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pPIC9K-His毕赤酵母表达质粒
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pPIC9K-His毕赤酵母表达质粒
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基本信息
产品名称:
pPIC9K-His毕赤酵母表达质粒
产品编号:
P1679
规格:
10μl质粒/300μl菌液
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LOCUS Exported 9294 bp ds-DNA circular SYN 26-NOV-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..9294
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 2..937
/gene="Pichia pastoris AOX1"
/label=AOX1 promoter
/note="inducible promoter, regulated by methanol"
CDS 949..1215
/codon_start=1
/gene="MF-alpha-1"
/product="N-terminal secretion signal from S. cerevisiae
alpha-factor"
/label=-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"
misc_feature 1216..1241
/label=MCS
misc_feature 1246..1263
/label=6×HIS
terminator 1339..1585
/gene="Pichia pastoris AOX1"
/label=AOX1 terminator
/note="transcription terminator for AOX1"
CDS complement(1998..4532)
/codon_start=1
/gene="Pichia pastoris HIS4"
/product="multifunctional enzyme, required for histidine
biosynthesis"
/label=PpHIS4
/note="auxotrophic marker for Pichia pastoris"
/translation="MTFPLLPAYASVAEFDNSLSLVGKAVFPYAADQLHNLIKFTQSTE
LQVNVQVESSVTEDQFEELIDNLLKLYNNGINEVILDLDLAERVVQRMIPGARVIYRTL
VDKVASLPANASIAVPFSSPLGDLKSFTNGGSRTVYAFSETAKLVDVTSTVASGIIPII
DARQLTTEYELSEDVKKFPVSEILLASLTTDRPDGLFTTLVADSSNYSLGLVYSSKKSI
PEAIRTQTGVYQSRRHGLWYKGATSGATQKLLGIELDCDGDCLKFVVEQTGVGFCHLER
TSCFGQSKGLRAMEATLWDRKSNAPEGSYTKRLFDDEVLLNAKIREEADELAEAKSKED
IAWECADLFYFALVRCAKYGVTLDEVERNLDMKSLKVTRRKGDAKPGYTKEQPKEESKP
KEVPSEGRIELCKIDVSKASSQEIEDALRRPIQKTEQIMELVKPIVDNVRQNGDKALLE
LTAKFDGVALKTPVLEAPFPEELMQLPDNVKRAIDLSIDNVRKFHEAQLTETLQVETCP
GVVCSRFARPIEKVGLYIPGGTAILPSTSLMLGVPAKVAGCKEIVFASPPKKDGTLTPE
VIYVAHKVGAKCIVLAGGAQAVAAMAYGTETVPKCDKIFGPGNQFVTAAKMMVQNDTSA
LCSIDMPAGPSEVLVIADKYADPDFVASDLLSQAEHGIDSQVILLAVDMTDKELARIED
AVHNQAVQLPRVEIVRKCIAHSTTLSVATYEQALEMSNQYAPEHLILQIENASSYVDQV
QHAGSVFVGAYSPESCGDYSSGTNHTLPTYGYARQYSGVNTATFQKFITSQDVTPEGLK
HIGQAVMDLAAVEGLDAHRNAVKVRMEKLGLI"
CDS complement(4946..5761)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
misc_feature 6140..6896
/label=AOX1 3' fragment
/note="region downstream of Pichia pastoris AOX1 gene"
misc_feature 7039..7179
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(7365..7953)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(8124..8984)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(8985..9089)
/gene="bla"
/label=AmpR promoter
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 aaagtcgcca 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 attcgaagga tccaaacgat gagatttcct
961 tcaattttta ctgcagtttt attcgcagca tcctccgcat tagctgctcc agtcaacact
1021 acaacagaag atgaaacggc acaaattccg gctgaagctg tcatcggtta ctcagattta
1081 gaaggggatt tcgatgttgc tgttttgcca ttttccaaca gcacaaataa cgggttattg
1141 tttataaata ctactattgc cagcattgct gctaaagaag aaggggtatc tctcgagaaa
