LOCUS pPinkα-HC 7898 bp ds-DNA circular SYN 04-二月-2013
DEFINITION Pichia pastoris integrating vector for high-copy expression of a
secreted protein in the PichiaPink(TM) system.
ACCESSION .
VERSION .
KEYWORDS pPINK(alpha)-HC
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 within PpTRP2 for integration.
FEATURES Location/Qualifiers
source 1..7898
/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"
CDS 941..1195
/codon_start=1
/gene="MF-alpha-1"
/product="N-terminal secretion signal from S. cerevisiae
alpha-factor (modified)"
/note="alpha-factor secretion signal"
/note="Cleavage by the Kex2 protease occurs after the
dibasic KR sequence."
/note="This feature has 2 segments:
???1:??941?..??997?/?#993366?/?presequence
???2:??998?..?1195?/?#993366?/?pro?region
Cleavage site after base 997"
/translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKR"
misc_feature 1196..1222
/note="MCS"
/note="multiple cloning site"
/note="color: #99ccff"
terminator 1248..1497
/gene="S. cerevisiae CYC1"
/note="CYC1 terminator"
/note="transcription terminator for CYC1"
/note="color: #ffffff"
promoter 1517..1529
/gene="Pichia pastoris ADE2"
/note="PpADE2 HC promoter"
/note="weak 13-bp promoter fragment that requires high-copy
integration for efficient expression"
/note="color: #ffffff; direction: RIGHT"
CDS 1530..3221
/codon_start=1
/gene="Pichia pastoris ADE2"
/product="phosphoribosylaminoimidazole carboxylase from
Pichia pastoris"
/note="PpADE2"
/note="essential for the biosynthesis of purine
nucleotides, including adenine"
/note="color: #ff7f50"
promoter 3448..3597
/gene="Pichia pastoris TRP2"
/note="PpTRP2 promoter"
/note="promoter for Pichia pastoris TRP2"
/note="color: #ffffff; direction: RIGHT"
CDS 3598..5214
/codon_start=1
/gene="Pichia pastoris TRP2"
/product="anthranilate synthase from Pichia pastoris"
/note="PpTRP2"
/note="catalyzes the initial step of tryptophan
biosynthesis"
/note="color: #ff7f50"
rep_origin complement(5678..6266)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
/note="color: #ffff00"
CDS complement(6437..7297)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
promoter complement(7298..7402)
/gene="bla"
/note="AmpR promoter"
/note="color: #ffffff; direction: LEFT"
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 aaaggcctca
1201 ggtaccggcc ggccatttaa atacaggccc cttttccttt gtcgatatca tgtaattagt
1261 tatgtcacgc ttacattcac gccctcctcc cacatccgct ctaaccgaaa aggaaggagt
1321 tagacaacct gaagtctagg tccctattta ttttttttaa tagttatgtt agtattaaga
1381 acgttattta tatttcaaat ttttcttttt tttctgtaca aacgcgtgta cgcatgtaac
1441 attatactga aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcaag
1501 ctggatccgc ggccgccttc caaactctca tggattctca ggtaataggt attctaggag
1561 gaggccagct aggccgaatg attgttgagg ccgctagcag gctcaatatc aagaccgtga
1621 ttcttgatga tggtttttca cctgctaagc acattaatgc tgcgcaagac cacatcgacg
1681 gatcattcaa agatgaggag gctatcgcca agttagctgc caaatgtgat gttctcactg
1741 tagagattga gcatgtcaac acagatgctc taaagagagt tcaagacaga actggaatca
1801 agatatatcc tttaccagag acaatcgaac taatcaagga taagtacttg caaaaggaac
1861 atttgatcaa gcacaacatt tcggtgacaa agtctcaggg tatagaatct aatgaaaagg
1921 cgctgctttt gtttggagaa gagaatggat ttccatatct gttgaagtcc cggactatgg
1981 cttatgatgg aagaggcaat tttgtagtgg agtctaaaga ggacatcagt aaggcattag
2041 agttcttgaa agatcgtcca ttgtatgccg agaagtttgc tccttttgtt aaagaattag
2101 cggtaatggt tgtgagatca ctggaaggcg aagtattctc ctacccaacc gtagaaactg
2161 tgcacaagga caatatctgt catattgtgt atgctccggc cagagttaat gacaccatcc
2221 aaaagaaagc tcaaatatta gctgaaaaca ctgtgaagac tttcccaggc gctggaatct
2281 tcggagttga gatgttccta ttgtctgatg gagaacttct tgtaaatgag attgctccaa
2341 ggccccacaa ttctggtcac tatacaatcg atgcatgtgt aacatctcag ttcgaagcac
2401 atgtaagagc cataactggt ctgccaatgc cactagattt caccaaacta tctacttcca
2461 acaccaacgc tattatgctc aatgttttgg gtgctgaaaa atctcacggg gaattagagt
2521 tttgtagaag agccttagaa acacccggtg cttctgtata tctgtacgga aagaccaccc
2581 gattggctcg taagatgggt catatcaaca taataggatc ttccatgttg gaagcagaac
2641 aaaagttaga gtacattcta gaagaatcaa cccacttacc atccagtact gtatcagctg
2701 acactaaacc gttggttgga gttatcatgg gttcagactc tgatctacct gtgatttcga
2761 aaggttgcga tattttaaaa cagtttggtg ttccattcga agttactatt gtctctgctc
2821 atagaacacc acagagaatg accagatatg cctttgaagc cgctagtaga ggtatcaagg
2881 ctatcattgc aggtgctggt ggtgctgctc atcttccagg aatggttgct gccatgactc
2941 cgttgccagt cattggtgtt cctgtcaagg gctctacgtt ggatggtgta gactcgctac
3001 actcgattgt ccaaatgcct agaggtgttc ctgtggctac ggttgctatc aacaacgcca
3061 ccaatgccgc tctgttggcc atcaggattt taggtacaat tgaccacaaa tggcaaaagg
3121 aaatgtccaa gtatatgaat gcaatggaga ccgaagtgtt ggggaaggca tccaacttgg
3181 aatctgaagg gtatgaatcc tatttgaaga atcgtctttg aatttagtat tgttttttaa
3241 tagatgtata tataatagta cacgtaactt atctattcca ttcataattt tattttaaag
3301 gttcggtaga aatttgtcct ccaaaaagtt ggttagagcc tggcagtttt gataggcatt
