LOCUS       Exported                6534 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pGRB2.3, complete sequence.
ACCESSION   KF040397
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6534)
  AUTHORS   Zordan RE, Ren Y, Pan SJ, Rotondo G, Penas Ade L, Iluore J, Cormack 
            BP.
  TITLE     Expression Plasmids for Use in Candida glabrata
  JOURNAL   G3 (Bethesda) 3 (10), 1675-1686 (2013)
  PUBMED    23934995
REFERENCE   2  (bases 1 to 6534)
  AUTHORS   Zordan RE, Cormack BP.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-MAY-2013) Molecular Biology and Genetics, Johns 
            Hopkins School of Medicine, 725 N Wolfe St., Hunterian 609, 
            Baltimore, MD 21205, USA
REFERENCE   3  (bases 1 to 6534)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6534
                     /organism="Cloning vector pGRB2.3"
                     /mol_type="other DNA"
                     /db_xref="taxon:1386886"
     misc_feature    complement(66..664)
                     /label=Candida glabrata CEN/ARS
                     /note="Candida glabrata CEN/ARS"
     promoter        666..737
                     /gene="bla"
                     /label=AmpR promoter
     CDS             738..1598
                     /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"
     CDS             936..1598
                     /codon_start=1
                     /gene="bla"
                     /product="ampicillin-resistance protein"
                     /label=bla
                     /protein_id="AGT40319.1"
                     /translation="MSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHL
                     TDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEA
                     IPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFI
                     ADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     gene            936..1598
                     /gene="bla"
                     /label=bla
     rep_origin      1693..2375
                     /label=pBM1(ColE1) origin
                     /note="pBM1(ColE1) origin"
     rep_origin      1769..2357
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2645..2666
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2681..2711
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2719..2735
                     /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)."
     primer_bind     2743..2759
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2780..2798
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      2817..3367
                     /regulatory_class="promoter"
                     /note="Saccharomyces cerevisiae PGK1 promoter"
     primer_bind     3368..3384
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3404..4120
                     /codon_start=1
                     /gene="GFP"
                     /product="GFP"
                     /label=GFP
                     /protein_id="AGT40320.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     gene            3404..4120
                     /gene="GFP"
                     /label=GFP
     regulatory      4139..4536
                     /regulatory_class="terminator"
                     /note="region downstream of HIS3 in Candida glabrata BG2"
     promoter        complement(4551..4569)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4579..4595)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4736..5191
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(5322..6125)
                     /codon_start=1
                     /gene="URA3"
                     /product="orotidine 5-phosphate decarboxylase"
                     /label=URA3
                     /protein_id="AGT40321.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     gene            complement(5322..6125)
                     /gene="URA3"
                     /label=URA3
     promoter        complement(6126..6341)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter