LOCUS       sequence-128516-        5204 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To see HBV X promoter(overlaped EnhanceI) activity for luciferase 
            assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5204)
  AUTHORS   Ko C, Lee S, Windisch MP, Ryu WS
  TITLE     DDX3 DEAD-box RNA helicase is a host factor that restricts hepatitis
            B virus replication at the transcriptional level.
  JOURNAL   J Virol. 2014 Dec;88(23):13689-98. doi: 10.1128/JVI.02035-14. Epub 
            2014 Sep 17.
  PUBMED    25231298
REFERENCE   2  (bases 1 to 5204)
  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"; doi: 
            "10.1128/JVI.02035-14"; journalName: "J Virol"; date: "2014-12"; 
            volume: "88"; issue: "23"; pages: "13689-98"
COMMENT     Sequence Label: pHBV-X/EnhI-Luc
FEATURES             Location/Qualifiers
     source          1..5204
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      468..477
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             474..2126
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(540..558)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2167..2288
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2178..2197)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2232..2251
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2536..2553)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2707..3295)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2787..2806)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3466..4257,4258..4326))
                     /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     4089..4108
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4327..4431)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4458..4913
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4545..4564)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4755..4776
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5044..5092
                     /note="synthetic polyadenylation signal"
     misc_feature    5106..5197
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5146..5165
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagtag aaaacttcct attaacaggc
       61 ctattgattg gaaagtatgt caacgaattg tgggtctttt gggttttgct gcccctttta
      121 cacaatgtgg ttatcctgcg ttgatgcctt tgtatgcatg tattcaatct aagcaggctt
      181 tcactttctc gccaacttac aaggcctttc tgtgtaaaca atacctgaac ctttaccccg
      241 ttgcccggca acggccaggt ctgtgccaag tgtttgctga cgcaaccccc actggctggg
      301 gcttggtcat gggccatcag cgcatgcgtg gaaccttttc ggctcctctg ccgatccata
      361 ctgcggaact cctagccgct tgttttgctc gcagcaggtc tggagcaaac attatcggga
      421 ctgataactc tgttgtccaa gcttggcatt ccggtactgt tggtaaagcc accatggaag
      481 acgccaaaaa cataaagaaa ggcccggcgc cattctatcc gctggaagat ggaaccgctg
      541 gagagcaact gcataaggct atgaagagat acgccctggt tcctggaaca attgctttta
      601 cagatgcaca tatcgaggtg gacatcactt acgctgagta cttcgaaatg tccgttcggt
      661 tggcagaagc tatgaaacga tatgggctga atacaaatca cagaatcgtc gtatgcagtg
      721 aaaactctct tcaattcttt atgccggtgt tgggcgcgtt atttatcgga gttgcagttg
      781 cgcccgcgaa cgacatttat aatgaacgtg aattgctcaa cagtatgggc atttcgcagc
      841 ctaccgtggt gttcgtttcc aaaaaggggt tgcaaaaaat tttgaacgtg caaaaaaagc
      901 tcccaatcat ccaaaaaatt attatcatgg attctaaaac ggattaccag ggatttcagt
      961 cgatgtacac gttcgtcaca tctcatctac ctcccggttt taatgaatac gattttgtgc
     1021 cagagtcctt cgatagggac aagacaattg cactgatcat gaactcctct ggatctactg
     1081 gtctgcctaa aggtgtcgct ctgcctcata gaactgcctg cgtgagattc tcgcatgcca
     1141 gagatcctat ttttggcaat caaatcattc cggatactgc gattttaagt gttgttccat
     1201 tccatcacgg ttttggaatg tttactacac tcggatattt gatatgtgga tttcgagtcg
     1261 tcttaatgta tagatttgaa gaagagctgt ttctgaggag ccttcaggat tacaagattc
     1321 aaagtgcgct gctggtgcca accctattct ccttcttcgc caaaagcact ctgattgaca
     1381 aatacgattt atctaattta cacgaaattg cttctggtgg cgctcccctc tctaaggaag
     1441 tcggggaagc ggttgccaag aggttccatc tgccaggtat caggcaagga tatgggctca
     1501 ctgagactac atcagctatt ctgattacac ccgaggggga tgataaaccg ggcgcggtcg
     1561 gtaaagttgt tccatttttt gaagcgaagg ttgtggatct ggataccggg aaaacgctgg
     1621 gcgttaatca aagaggcgaa ctgtgtgtga gaggtcctat gattatgtcc ggttatgtaa
     1681 acaatccgga agcgaccaac gccttgattg acaaggatgg atggctacat tctggagaca
     1741 tagcttactg ggacgaagac gaacacttct tcatcgttga ccgcctgaag tctctgatta
     1801 agtacaaagg ctatcaggtg gctcccgctg aattggaatc catcttgctc caacacccca
     1861 acatcttcga cgcaggtgtc gcaggtcttc ccgacgatga cgccggtgaa cttcccgccg
     1921 ccgttgttgt tttggagcac ggaaagacga tgacggaaaa agagatcgtg gattacgtcg
     1981 ccagtcaagt aacaaccgcg aaaaagttgc gcggaggagt tgtgtttgtg gacgaagtac
     2041 cgaaaggtct taccggaaaa ctcgacgcaa gaaaaatcag agagatcctc ataaaggcca
     2101 agaagggcgg aaagatcgcc gtgtaattct agagtcgggg cggccggccg cttcgagcag
     2161 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     2221 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     2281 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     2341 aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatccgtcga
     2401 ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact
     2461 atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca
     2521 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2581 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2641 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2701 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2761 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2821 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2881 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2941 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3001 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3061 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3121 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3181 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3241 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3301 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3361 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3421 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3481 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3541 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3601 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3661 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3721 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3781 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3841 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3901 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3961 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4021 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4081 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4141 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4201 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4261 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4321 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4381 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4441 ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4501 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4561 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4621 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4681 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4741 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4801 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4861 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttgccattc
     4921 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     4981 ccagcccaag ctaccatgat aagtaagtaa tattaaggta cgggaggtac ttggagcggc
     5041 cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgata
     5101 gtactaacat acgctctcca tcaaaacaaa acgaaacaaa acaaactagc aaaataggct
     5161 gtccccagtg caagtgcagg tgccagaaca tttctctatc gata
//
