pGRB2.1 载体 (V005896#)

载体基本信息
载体名称 pGRB2.1 载体抗性 Ampicillin
载体长度 5299 bp 载体类型 Cloning vector
载体来源 Zordan RE, Ren Y, Pan SJ, Rotondo G, Penas Ade L, Iluore J, Cormack BP.

pGRB2.1 载体载体图谱

pGRB2.15299 bp6001200180024003000360042004800Candida glabrata CEN/ARSAmpR promoterAmpRpBM1(ColE1) originCAP binding sitelac promoterlac operatorM13 revT3 promoterSK primerKS primerregion downstream of HIS3 orf in Candida glabrata BG2T7 promoterM13 fwdf1 oriURA3URA3 promoter

pGRB2.1 载体载体序列

LOCUS       Exported                5299 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pGRB2.1, complete sequence.
ACCESSION   KF040395
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5299)
  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 5299)
  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 5299)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5299
                     /organism="Cloning vector pGRB2.1"
                     /mol_type="other DNA"
                     /db_xref="taxon:1386884"
     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="AGT40315.1"
                     /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"
     primer_bind     2835..2851
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2885..2901)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      2904..3301
                     /regulatory_class="terminator"
                     /note="region downstream of HIS3 orf in Candida glabrata 
                     BG2"
     promoter        complement(3316..3334)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3344..3360)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3501..3956
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(4087..4890)
                     /codon_start=1
                     /gene="URA3"
                     /product="orotidine 5-phosphate decarboxylase"
                     /label=URA3
                     /protein_id="AGT40316.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     gene            complement(4087..4890)
                     /gene="URA3"
                     /label=URA3
     promoter        complement(4891..5106)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter

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