Basic Vector Information
The PichiaPink Secreted Vector Kit contains the pPINKα-HC vector for use with the PichiaPink Yeast Expression System. The pPINKα-HC vector is built around the ADE2 gene for complementing the complementing the ade2 deletion in the PichiaPink strains. The pPINKα-HC vector has the α-mating factor secretion signal sequence so that all proteins are secreted. You can order the PichiaPink Secreted Vector Kit to refill your PichiaPink Yeast Expression System Kit.
- Vector Name:
- pPINKα-HC
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7898 bp
- Type:
- Yeast Plasmids
- Replication origin:
- ori
- Source/Author:
- Invitrogen (Life Technologies)
- Selection Marker:
- ADE2
- Copy Number:
- High copy number
- Promoter:
- AOX1
- Cloning Method:
- Enzyme digestion and ligation
- 5' Primer:
- 5´ AOX1:5´-GACTGGTTCCAATTGACAAGC-3´ alpha-Factor-F: 5´-TACTATTGCCAGCATTGCTGC-3´
- 3' Primer:
- 3´ AOX1:5´-GCAAATGGCATTCTGACATCC-3´
pPINKα-HC vector Map
pPINKα-HC vector Sequence
LOCUS pPink-alpha-HC. 7898 bp DNA circular SYN 13-SEP-2021
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS pPink-alpha-HC
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7898)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 7898)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7898
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 2..940
/label=AOX1 promoter
/note="inducible promoter, regulated by methanol"
CDS 941..1195
/label=alpha-factor secretion signal
/note="N-terminal secretion signal from S. cerevisiae
alpha-factor"
terminator 1248..1497
/label=CYC1 terminator
/note="transcription terminator for CYC1"
promoter 1517..1529
/gene="Pichia pastoris ADE2"
/label=Pichia pastoris ADE2 promoter
/note="PpADE2 HC promoter"
/note="weak 13-bp promoter fragment that requires high-copy
integration for efficient expression"
CDS 1530..3218
/label=PpADE2
/note="phosphoribosylaminoimidazole carboxylase from Pichia
pastoris"
promoter 3448..3597
/label=PpTRP2 promoter
/note="promoter for Pichia pastoris TRP2"
CDS 3598..5211
/label=PpTRP2
/note="anthranilate synthase from Pichia pastoris"
primer_bind complement(5276..5292)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(5300..5316)
/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(5324..5354)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(5369..5390)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5678..6266)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6440..7297)
/label=AmpR
/note="beta-lactamase"
promoter complement(7298..7402)
/label=AmpR promoter
primer_bind 7876..7892
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
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