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pPINKα-HC vector (V010238)

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

pPINKα-HC7898 bp30060090012001500180021002400270030003300360039004200450048005100540057006000630066006900720075007800AOX1 promoteralpha-factor secretion signalCYC1 terminatorPpADE2 HC promoterPpADE2PpTRP2 promoterPpTRP2M13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoterM13 fwd

pPINKα-HC vector Sequence

Copy Sequence

Download GeneBank File(.gb)

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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