LOCUS Exported 3151 bp ds-DNA circular SYN 13-SEP-2021
DEFINITION Pichia pastoris vector for constitutive expression of a secreted
protein. For other reading frames, use pGAPZ-alpha A or pGAPZ-alpha
C.
ACCESSION .
VERSION .
KEYWORDS pGAPZ-B
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3151)
AUTHORS Invitrogen (Life Technologies)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 3151)
AUTHORS .
TITLE Direct Submission
COMMENT Linearize within the GAP promoter for integration.
FEATURES Location/Qualifiers
source 1..3151
/organism="synthetic DNA construct"
/lab_host="Pichia pastoris"
/mol_type="other DNA"
promoter 7..483
/note="GAP promoter"
/note="GAP promoter;promoter for Pichia pastoris
glyceraldehyde-3-phosphate dehydrogenase"
CDS 493..759
/codon_start=1
/gene="MF-alpha-1"
/product="N-terminal secretion signal from S. cerevisiae
alpha-factor"
/note="MF-alpha-1"
/note="alpha-factor secretion signal;Cleavage by the Kex2
protease occurs after the dibasic KR sequence. The EA
dipeptides are then removed by dipeptidyl aminopeptidase
A."
/translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA"
misc_feature 758..832
/note="MCS"
/note="MCS;multiple cloning site"
CDS 831..860
/codon_start=1
/product="Myc (human c-Myc oncogene) epitope tag"
/note="Myc"
/translation="EQKLISEEDL"
CDS 876..893
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
terminator 972..1218
/gene="Pichia pastoris AOX1"
/note="Pichia pastoris AOX1 AOX1 terminator"
/note="AOX1 terminator;transcription terminator for AOX1"
promoter 1233..1644
/gene="S. cerevisiae TEF1"
/note="S. cerevisiae TEF1 TEF1 promoter"
/note="TEF1 promoter;promoter for EF-1-alpha"
promoter 1651..1698
/note="EM7 promoter"
/note="EM7 promoter;synthetic bacterial promoter"
CDS 1717..2091
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/note="Sh ble from Streptoalloteichus hindustanus"
/note="BleoR;confers resistance to bleomycin, phleomycin,
and Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
terminator 2157..2404
/gene="S. cerevisiae CYC1"
/note="S. cerevisiae CYC1 CYC1 terminator"
/note="CYC1 terminator;transcription terminator for CYC1"
rep_origin complement(2479..3067)
/direction=LEFT
/note="ori"
/note="ori;high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
LOCUS Exported 3151 bp ds-DNA circular SYN 13-SEP-2021
DEFINITION Pichia pastoris vector for constitutive expression of a secreted
protein. For other reading frames, use pGAPZ-alpha A or pGAPZ-alpha
C.
ACCESSION .
VERSION .
KEYWORDS pGAPZ-B
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3151)
AUTHORS Invitrogen (Life Technologies)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 3151)
AUTHORS .
TITLE Direct Submission
COMMENT Linearize within the GAP promoter for integration.
FEATURES Location/Qualifiers
source 1..3151
/organism="synthetic DNA construct"
/lab_host="Pichia pastoris"
/mol_type="other DNA"
promoter 7..483
/note="GAP promoter"
/note="GAP promoter;promoter for Pichia pastoris
glyceraldehyde-3-phosphate dehydrogenase"
CDS 493..759
/codon_start=1
/gene="MF-alpha-1"
/product="N-terminal secretion signal from S. cerevisiae
alpha-factor"
/note="MF-alpha-1"
/note="alpha-factor secretion signal;Cleavage by the Kex2
protease occurs after the dibasic KR sequence. The EA
dipeptides are then removed by dipeptidyl aminopeptidase
A."
/translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA"
misc_feature 758..832
/note="MCS"
/note="MCS;multiple cloning site"
CDS 831..860
/codon_start=1
/product="Myc (human c-Myc oncogene) epitope tag"
/note="Myc"
/translation="EQKLISEEDL"
CDS 876..893
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
terminator 972..1218
/gene="Pichia pastoris AOX1"
/note="Pichia pastoris AOX1 AOX1 terminator"
/note="AOX1 terminator;transcription terminator for AOX1"
promoter 1233..1644
/gene="S. cerevisiae TEF1"
/note="S. cerevisiae TEF1 TEF1 promoter"
/note="TEF1 promoter;promoter for EF-1-alpha"
promoter 1651..1698
/note="EM7 promoter"
/note="EM7 promoter;synthetic bacterial promoter"
CDS 1717..2091
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/note="Sh ble from Streptoalloteichus hindustanus"
/note="BleoR;confers resistance to bleomycin, phleomycin,
and Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
terminator 2157..2404
/gene="S. cerevisiae CYC1"
/note="S. cerevisiae CYC1 CYC1 terminator"
/note="CYC1 terminator;transcription terminator for CYC1"
rep_origin complement(2479..3067)
/direction=LEFT
/note="ori"
/note="ori;high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"