attacctacg ccgagtactt cgagatgagc
     1321 gttcggctgg cagaagctat gaagcgctat gggctgaata caaaccatcg gatcgtggtg
     1381 tgcagcgaga atagcttgca gttcttcatg cccgtgttgg gtgccctgtt catcggtgtg
     1441 gctgtggccc cagctaacga catctacaac gagcgcgagc tgctgaacag catgggcatc
     1501 agccagccca ccgtcgtatt cgtgagcaag aaagggctgc aaaagatcct caacgtgcaa
     1561 aagaagctac cgatcataca aaagatcatc atcatggata gcaagaccga ctaccagggc
     1621 ttccaaagca tgtacacctt cgtgacttcc catttgccac ccggcttcaa cgagtacgac
     1681 ttcgtgcccg agagcttcga ccgggacaaa accatcgccc tgatcatgaa cagtagtggc
     1741 agtaccggat tgcccaaggg cgtagcccta ccgcaccgca ccgcttgtgt ccgattcagt
     1801 catgcccgcg accccatctt cggcaaccag atcatccccg acaccgctat cctcagcgtg
     1861 gtgccatttc accacggctt cggcatgttc accacgctgg gctacttgat ctgcggcttt
     1921 cgggtcgtgc tcatgtaccg cttcgaggag gagctattct tgcgcagctt gcaagactat
     1981 aagattcaat ctgccctgct ggtgcccaca ctatttagct tcttcgctaa gagcactctc
     2041 atcgacaagt acgacctaag caacttgcac gagatcgcca gcggcggggc gccgctcagc
     2101 aaggaggtag gtgaggccgt ggccaaacgc ttccacctac caggcatccg ccagggctac
     2161 ggcctgacag aaacaaccag cgccattctg atcacccccg aaggggacga caagcctggc
     2221 gcagtaggca aggtggtgcc cttcttcgag gctaaggtgg tggacttgga caccggtaag
     2281 acactgggtg tgaaccagcg cggcgagctg tgcgtccgtg gccccatgat catgagcggc
     2341 tacgttaaca accccgaggc tacaaacgct ctcatcgaca aggacggctg gctgcacagc
     2401 ggcgacatcg cctactggga cgaggacgag cacttcttca tcgtggaccg gctgaagagc
     2461 ctgatcaaat acaagggcta ccaggtagcc ccagccgaac tggagagcat cctgctgcaa
     2521 caccccaaca tcttcgacgc cggggtcgcc ggcctgcccg acgacgatgc cggcgagctg
     2581 cccgccgcag tcgtcgtgct ggaacacggt aaaaccatga ccgagaagga gatcgtggac
     2641 tatgtggcca gccaggttac aaccgccaag aagctgcgcg gtggtgttgt gttcgtggac
     2701 gaggtgccta aaggactgac cggcaagttg gacgcccgca agatccgcga gattctcatt
     2761 aaggccaaga agggcggcaa gatcgccgtg taataattct agtggatccc ccgggcggcc
     2821 gcttcccttt agtgagggtt aatgcttcga gcagacatga taagatacat tgatgagttt
     2881 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     2941 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     3001 cattttatgt ttcaggttca gggggagatg tgggaggttt tttaaagcaa gtaaaacctc
     3061 tacaaatgtg gtaaaatccg ataaggatcg atccgggctg gcgtaatagc gaagaggccc
     3121 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggacg cgccctgtag
     3181 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3241 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3301 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3361 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3421 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3481 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     3541 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     3601 caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt
     3661 atttcacacc gcatacgcgg atctgcgcag caccatggcc tgaaataacc tctgaaagag
     3721 gaacttggtt aggtaccttc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     3781 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3841 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3901 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3961 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatacag
     4021 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4081 gcctaggctt ttgcaaaaag cttgattggg atccaccggt cgccaccatg gtgagcaagg
     4141 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     4201 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     4261 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     4321 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     4381 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     4441 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     4501 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     4561 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     4621 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     4681 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     4741 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     4801 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc ggcccgaaat
     4861 gaccgaccaa gcgacgccca acctgccatc acgatggccg caataaaata tctttatttt
     4921 cattacatct gtgtgttggt tttttgtgtg aatcgatagc gataaggatc cgcgtatggt
     4981 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     5041 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     5101 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     5161 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     5221 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     5281 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     5341 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     5401 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     5461 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     5521 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     5581 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     5641 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     5701 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     5761 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5821 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5881 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     5941 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     6001 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     6061 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     6121 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     6181 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     6241 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     6301 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6361 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6421 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6481 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     6541 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6601 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6661 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6721 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6781 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6841 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6901 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6961 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     7021 gctggccttt tgctcacatg gctcgacaga tct
//
