LOCUS Exported 6820 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION containing 1.3 units of the P-null HBV genome (subtype ayw). ACCESSION . VERSION . KEYWORDS HBV 1.3-mer P-null replicon SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6820) AUTHORS Wang H, Kim S, Ryu WS TITLE DDX3 DEAD-Box RNA helicase inhibits hepatitis B virus reverse transcription by incorporation into nucleocapsids. JOURNAL J Virol. 2009 Jun;83(11):5815-24. doi: 10.1128/JVI.00011-09. Epub 2009 Mar 18. PUBMED 19297497 REFERENCE 2 (bases 1 to 6820) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6820 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 842..1687 /codon_start=1 /product="middle (pre-S2/S) envelope protein of hepatitis B virus" /label=HBsAg (pre-S2/S) /note="hepatitis B surface antigen" /translation="MQWNSTTFHQTLQDPRVRGLYFPAGGSSSGTVNPVLTTASPLSSI FSRIGDPALNMENITSGFLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFLGGTTVCL GQNSQSPTSNHSPTSCPPTCPGYRWMCLRRFIIFLFILLLCLIFLLVLLDYQGMLPVCP LIPGSSTTSTGPCRTCMTTAQGTSMYPSCCCTKPSDGNCTCIPIPSSWAFGKFLWEWAS ARFSWLSLLVPFVQWFVGLSPTVWLSVIWMMWYWGPSLYSILSPFLPLLPIFFCLWVYI " primer_bind complement(2864..2883) /label=T7 /note="T7 promoter, forward primer" promoter complement(2865..2883) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2902..2918) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2902..2918) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2915..2937) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2926..2942 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(2950..2980) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2995..3016 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3133..3150) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3304..3892) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3384..3403) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4063..4923) /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 4686..4705 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4924..5028) /gene="bla" /label=AmpR promoter primer_bind 5096..5114 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(5152..5174) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 5274..5293 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5487..5509 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 5501..5518 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 5502..5518 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 5526..5544 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 5526..5543 /label=SP6 /note="SP6 promoter, forward primer" ORIGIN 1 ccggagacta ctgttgttag acgacgaggc aggtccccta gaagaagaac tccctcgcct 61 cgcagacgaa ggtctcaatc gccgcgtcgc agaagatctc aatctcggga atctcaatgt 121 tagtattcct tggactcata aggtggggaa ctttactggg ctttattctt ctactgtacc 181 tgtctttaat cctcattgga aaacaccatc ttttcctaat atacatttac accaagacat 241 tatcaaaaaa tgtgaacagt ttgtaggccc actcacagtt aatgagaaaa gaagattgca 301 attgattagc ctgccaggtt ttatccaaag gttaccaaat atttaccatt ggataagggt 361 attaaacctt attatccaga acatctagtt aatcattact tccaaactag acactattta 421 cacactctat ggaaggcggg tatattatat aagagagaaa caacacatag cgcctcattt 481 tgtgggtcac catattcttg ggaacaagat ctacagcatg gggcagaatc tttccaccag 541 caatcctctg ggattctttc ccgaccacca gttggatcca gccttcagag caaacaccgc 601 aaatccagat tgggacttca atcccaacaa ggacacctgg ccagacgcca acaaggtagg 661 agctggagca ttcgggctgg gtttcacccc accgcacgga ggccttttgg ggtggagccc 721 tcaggctcag ggcatactac aaactttgcc agcaaatccg cctcctgcct ccaccaatcg 781 ccagtcagga aggcagccta ccccgctgtc tccacctttg agaaacactc atcctcaggc 841 catgcagtgg aattccacaa ccttccacca aactctgcaa gatcccagag tgagaggcct 901 gtatttccct gctggtggct ccagttcagg aacagtaaac cctgttctga ctactgcctc 961 tcccttatcg tcaatcttct cgaggattgg ggaccctgcg ctgaacatgg agaacatcac 1021 atcaggattc ctaggacccc ttctcgtgtt acaggcgggg tttttcttgt tgacaagaat 1081 cctcacaata ccgcagagtc tagactcgtg gtggacttct ctcaattttc tagggggaac 1141 taccgtgtgt cttggccaaa attcgcagtc cccaacctcc aatcactcac caacctcttg 1201 tcctccaact tgtcctggtt atcgctggat gtgtctgcgg cgttttatca tcttcctctt 1261 catcctgctg ctatgcctca tcttcttgtt ggttcttctg gactatcaag gtatgttgcc 1321 cgtttgtcct ctaattccag gatcctcaac aaccagcacg ggaccatgcc ggacctgcat 1381 gactactgct caaggaacct ctatgtatcc ctcctgttgc tgtaccaaac cttcggacgg 1441 aaattgcacc tgtattccca tcccatcatc ctgggctttc ggaaaattcc tatgggagtg 1501 ggcctcagcc cgtttctcct ggctcagttt actagtgcca tttgttcagt ggttcgtagg 1561 gctttccccc actgtttggc tttcagttat atggatgatg tggtattggg ggccaagtct 1621 gtacagcatc ttgagtccct ttttaccgct gttaccaatt ttcttttgtc tttgggtata 1681 catttaaacc ctaacaaaac aaagagatgg ggttactctc taaattttat gggttatgtc 1741 attggatgtt atgggtcctt gccacaagaa cacatcatac aaaaaatcaa agaatgtttt 1801 agaaaacttc ctattaacag gcctattgat tggaaagtat gtcaacgaat tgtgggtctt 1861 ttgggttttg ctgccccttt tacacaatgt ggttatcctg cgttgatgcc tttgtatgca 1921 tgtattcaat ctaagcaggc tttcactttc tcgccaactt acaaggcctt tctgtgtaaa 1981 caatacctga acctttaccc cgttgcccgg caacggccag gtctgtgcca agtgtttgct 2041 gacgcaaccc ccactggctg gggcttggtc atgggccatc agcgcatgcg tggaaccttt 2101 tcggctcctc