1201 agagaggctg aagcttacgt agaattccct agggcggccg cgaatcatca tcaccatcac
1261 cattaattcg ccttagacat gactgttcct cagttcaagt tgggcactta cgagaagacc
1321 ggtcttgcta gattctaatc aagaggatgt cagaatgcca tttgcctgag agatgcaggc
1381 ttcatttttg atactttttt atttgtaacc tatatagtat aggatttttt ttgtcatttt
1441 gtttcttctc gtacgagctt gctcctgatc agcctatctc gcagctgatg aatatcttgt
1501 ggtaggggtt tgggaaaatc attcgagttt gatgtttttc ttggtatttc ccactcctct
1561 tcagagtaca gaagattaag tgagaagttc gtttgtgcaa gcttatcgat aagctttaat
1621 gcggtagttt atcacagtta aattgctaac gcagtcaggc accgtgtatg aaatctaaca
1681 atgcgctcat cgtcatcctc ggcaccgtca ccctggatgc tgtaggcata ggcttggtta
1741 tgccggtact gccgggcctc ttgcgggata tcgtccattc cgacagcatc gccagtcact
1801 atggcgtgct gctagcgcta tatgcgttga tgcaatttct atgcgcaccc gttctcggag
1861 cactgtccga ccgctttggc cgccgcccag tcctgctcgc ttcgctactt ggagccacta
1921 tcgactacgc gatcatggcg accacacccg tcctgtggat ctatcgaatc taaatgtaag
1981 ttaaaatctc taaataatta aataagtccc agtttctcca tacgaacctt aacagcattg
2041 cggtgagcat ctagaccttc aacagcagcc agatccatca ctgcttggcc aatatgtttc
2101 agtccctcag gagttacgtc ttgtgaagtg atgaacttct ggaaggttgc agtgttaact
2161 ccgctgtatt gacgggcata tccgtacgtt ggcaaagtgt ggttggtacc ggaggagtaa
2221 tctccacaac tctctggaga gtaggcacca acaaacacag atccagcgtg ttgtacttga
2281 tcaacataag aagaagcatt ctcgatttgc aggatcaagt gttcaggagc gtactgattg
2341 gacatttcca aagcctgctc gtaggttgca accgataggg ttgtagagtg tgcaatacac
2401 ttgcgtacaa tttcaaccct tggcaactgc acagcttggt tgtgaacagc atcttcaatt
2461 ctggcaagct ccttgtctgt catatcgaca gccaacagaa tcacctggga atcaatacca
2521 tgttcagctt gagacagaag gtctgaggca acgaaatctg gatcagcgta tttatcagca
2581 ataactagaa cttcagaagg cccagcaggc atgtcaatac tacacagggc tgatgtgtca
2641 ttttgaacca tcatcttggc agcagtaacg aactggtttc ctggaccaaa tattttgtca
2701 cacttaggaa cagtttctgt tccgtaagcc atagcagcta ctgcctgggc gcctcctgct
2761 agcacgatac acttagcacc aaccttgtgg gcaacgtaga tgacttctgg ggtaagggta
2821 ccatccttct taggtggaga tgcaaaaaca atttctttgc aaccagcaac tttggcagga
2881 acacccagca tcagggaagt ggaaggcaga attgcggttc caccaggaat atagaggcca
2941 actttctcaa taggtcttgc aaaacgagag cagactacac cagggcaagt ctcaacttgc
3001 aacgtctccg ttagttgagc ttcatggaat ttcctgacgt tatctataga gagatcaatg
3061 gctctcttaa cgttatctgg caattgcata agttcctctg ggaaaggagc ttctaacaca
3121 ggtgtcttca aagcgactcc atcaaacttg gcagttagtt ctaaaagggc tttgtcacca
3181 ttttgacgaa cattgtcgac aattggtttg actaattcca taatctgttc cgttttctgg
3241 ataggacgac gaagggcatc ttcaatttct tgtgaggagg ccttagaaac gtcaattttg
3301 cacaattcaa tacgaccttc agaagggact tctttaggtt tggattcttc tttaggttgt
3361 tccttggtgt atcctggctt ggcatctcct ttccttctag tgacctttag ggacttcata
3421 tccaggtttc tctccacctc gtccaacgtc acaccgtact tggcacatct aactaatgca
3481 aaataaaata agtcagcaca ttcccaggct atatcttcct tggatttagc ttctgcaagt
3541 tcatcagctt cctccctaat tttagcgttc aacaaaactt
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