pPICZαA
PVT4006 Packing 2ug
pPICZαA Information
Promoter: TEF1, AOX1, EM7
Replicon: pUC ori
Terminator: AOX1 TT
Plasmid classification: yeast series, Pichia pastoris expression vector
Plasmid size: 3593bp
Plasmid tagging: C-Myc, C-His
Prokaryotic resistance: Zeo
Selection marker: Zeo
Clonal strain: DH5 alpha
Culture conditions: 37 C, aerobic LB
Expression host: yeast cells
Induction mode: methanol induction
5'sequencing primers: AOX5:GACTGGTTCCAATTGACAAGC
3'sequencing primers: AOX3:GGCAAATGGCATTCTGACAT
pPICZαA Description
PPICZ alpha A, B and C are 3.6 KB Pichia pastoris protein secreting expression vectors. The expressed recombinant protein is a fusion protein containing an N-terminal polypeptide encoding the secretory signal of a-factor from Saccharomyces cerevisiae. The vector can express the target protein at a high level in Pichia pastoris induced by methanol, and can be used in any Pichia pastoris, including X33, GS115, SMD1168H, KM71H. PPICZ alpha A, B and C series carriers contain the following contents:
(1) Strict regulation of the promoter of AOX1 at the 5'-terminus induces the expression of any gene of interest (Ellis et al., 1985; Koutz et al., 1989; tschopp et al., 1987A).
(2) alpha factor secreting signal can secrete the target protein.
(3) Zeocin resistance genes can be used for screening in E. coli and Pichia pastoris (Baron et al., 1992; Drocourt et al., 1990).
(4) C- ends contain Myc and His tags, which can be used to detect and purify recombinant proteins.
(5) pPICZ A, B, C three reading frames make it possible to clone the gene into the vector without any frame-shifting mutation.
pPICZαA Multiple cloning site
pPICZαA Sequence
LOCUS Exported 3593 bp ds-DNA circular SYN 10-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 5
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3593)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Saturday, September 10, 2016 from SnapGene Viewer 3.1.4
FEATURES Location/Qualifiers
source 1..3593
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 2..940
/gene="Pichia pastoris AOX1"
/note="AOX1 promoter"
/note="inducible promoter, regulated by methanol"
CDS 941..1207
/codon_start=1
/gene="MF-alpha-1"
/product="N-terminal secretion signal from S. cerevisiae
alpha-factor"
/note="alpha-factor secretion signal"
/note="Cleavage by the Kex2 protease occurs after the
dibasic KR sequence. The EA dipeptides are then removed by
dipeptidyl aminopeptidase A."
/translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA"
CDS 1275..1304
/codon_start=1
/product="Myc (human c-Myc oncogene) epitope tag"
/note="Myc"
/translation="EQKLISEEDL"
CDS 1320..1337
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
terminator 1417..1663
/gene="Pichia pastoris AOX1"
/note="AOX1 terminator"
/note="transcription terminator for AOX1"
promoter 1678..2089
/gene="S. cerevisiae TEF1"
/note="TEF1 promoter"
/note="promoter for EF-1-alpha"
promoter 2097..2144
/note="EM7 promoter"
/note="synthetic bacterial promoter "
CDS 2163..2537
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/note="BleoR"
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
terminator 2603..2850
/gene="S. cerevisiae CYC1"
/note="CYC1 terminator"
/note="transcription terminator for CYC1"
rep_origin complement(2925..3513)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 agatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag
61 gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt
121 tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc
181 agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta
241 acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta
301 tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg
361 agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct
421 gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg
481 ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcggca taccgtttgt
541 cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct
601 ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct
661 ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact
721 gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat
781 atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt
841 actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga
901 caacttgaga agatcaaaaa acaactaatt attcgaaacg atgagatttc cttcaatttt
961 tactgctgtt ttattcgcag catcctccgc attagctgct ccagtcaaca ctacaacaga
1021 agatgaaacg gcacaaattc cggctgaagc tgtcatcggt tactcagatt tagaagggga
1081 tttcgatgtt gctgttttgc cattttccaa cagcacaaat aacgggttat tgtttataaa
1141 tactactatt gccagcattg ctgctaaaga agaaggggta tctctcgaga aaagagaggc
1201 tgaagctgaa ttcacgtggc ccagccggcc gtctcggatc ggtacctcga gccgcggcgg
1261 ccgccagctt tctagaacaa aaactcatct cagaagagga tctgaatagc gccgtcgacc
1321 atcatcatca tcatcattga gtttgtagcc ttagacatga ctgttcctca gttcaagttg
1381 ggcacttacg agaagaccgg tcttgctaga ttctaatcaa gaggatgtca gaatgccatt
1441 tgcctgagag atgcaggctt catttttgat acttttttat ttgtaaccta tatagtatag
1501 gatttttttt gtcattttgt ttcttctcgt acgagcttgc tcctgatcag cctatctcgc
1561 agctgatgaa tatcttgtgg taggggtttg ggaaaatcat tcgagtttga tgtttttctt
1621 ggtatttccc actcctcttc agagtacaga agattaagtg agaccttcgt ttgtgcggat
1681 cccccacaca ccatagcttc aaaatgtttc tactcctttt ttactcttcc agattttctc
1741 ggactccgcg catcgccgta ccacttcaaa acacccaagc acagcatact aaattttccc
1801 tctttcttcc tctagggtgt cgttaattac ccgtactaaa ggtttggaaa agaaaaaaga
1861 gaccgcctcg tttctttttc ttcgtcgaaa aaggcaataa aaatttttat cacgtttctt
1921 tttcttgaaa tttttttttt tagttttttt ctctttcagt gacctccatt gatatttaag
1981 ttaataaacg gtcttcaatt tctcaagttt cagtttcatt tttcttgttc tattacaact
2041 ttttttactt cttgttcatt agaaagaaag catagcaatc taatctaagg ggcggtgttg
2101 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
2161 ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg
2221 tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg
2281 gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg
2341 acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg
2401 aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc
2461 agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg
2521 ccgaggagca ggactgacac gtccgacggc ggcccacggg tcccaggcct cggagatccg
2581 tccccctttt cctttgtcga tatcatgtaa ttagttatgt cacgcttaca ttcacgccct
2641 ccccccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct
2701 atttattttt ttatagttat gttagtatta agaacgttat ttatatttca aatttttctt
2761 ttttttctgt acagacgcgt gtacgcatgt aacattatac tgaaaacctt gcttgagaag
2821 gttttgggac gctcgaaggc tttaatttgc aagctggaga ccaacatgtg agcaaaaggc
2881 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
2941 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
3001 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
3061 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa
3121 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
3181 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
3241 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
3301 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
3361 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
3421 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
3481 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
3541 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag atc
//
Caution:
- This product is FOR RESEARCH USE ONLY!
- The item is lyophilized form, Please take the powder plasmid by centrifugation at 5000rpm/min for 1min. Add 20μl ddH2O in to the tube of plasmid.Take 2ul plasmid into 100 μ L corresponding competent cell, centrifugal then full coating on plate.
- Shipping temperature is 2-8℃.
