| 出品公司: | Invitrogen |
|---|---|
| 载体名称: | pPIC9, pPIC 9 |
| 质粒类型: | 毕赤酵母蛋白表达载体 |
| 表达水平: | 高拷贝 |
| 启动子: | AOX1 |
| 克隆方法: | 多克隆位点，限制性内切酶 |
| 载体大小: | 8023 bp |
| 5' 测序引物及序列: | 5´ AOX1:5´-GACTGGTTCCAATTGACAAGC-3´； alpha-Factor-F: 5´-TACTATTGCCAGCATTGCTGC-3´ |
| 3' 测序引物及序列: | 3´ AOX1:5´-GCAAATGGCATTCTGACATCC-3´ |
| 载体标签: | / |
| 载体抗性: | Ampicillin |
| 筛选标记: | HIS4 |
| 备注: |
利用alpha分泌因子，分泌表达蛋白。
|
| 产品目录号: | K1710-01 |
| 稳定性: | 稳定 Stable |
| 组成型: | 组成型 Constitutive |
| 病毒/非病毒: | 非病毒 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
pPIC9载体的详细信息如下：
• pPIC9载体大小8,023 bp，是融合表达载体。
• 载体构建过程中，可供使用的单一限制性内切酶位点为Xho I, SnaB I, EcoR I, Avr II, Not I.
• 利用alpha因子分泌信号肽，分泌表达蛋白基因。
• 在载体构建过程中，你的基因必须保证与信号肽的起始密码子的读码框一致。
• 毕赤酵母中利用HIS4进行筛选。
• 为了将基因插入GS115或KM71的AOX区, 使用SacI限制性内切酶线性化质粒(GS115中产生His+Mut+基因型，KM71中产生His+ MutS基因型)
•为了将基因插入GS115或KM71的His4区, 使用Sal I或者Stu I限制性内切酶线性化质粒(GS115中产生His+ Mut+基因型，KM71中产生His+ MutS基因型)
•将基因插入GS115的AOX1区域，需要使用Bgl II线性化质粒 (产生His+MutS基因型)。
载体序列
LOCUS pPIC9 8023 bp ds-DNA circular SYN
DEFINITION Pichia pastoris HIS4 vector for methanol-inducible expression of a
secreted protein.
ACCESSION .
VERSION .
KEYWORDS pPIC9
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT VNTAUTHORNAME|biofeng.com|
FEATURES Location/Qualifiers
source 1..8023
/organism="synthetic DNA construct"
/lab_host="Pichia pastoris"
/mol_type="other DNA"
promoter 2..937
/gene="Pichia pastoris AOX1"
/note="AOX1 promoter"
/note="inducible promoter, regulated by methanol"
/note="color: #ffffff; direction: RIGHT"
CDS 949..1215
/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."
/note="This feature has 3 segments:
???1:??949?..?1005?/?#993366?/?presequence
???2:?1006?..?1203?/?#993366?/?pro?region
???3:?1204?..?1215?/?#993366
Cleavage sites after bases 1005, 1203"
/translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA"
misc_feature 1192..1241
/note="MCS"
/note="multiple cloning site"
/note="color: #99ccff"
terminator 1321..1567
/gene="Pichia pastoris AOX1"
/note="AOX1 terminator"
/note="transcription terminator for AOX1"
/note="color: #ffffff"
CDS complement(1980..4514)
/codon_start=1
/gene="Pichia pastoris HIS4"
/locus_tag="
"
/product="multifunctional enzyme, required for histidine
biosynthesis"
/note="PpHIS4"
/note="auxotrophic marker for Pichia pastoris"
/note="color: #ff7f50"
/protein_id="
"
/translation="MTFPLLPAYASVAEFDNSLSLVGKAVFPYAADQLHNLIKFTQSTE
LQVNVQVESSVTEDQFEELIDNLLKLYNNGINEVILDLDLAERVVQRMIPGARVIYRTL
VDKVASLPANASIAVPFSSPLGDLKSFTNGGSRTVYAFSETAKLVDVTSTVASGIIPII
DARQLTTEYELSEDVKKFPVSEILLASLTTDRPDGLFTTLVADSSNYSLGLVYSSKKSI
PEAIRTQTGVYQSRRHGLWYKGATSGATQKLLGIELDCDGDCLKFVVEQTGVGFCHLER
TSCFGQSKGLRAMEATLWDRKSNAPEGSYTKRLFDDEVLLNAKIREEADELAEAKSKED
IAWECADLFYFALVRCAKYGVTLDEVERNLDMKSLKVTRRKGDAKPGYTKEQPKEESKP
KEVPSEGRIELCKIDVSKASSQEIEDALRRPIQKTEQIMELVKPIVDNVRQNGDKALLE
LTAKFDGVALKTPVLEAPFPEELMQLPDNVKRAIDLSIDNVRKFHEAQLTETLQVETCP
GVVCSRFARPIEKVGLYIPGGTAILPSTSLMLGVPAKVAGCKEIVFASPPKKDGTLTPE
VIYVAHKVGAKCIVLAGGAQAVAAMAYGTETVPKCDKIFGPGNQFVTAAKMMVQNDTSA
LCSIDMPAGPSEVLVIADKYADPDFVASDLLSQAEHGIDSQVILLAVDMTDKELARIED
AVHNQAVQLPRVEIVRKCIAHSTTLSVATYEQALEMSNQYAPEHLILQIENASSYVDQV
QHAGSVFVGAYSPESCGDYSSGTNHTLPTYGYARQYSGVNTATFQKFITSQDVTPEGLK
HIGQAVMDLAAVEGLDAHRNAVKVRMEKLGLI"
misc_feature 4869..5625
/note="AOX1 3' fragment"
/note="region downstream of Pichia pastoris AOX1 gene"
/note="color: #ffffff"
rep_origin complement(6094..6682)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
/note="color: #ffff00"
CDS complement(6853..7713)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/note="This feature has 2 segments:
???1:?6853?..?7644?/?#ccffcc
???2:?7645?..?7713?/?#ccffcc?/?signal?sequence
Cleavage site after base 7644"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(7714..7818)
/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 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 cgaattaatt cgccttagac
