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 Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
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"
This page is informational only.