LOCUS       Exported                6821 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  containing 1.3 units of the HBV genome (subtype ayw).
ACCESSION   .
VERSION     .
KEYWORDS    HBV 1.3-mer WT replicon
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6821)
  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 6821)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1128/JVI.00011-09"; journalName: "J Virol"; date: "2009-06"; 
            volume: "83"; issue: "11"; pages: "5815-24"
FEATURES             Location/Qualifiers
     source          1..6821
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             843..1688
                     /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(2865..2884)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2866..2884)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2903..2919)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2903..2919)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2916..2938)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2927..2943
                     /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(2951..2981)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2996..3017
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3134..3151)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3305..3893)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3385..3404)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4064..4924)
                     /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     4687..4706
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4925..5029)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5097..5115
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5153..5175)
                     /label=pGEX 3
                     /note="pGEX vectors, reverse primer"
     primer_bind     5275..5294
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5488..5510
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5502..5519
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5503..5519
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5527..5545
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5527..5544
                     /label=SP6
                     /note="SP6 promoter, forward primer"