1261 atgactgttc ctcagttcaa gttgggcact tacgagaaga ccggtcttgc tagattctaa
1321 tcaagaggat gtcagaatgc catttgcctg agagatgcag gcttcatttt tgatactttt
1381 ttatttgtaa cctatatagt ataggatttt ttttgtcatt ttgtttcttc tcgtacgagc
1441 ttgctcctga tcagcctatc tcgcagctga tgaatatctt gtggtagggg tttgggaaaa
1501 tcattcgagt ttgatgtttt tcttggtatt tcccactcct cttcagagta cagaagatta
1561 agtgagaagt tcgtttgtgc aagcttatcg ataagcttta atgcggtagt ttatcacagt
1621 taaattgcta acgcagtcag gcaccgtgta tgaaatctaa caatgcgctc atcgtcatcc
1681 tcggcaccgt caccctggat gctgtaggca taggcttggt tatgccggta ctgccgggcc
1741 tcttgcggga tatcgtccat tccgacagca tcgccagtca ctatggcgtg ctgctagcgc
1801 tatatgcgtt gatgcaattt ctatgcgcac ccgttctcgg agcactgtcc gaccgctttg
1861 gccgccgccc agtcctgctc gcttcgctac ttggagccac tatcgactac gcgatcatgg
1921 cgaccacacc cgtcctgtgg atctatcgaa tctaaatgta agttaaaatc tctaaataat
1981 taaataagtc ccagtttctc catacgaacc ttaacagcat tgcggtgagc atctagacct
2041 tcaacagcag ccagatccat cactgcttgg ccaatatgtt tcagtccctc aggagttacg
2101 tcttgtgaag tgatgaactt ctggaaggtt gcagtgttaa ctccgctgta ttgacgggca
2161 tatccgtacg ttggcaaagt gtggttggta ccggaggagt aatctccaca actctctgga
2221 gagtaggcac caacaaacac agatccagcg tgttgtactt gatcaacata agaagaagca
2281 ttctcgattt gcaggatcaa gtgttcagga gcgtactgat tggacatttc caaagcctgc
2341 tcgtaggttg caaccgatag ggttgtagag tgtgcaatac acttgcgtac aatttcaacc
2401 cttggcaact gcacagcttg gttgtgaaca gcatcttcaa ttctggcaag ctccttgtct
2461 gtcatatcga cagccaacag aatcacctgg gaatcaatac catgttcagc ttgagacaga
2521 aggtctgagg caacgaaatc tggatcagcg tatttatcag caataactag aacttcagaa
2581 ggcccagcag gcatgtcaat actacacagg gctgatgtgt cattttgaac catcatcttg
2641 gcagcagtaa cgaactggtt tcctggacca aatattttgt cacacttagg aacagtttct
2701 gttccgtaag ccatagcagc tactgcctgg gcgcctcctg ctagcacgat acacttagca
2761 ccaaccttgt gggcaacgta gatgacttct ggggtaaggg taccatcctt cttaggtgga
2821 gatgcaaaaa caatttcttt gcaaccagca actttggcag gaacacccag catcagggaa
2881 gtggaaggca gaattgcggt tccaccagga atatagaggc caactttctc aataggtctt
2941 gcaaaacgag agcagactac accagggcaa gtctcaactt gcaacgtctc cgttagttga
3001 gcttcatgga atttcctgac gttatctata gagagatcaa tggctctctt aacgttatct
3061 ggcaattgca taagttcctc tgggaaagga gcttctaaca caggtgtctt caaagcgact
3121 ccatcaaact tggcagttag ttctaaaagg gctttgtcac cattttgacg aacattgtcg
3181 acaattggtt tgactaattc cataatctgt tccgttttct ggataggacg acgaagggca
3241 tcttcaattt cttgtgagga ggccttagaa acgtcaattt tgcacaattc aatacgacct
3301 tcagaaggga cttctttagg tttggattct tctttaggtt gttccttggt gtatcctggc
3361 ttggcatctc ctttccttct agtgaccttt agggacttca tatccaggtt tctctccacc
3421 tcgtccaacg tcacaccgta cttggcacat ctaactaatg caaaataaaa taagtcagca
3481 cattcccagg ctatatcttc cttggattta gcttctgcaa gttcatcagc ttcctcccta
3541 attttagcgt tcaacaaaac ttcgtcgtca aataaccgtt tggtataaga accttctgga
3601 gcattgctct tacgatccca caaggtggct tccatggctc taagaccctt tgattggcca
3661 aaacaggaag tgcgttccaa gtgacagaaa ccaacacctg tttgttcaac cacaaatttc
3721 aagcagtctc catcacaatc caattcgata cccagcaact tttgagttgc tccagatgta
3781 gcacctttat accacaaacc gtgacgacga gattggtaga ctccagtttg tgtccttata
3841 gcctccggaa tagacttttt ggacgagtac accaggccca acgagtaatt agaagagtca
3901 gccaccaaag tagtgaatag accatcgggg cggtcagtag tcaaagacgc caacaaaatt
3961 tcactgacag ggaacttttt gacatcttca gaaagttcgt attcagtagt caattgccga
4021 gcatcaataa tggggattat accagaagca acagtggaag tcacatctac caactttgcg
4081 gtctcagaaa aagcataaac agttctacta ccgccattag tgaaactttt caaatcgccc
4141 agtggagaag aaaaaggcac agcgatacta gcattagcgg gcaaggatgc aactttatca
4201 accagggtcc tatagataac cctagcgcct gggatcatcc tttggacaac tctttctgcc
4261 aaatctaggt ccaaaatcac ttcattgata ccattattgt acaacttgag caagttgtcg
4321 atcagctcct caaattggtc ctctgtaacg gatgactcaa cttgcacatt aacttgaagc
4381 tcagtcgatt gagtgaactt gatcaggttg tgcagctggt cagcagcata gggaaacacg
4441 gcttttccta ccaaactcaa ggaattatca aactctgcaa cacttgcgta tgcaggtagc
