LOCUS       Exported                6656 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  The 1.3xHBV-Luciferase plasmid to monitor HBV transcription. 1. 
            Luciferase inserted within Core ORF. 2. Luciferase is regulated by 
            EnhII - Core promoter. 3. The functional luciferase ATG is of Core.
ACCESSION   .
VERSION     .
KEYWORDS    1.3xWT HBV-Luciferase
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6656)
  AUTHORS   Doitsh G, Shaul Y
  TITLE     Enhancer I predominance in hepatitis B virus gene expression.
  JOURNAL   Mol Cell Biol. 2004 Feb;24(4):1799-808.
  PUBMED    14749394
REFERENCE   2  (bases 1 to 6656)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell 
            Biol."; date: "2004-02"; volume: "24(4)"; pages: "1799-808"
FEATURES             Location/Qualifiers
     source          1..6656
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             972..2621
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="EDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDAH
                     IEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAPA
                     NDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQSM
                     YTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHAR
                     DPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYKI
                     QSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYG
                     LTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMSG
                     YVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESILL
                     QHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVVF
                     VDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(1035..1053)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     promoter        complement(3981..3999)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3982..3999)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4017..4033)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4017..4033)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4030..4052)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4041..4057
                     /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(4065..4095)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4110..4131
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4248..4265)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4419..5007)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4499..4518)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5178..6038)
                     /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     5801..5820
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6039..6143)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6211..6229
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6267..6289)
                     /label=pGEX 3
                     /note="pGEX vectors, reverse primer"
     primer_bind     6389..6408
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6602..6624
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6616..6633
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6617..6633
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6640..6656,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(6640..6656,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"