To use pAbAi in a one-hybrid assay, clone one or more copies of a cis-acting element into the MCS. To facilitate recombination of the vector with the host’s ura3-52 locus, linearize the vector with either BbsI or BstBI, then introduce the linearized vector into competent yeast cells to integrate into the chromsome via homologous recombination between the vector’s URA3 gene and the ura3-52 locus of the yeast strain.
- 载体名称:
- pAbAi
- 载体抗性:
- Ampicillin
- 载体长度:
- 4894 bp
- 载体类型:
- Yeast Plasmids
- 复制子:
- ori
- 宿主:
- Yeast
- 载体来源:
- Clontech
- 拷贝数:
- High copy number
- 启动子:
- URA3
pAbAi 载体载体图谱
pAbAi 载体载体序列
LOCUS pAbAi. 4894 bp DNA circular SYN 01-JAN-1980 DEFINITION Yeast integrating vector for cloning a DNA bait sequence to be used in a one-hybrid screen. ACCESSION . VERSION . KEYWORDS pAbAi. SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4894) AUTHORS Clontech TITLE Direct Submission REFERENCE 2 (bases 1 to 4894) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" COMMENT Linearize with BstBI or BbsI to integrate at the URA3 locus. FEATURES Location/Qualifiers source 1..4894 /mol_type="other DNA" /organism="synthetic DNA construct" misc_feature 1..45 /label=MCS /note="MCS" /note="multiple cloning site" CDS 185..1387 /label=AurR /note="aureobasidin A-resistant mutant of S. cerevisiae inositol phosphorylceramide synthase (Aeed et al., 2009)" primer_bind complement(1645..1661) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(1669..1685) /label=lac operator /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(1693..1723) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(1738..1759) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(2047..2635) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(2809..3666) /label=AmpR /note="beta-lactamase" promoter complement(3667..3771) /label=AmpR promoter CDS complement(3878..4678) /label=URA3 /note="orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis" promoter complement(4679..4894) /label=URA3 promoter
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