4501 aagggaaatg tcatacttga agtcggacag tgagtgtagt cttgagaaat tctgaagccg
4561 tatttttatt atcagtgagt cagtcatcag gagatcctct acgccggacg catcgtggcc
4621 ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat caccgatggg
4681 gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg tatggtggca
4741 ggccccgtgg ccgggggact gttgggcgcc atctccttgc atgcaccatt ccttgcggcg
4801 gcggtgctca acggcctcaa cctactactg ggctgcttcc taatgcagga gtcgcataag
4861 ggagagcgtc gagtatctat gattggaagt atgggaatgg tgatacccgc attcttcagt
4921 gtcttgaggt ctcctatcag attatgccca actaaagcaa ccggaggagg agatttcatg
4981 gtaaatttct ctgacttttg gtcatcagta gactcgaact gtgagactat ctcggttatg
5041 acagcagaaa tgtccttctt ggagacagta aatgaagtcc caccaataaa gaaatccttg
5101 ttatcaggaa caaacttctt gtttcgaact ttttcggtgc cttgaactat aaaatgtaga
5161 gtggatatgt cgggtaggaa tggagcgggc aaatgcttac cttctggacc ttcaagaggt
5221 atgtagggtt tgtagatact gatgccaact tcagtgacaa cgttgctatt tcgttcaaac
5281 cattccgaat ccagagaaat caaagttgtt tgtctactat tgatccaagc cagtgcggtc
5341 ttgaaactga caatagtgtg ctcgtgtttt gaggtcatct ttgtatgaat aaatctagtc
5401 tttgatctaa ataatcttga cgagccaagg cgataaatac ccaaatctaa aactctttta
5461 aaacgttaaa aggacaagta tgtctgcctg tattaaaccc caaatcagct cgtagtctga
5521 tcctcatcaa cttgaggggc actatcttgt tttagagaaa tttgcggaga tgcgatatcg
5581 agaaaaaggt acgctgattt taaacgtgaa atttatctca agatctctgc ctcgcgcgtt
5641 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
5701 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
5761 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta
5821 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
5881 atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct
5941 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
6001 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
6061 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
6121 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
6181 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
6241 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
6301 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
6361 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
6421 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
6481 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
6541 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
6601 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
6661 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
6721 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
6781 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
6841 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
6901 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
6961 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
7021 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
7081 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
7141 tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg
7201 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
7261 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
7321 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
7381 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
7441 gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac
7501 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
7561 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
7621 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
7681 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
7741 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
7801 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
7861 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcaaga
7921 attaattctc atgtttgaca gcttatcatc gataagctga ctcatgttgg tattgtgaaa
7981 tagacgcaga tcgggaacac tgaaaaataa cagttattat tcg
//
DEFINITION Pichia pastoris HIS4 vector for methanol-inducible expression of a
secreted protein.