tgccgatcca tactgcggaa ctcctagccg cttgttttgc tcgcagcagg 2161 tctggagcaa acattatcgg gactgataac tctgttgtcc tatcccgcaa atatacatcg 2221 tttccatggc tgctaggctg tgctgccaac tggatcctgc gcgggacgtc ctttgtttac 2281 gtcccgtcgg cgctgaatcc tgcggacgac ccttctcggg gtcgcttggg actctctcgt 2341 ccccttctcc gtctgccgtt ccgaccgacc acggggcgca cctctcttta cgcggactcc 2401 ccgtctgtgc cttctcatct gccggaccgt gtgcacttcg cttcacctct gcacgtcgca 2461 tggagaccac cgtgaacgcc caccaaatat tgcccaaggt cttacataag aggactcttg 2521 gactctcagc aatgtcaacg accgaccttg aggcatactt caaagactgt ttgtttaaag 2581 actgggagga gttgggggag gagattaggt taaaggtctt tgtactagga ggctgtaggc 2641 ataaattggt ctgcgcacca gcaccatgca actttttcac ctctgcctaa tcatctcttg 2701 ttcatgtcct actgttcaag cctccaagct gtgccttggg tggctttggg gcatggacat 2761 cgacccttat aaagaattcg gagctactgt ggagttactc tcgtttttgc cttctgactt 2821 ctttccttca gtacgagatc ttctagatac cgcaagcttg tctccctata gtgagtcgta 2881 ttagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 2941 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 3001 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 3061 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 3121 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 3181 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 3241 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 3301 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 3361 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 3421 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 3481 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 3541 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 3601 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 3661 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 3721 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt 3781 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 3841 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 3901 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 3961 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 4021 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 4081 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 4141 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 4201 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 4261 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 4321 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 4381 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 4441 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 4501 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 4561 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 4621 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 4681 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 4741 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 4801 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 4861 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 4921 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 4981 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 5041 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 5101 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 5161 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 5221 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 5281 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 5341 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 5401 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 5461 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga 5521 attggattta ggtgacacta tagaatacga attcgagctc gtattcaatc taagcaggct 5581 ttcactttct cgccaactta caaggccttt ctgtgtaaac aatacctgaa cctttacccc 5641 gttgcccggc aacggccagg tctgtgccaa gtgtttgctg acgcaacccc cactggctgg 5701 ggcttggtca tgggccatca gcgcatgcgt ggaacctttt cggctcctct gccgatccat 5761 actgcggaac tcctagccgc ttgttttgct cgcagcaggt ctggagcaaa cattatcggg 5821 actgataact ctgttgtcct atcccgcaaa tatacatcgt ttccatggct gctaggctgt 5881 gctgccaact ggatcctgcg cgggacgtcc tttgtttacg tcccgtcggc gctgaatcct 5941 gcggacgacc cttctcgggg tcgcttggga ctctctcgtc cccttctccg tctgccgttc 6001 cgaccgacca cggggcgcac ctctctttac gcggactccc cgtctgtgcc ttctcatctg 6061 ccggaccgtg tgcacttcgc ttcacctctg cacgtcgcat ggagaccacc gtgaacgccc 6121 actaaatatt gcccaaggtc ttacataaga ggactcttgg actctcagca atgtcaacga 6181 ccgaccttga ggcatacttc aaagactgtt tgtttaaaga ctgggaggag ttgggggagg 6241 agattaggtt aaaggtcttt gtactaggag gctgtaggca taaattggtc tgcgcaccag 6301 caccatgcaa ctttttcacc tctgcctaat catctcttgt tcatgtccta ctgttcaagc 6361 ctccaagctg tgccttgggt ggctttgggg catggacatc gacccttata aagaatttgg 6421 agctactgtg gagttactct cgtttttgcc ttctgacttc tttccttcag tacgagatct 6481 tctagatacc gcctcagctc tgtatcggga agccttagag tctcctgagc attgttcacc 6541 tcaccatact gcactcaggc aagcaattct ttgctggggg gaactaatga ctctagctac 6601 ctgggtgggt gttaatttgg aagatccagc gtctagagac ctagtagtca gttatgtcaa 6661 cactaatatg ggcctaaagt tcaggcaact cttgtggttt cacatttctt gtctcacttt 6721 tggaagagaa acagttatag agtatttggt gtctttcgga gtgtggattc gcactcctcc 6781 agcttataga ccaccaaacg cccctatcct atcaacactt //