LOCUS       Exported                8299 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pHT3AG, complete sequence.
ACCESSION   GU046701
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8299)
  AUTHORS   Li Q, Yuan XE, Yin YP, Wang ZK.
  TITLE     Rapid determination of exogenous plasmid copy number and stability 
            using relative quantitative real-time PCR in recombinant 
            Brevibacillus brevis
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8299)
  AUTHORS   Li Q, Yuan XE, Yin YP, Wang ZK.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-SEP-2009) Bioengineering College of Chongqing 
            University, Genetic Engineering Research Center, Shazheng Street, 
            Chongqing, Chongqing 400030, China
REFERENCE   3  (bases 1 to 8299)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8299
                     /organism="Expression vector pHT3AG"
                     /lab_host="Brevibacillus brevis"
                     /mol_type="other DNA"
                     /label=derived from shuttle vector pHT304 and GFP gene
                     /note="derived from shuttle vector pHT304 and GFP gene"
                     /db_xref="taxon:689201"
     primer_bind     562..578
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1678..2394
                     /codon_start=1
                     /gene="gfp"
                     /product="GFP"
                     /label=gfp
                     /protein_id="ACY56286.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     gene            1678..2394
                     /gene="gfp"
                     /label=gfp
     primer_bind     complement(2419..2435)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2443..2459
                     /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(2467..2497)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2512..2533
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2821..3409)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3580..4440)
                     /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"