3361 attatagatt gggtaatatt taccctgcac ctggaggaac tttgcaaaga gcctcatgtg
3421 cggcgcgcca ggccataatg gccaaacggt ttctcaatta ctatatacta ctaaccattt
3481 acctgtagcg tatttctttt ccctcttcgc gaaagctcaa gggcatcttc ttgactcatg
3541 aaaaatatct ggatttcttc tgacagatca tcacccttga gcccaactct ctagcctatg
3601 agtgtaagtg atagtcatct tgcaacagat tattttggaa cgcaactaac aaagcagata
3661 cacccttcag cagaatcctt tctggatatt gtgaagaatg atcgccaaag tcacagtcct
3721 gagacagttc ctaatcttta ccccatttac aagttcatcc aatcagactt cttaacgcct
3781 catctggctt atatcaagct taccaacagt tcagaaactc ccagtccaag tttcttgctt
3841 gaaagtgcga agaatggtga caccgttgac aggtacacct ttatgggaca ttcccccaga
3901 aaaataatca agactgggcc tttagagggt gctgaagttg accccttggt gcttctggaa
3961 aaagaactga agggcaccag acaagcgcaa cttcctggta ttcctcgtct aagtggtggt
4021 gccataggat acatctcgta cgattgtatt aagtactttg aaccaaaaac tgaaagaaaa
4081 ctgaaagatg ttttgcaact tccggaagca gctttgatgt tgttcgacac gatcgtggct
4141 tttgacaatg tttatcaaag attccaggta attggaaacg tttctctatc cgttgatgac
4201 tcggacgaag ctattcttga gaaatattat aagacaagag aagaagtgga aaagatcagt
4261 aaagtggtat ttgacaataa aactgttccc tactatgaac agaaagatat tattcaaggc
4321 caaacgttca cctctaatat tggtcaggaa gggtatgaaa accatgttcg caagctgaaa
4381 gaacatattc tgaaaggaga catcttccaa gctgttccct ctcaaagggt agccaggccg
4441 acctcattgc accctttcaa catctatcgt catttgagaa ctgtcaatcc ttctccatac
4501 atgttctata ttgactatct agacttccaa gttgttggtg cttcacctga attactagtt
4561 aaatccgaca acaacaacaa aatcatcaca catcctattg ctggaactct tcccagaggt
4621 aaaactatcg aagaggacga caattatgct aagcaattga agtcgtcttt gaaagacagg
4681 gccgagcacg tcatgctggt agatttggcc agaaatgata ttaaccgtgt gtgtgagccc
4741 accagtacca cggttgatcg tttattgact gtggagagat tttctcatgt gatgcatctt
4801 gtgtcagaag tcagtggaac attgagacca aacaagactc gcttcgatgc tttcagatcc
4861 attttcccag caggaaccgt ctccggtgct ccgaaggtaa gagcaatgca actcatagga
4921 gaattggaag gagaaaagag aggtgtttat gcgggggccg taggacactg gtcgtacgat
4981 ggaaaatcga tggacacatg tattgcctta agaacaatgg tcgtcaagga cggtgtcgct
5041 taccttcaag ccggaggtgg aattgtctac gattctgacc cctatgacga gtacatcgaa
5101 accatgaaca aaatgagatc caacaataac accatcttgg aggctgagaa aatctggacc
5161 gataggttgg ccagagacga gaatcaaagt gaatccgaag aaaacgatca atgaacggag
5221 gacgtaagta ggaatttatg gtttggccat aatggcctag cttggcgtaa tcatggtcat
5281 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
5341 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
5401 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
5461 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
5521 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
5581 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
5641 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
5701 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
5761 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
5821 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
5881 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
5941 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
6001 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
6061 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
6121 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
6181 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
6241 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
6301 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
6361 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
6421 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
6481 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
6541 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
6601 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
6661 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
6721 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
6781 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
6841 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
6901 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
6961 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
7021 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
7081 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
7141 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
7201 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
7261 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
7321 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
7381 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
7441 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
7501 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
7561 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
7621 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
7681 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt
7741 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
7801 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
7861 cccagtcacg acgttgtaaa acgacggcca gtgaattg
//