LOCUS       sequence-132686-        5615 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To see the activity of HBV transcription using Luciferase assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5615)
  AUTHORS   Cha MY, Ryu DK, Jung HS, Chang HE, Ryu WS
  TITLE     Stimulation of hepatitis B virus genome replication by HBx is linked
            to both nuclear and cytoplasmic HBx expression.
  JOURNAL   J Gen Virol. 2009 Apr;90(Pt 4):978-86. doi: 10.1099/vir.0.009928-0. 
            Epub 2009 Mar 4.
  PUBMED    19264639
REFERENCE   2  (bases 1 to 5615)
  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 Gen 
            Virol."; date: "2009-04"; volume: "90(Pt 4):978-86. doi"; pages: " 
            10.1099/vir.0.009928-0. Epub 2009 Mar 4"
COMMENT     Sequence Label: pHBV-Luc
FEATURES             Location/Qualifiers
     source          1..5615
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      874..883
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             880..2532
                     /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(946..964)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2573..2694
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2584..2603)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2638..2657
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2942..2959)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3113..3701)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3193..3212)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3872..4663,4664..4732))
                     /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     4495..4514
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4733..4837)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4864..5319
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4951..4970)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5161..5182
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5450..5498
                     /note="synthetic polyadenylation signal"
     misc_feature    5512..5603
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5552..5571
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 cccgtattca atctaagcag gctttcactt tctcgccaac ttacaaggcc tttctgtgta
       61 aacaatacct gaacctttac cccgttgccc ggcaacggcc aggtctgtgc caagtgtttg
      121 ctgacgcaac ccccactggc tggggcttgg tcatgggcca tcagcgcatg cgtggaacct
      181 tttcggctcc tctgccgatc catactgcgg aactcctagc cgcttgtttt gctcgcagca
      241 ggtctggagc aaacattatc gggactgata actctgttgt cctatcccgc aaatatacat
      301 cgtttccatg gctgctaggc tgtgctgcta actggatcct gcgcgggacg tcctttgttt
      361 acgtcccgtc ggcgctgaat cctgcggacg acccttctcg gggtcgcttg ggactctctc
      421 gtccccttct ccgtctgccg ttccgaccga ccacggggcg caccactctt tacgcggact
      481 ccccgtctgt gccttctcat ctgccggacc gtgtgcactt cgcttcacct ctgcacgtcg
      541 catggagacc accgtgaacg cccactaaat attgctcaag gtcttacata agaggactct
      601 tggactctca gcaatgtcaa cgaccgacct tgaggcatac ttcaaagact gtttgtttaa
      661 agactgggag gagttggggg aggagattag gttaaaggtc tttgtactag gaggctgtag
      721 gcataaattg gtctgcgcac cagcaccatg caactttttc acctctgcct aatcatctct
      781 tgttcatgtc ctactgttca agcctccaag ctgtgccttg ggtggctttg gggcatggac
      841 acccaagctt ggcattccgg tactgttggt aaagccacca tggaagacgc caaaaacata
      901 aagaaaggcc cggcgccatt ctatccgctg gaagatggaa ccgctggaga gcaactgcat
      961 aaggctatga agagatacgc cctggttcct ggaacaattg cttttacaga tgcacatatc
     1021 gaggtggaca tcacttacgc tgagtacttc gaaatgtccg ttcggttggc agaagctatg
     1081 aaacgatatg ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa
     1141 ttctttatgc cggtgttggg cgcgttattt atcggagttg cagttgcgcc cgcgaacgac
     1201 atttataatg aacgtgaatt gctcaacagt atgggcattt cgcagcctac cgtggtgttc
     1261 gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa aaaagctccc aatcatccaa
     1321 aaaattatta tcatggattc taaaacggat taccagggat ttcagtcgat gtacacgttc
     1381 gtcacatctc atctacctcc cggttttaat gaatacgatt ttgtgccaga gtccttcgat
     1441 agggacaaga caattgcact gatcatgaac tcctctggat ctactggtct gcctaaaggt
     1501 gtcgctctgc ctcatagaac tgcctgcgtg agattctcgc atgccagaga tcctattttt
     1561 ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg ttccattcca tcacggtttt
     1621 ggaatgttta ctacactcgg atatttgata tgtggatttc gagtcgtctt aatgtataga
     1681 tttgaagaag agctgtttct gaggagcctt caggattaca agattcaaag tgcgctgctg
     1741 gtgccaaccc tattctcctt cttcgccaaa agcactctga ttgacaaata cgatttatct
     1801 aatttacacg aaattgcttc tggtggcgct cccctctcta aggaagtcgg ggaagcggtt
     1861 gccaagaggt tccatctgcc aggtatcagg caaggatatg ggctcactga gactacatca
     1921 gctattctga ttacacccga gggggatgat aaaccgggcg cggtcggtaa agttgttcca
     1981 ttttttgaag cgaaggttgt ggatctggat accgggaaaa cgctgggcgt taatcaaaga
     2041 ggcgaactgt gtgtgagagg tcctatgatt atgtccggtt atgtaaacaa tccggaagcg
     2101 accaacgcct tgattgacaa ggatggatgg ctacattctg gagacatagc ttactgggac
     2161 gaagacgaac acttcttcat cgttgaccgc ctgaagtctc tgattaagta caaaggctat
     2221 caggtggctc ccgctgaatt ggaatccatc ttgctccaac accccaacat cttcgacgca
     2281 ggtgtcgcag gtcttcccga cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg
     2341 gagcacggaa agacgatgac ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca
     2401 accgcgaaaa agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa aggtcttacc
     2461 ggaaaactcg acgcaagaaa aatcagagag atcctcataa aggccaagaa gggcggaaag
     2521 atcgccgtgt aattctagag tcggggcggc cggccgcttc gagcagacat gataagatac
     2581 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     2641 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     2701 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     2761 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cgtcgaccga tgcccttgag
     2821 agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact
     2881 tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt
     2941 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3001 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3061 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3121 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3181 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3241 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3301 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3361 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3421 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3481 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3541 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3601 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3661 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3721 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3781 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3841 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3901 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3961 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4021 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4081 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4141 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4201 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4261 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4321 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4381 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4441 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4501 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4561 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4621 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4681 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4741 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4801 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4861 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4921 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4981 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     5041 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     5101 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     5161 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     5221 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     5281 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttg ccattcgcca ttcaggctgc
     5341 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag cccaagctac
     5401 catgataagt aagtaatatt aaggtacggg aggtacttgg agcggccgca ataaaatatc
     5461 tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac taacatacgc
     5521 tctccatcaa aacaaaacga aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag
     5581 tgcaggtgcc agaacatttc tctatcgata ggtac
//
