LOCUS pPICZ B 3328 bp ds-DNA circular
DEFINITION Pichia pastoris vector for methanol-inducible intracellular
expression of a protein.
ACCESSION .
VERSION .
KEYWORDS pPICZ B
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT Linearize with SacI, PmeI, or BstXI for integration at the AOX1
locus.
FEATURES Location/Qualifiers
source 1..3328
/organism="synthetic DNA construct"
/lab_host="Pichia pastoris"
/mol_type="other DNA"
promoter 2..940
/gene="Pichia pastoris AOX1"
/note="AOX1 promoter"
/note="inducible promoter, regulated by methanol"
/note="color: #ffffff; direction: RIGHT"
misc_feature 932..1011
/note="MCS"
/note="multiple cloning site"
/note="color: #99ccff"
CDS 1010..1039
/codon_start=1
/product="Myc (human c-Myc oncogene) epitope tag"
/note="Myc"
/note="color: #cc99b2"
/translation="EQKLISEEDL"
CDS 1055..1072
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/note="color: #cc99b2"
/translation="HHHHHH"
terminator 1152..1398
/gene="Pichia pastoris AOX1"
/note="AOX1 terminator"
/note="transcription terminator for AOX1"
/note="color: #ffffff"
promoter 1438..1824
/gene="S. cerevisiae TEF1"
/note="TEF1 promoter"
/note="promoter for EF-1-alpha"
/note="color: #ffffff; direction: RIGHT"
promoter 1832..1879
/note="EM7 promoter"
/note="synthetic bacterial promoter?"
/note="color: #ffffff; direction: RIGHT"
CDS 1898..2272
/codon_start=1
/gene="Sh?ble from?Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/note="BleoR"
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/note="color: #ccffcc"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
terminator 2338..2585
/gene="S. cerevisiae CYC1"
/note="CYC1 terminator"
/note="transcription terminator for CYC1"
/note="color: #ffffff"
rep_origin complement(2660..3248)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
/note="color: #ffff00"
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 aggaattcac gtggcccagc
961 cggccgtctc ggatcggtac ctcgagccgc ggcggccgcc agctttctag aacaaaaact
1021 catctcagaa gaggatctga atagcgccgt cgaccatcat catcatcatc attgagtttg
1081 tagccttaga catgactgtt cctcagttca agttgggcac ttacgagaag accggtcttg
1141 ctagattcta atcaagagga tgtcagaatg ccatttgcct gagagatgca ggcttcattt
1201 ttgatacttt tttatttgta acctatatag tataggattt tttttgtcat tttgtttctt
1261 ctcgtacgag cttgctcctg atcagcctat ctcgcagctg atgaatatct tgtggtaggg
1321 gtttgggaaa atcattcgag tttgatgttt ttcttggtat ttcccactcc tcttcagagt
1381 acagaagatt aagtgagacc ttcgtttgtg cggatccccc acacaccata gcttcaaaat
1441 gtttctactc cttttttact cttccagatt ttctcggact ccgcgcatcg ccgtaccact
1501 tcaaaacacc caagcacagc atactaaatt ttccctcttt cttcctctag ggtgtcgtta
1561 attacccgta ctaaaggttt ggaaaagaaa aaagagaccg cctcgtttct ttttcttcgt
1621 cgaaaaaggc aataaaaatt tttatcacgt ttctttttct tgaaattttt ttttttagtt
1681 tttttctctt tcagtgacct ccattgatat ttaagttaat aaacggtctt caatttctca
1741 agtttcagtt tcatttttct tgttctatta caactttttt tacttcttgt tcattagaaa
1801 gaaagcatag caatctaatc taaggggcgg tgttgacaat taatcatcgg catagtatat
1861 cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt
1921 tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg
1981 gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct
2041 gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt
2101 gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga
2161 cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct
2221 gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtccg
2281 acggcggccc acgggtccca ggcctcggag atccgtcccc cttttccttt gtcgatatca
2341 tgtaattagt tatgtcacgc ttacattcac gccctccccc cacatccgct ctaaccgaaa
2401 aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata gttatgttag
2461 tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg
2521 catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg aaggctttaa
2581 tttgcaagct ggagaccaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
2641 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
2701 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
2761 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
2821 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt
2881 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
2941 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
3001 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
3061 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
3121 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
3181 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
3241 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
3301 cacgttaagg gattttggtc atgagatc
//