ACCESSION .
VERSION .
KEYWORDS pPIC9
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT VNTAUTHORNAME|biofeng.com|
FEATURES Location/Qualifiers
source 1..8023
/organism="synthetic DNA construct"
/lab_host="Pichia pastoris"
/mol_type="other DNA"
promoter 2..937
/gene="Pichia pastoris AOX1"
/note="AOX1 promoter"
/note="inducible promoter, regulated by methanol"
/note="color: #ffffff; direction: RIGHT"
CDS 949..1215
/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."
/note="This feature has 3 segments:
???1:??949?..?1005?/?#993366?/?presequence
???2:?1006?..?1203?/?#993366?/?pro?region
???3:?1204?..?1215?/?#993366
Cleavage sites after bases 1005, 1203"
/translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA"
misc_feature 1192..1241
/note="MCS"
/note="multiple cloning site"
/note="color: #99ccff"
terminator 1321..1567
/gene="Pichia pastoris AOX1"
/note="AOX1 terminator"
/note="transcription terminator for AOX1"
/note="color: #ffffff"
CDS complement(1980..4514)
/codon_start=1
/gene="Pichia pastoris HIS4"
/locus_tag="
"
/product="multifunctional enzyme, required for histidine
biosynthesis"
/note="PpHIS4"
/note="auxotrophic marker for Pichia pastoris"
/note="color: #ff7f50"
/protein_id="
"
/translation="MTFPLLPAYASVAEFDNSLSLVGKAVFPYAADQLHNLIKFTQSTE
LQVNVQVESSVTEDQFEELIDNLLKLYNNGINEVILDLDLAERVVQRMIPGARVIYRTL
VDKVASLPANASIAVPFSSPLGDLKSFTNGGSRTVYAFSETAKLVDVTSTVASGIIPII
DARQLTTEYELSEDVKKFPVSEILLASLTTDRPDGLFTTLVADSSNYSLGLVYSSKKSI
PEAIRTQTGVYQSRRHGLWYKGATSGATQKLLGIELDCDGDCLKFVVEQTGVGFCHLER
TSCFGQSKGLRAMEATLWDRKSNAPEGSYTKRLFDDEVLLNAKIREEADELAEAKSKED
IAWECADLFYFALVRCAKYGVTLDEVERNLDMKSLKVTRRKGDAKPGYTKEQPKEESKP
KEVPSEGRIELCKIDVSKASSQEIEDALRRPIQKTEQIMELVKPIVDNVRQNGDKALLE
LTAKFDGVALKTPVLEAPFPEELMQLPDNVKRAIDLSIDNVRKFHEAQLTETLQVETCP
GVVCSRFARPIEKVGLYIPGGTAILPSTSLMLGVPAKVAGCKEIVFASPPKKDGTLTPE
VIYVAHKVGAKCIVLAGGAQAVAAMAYGTETVPKCDKIFGPGNQFVTAAKMMVQNDTSA
LCSIDMPAGPSEVLVIADKYADPDFVASDLLSQAEHGIDSQVILLAVDMTDKELARIED
AVHNQAVQLPRVEIVRKCIAHSTTLSVATYEQALEMSNQYAPEHLILQIENASSYVDQV
QHAGSVFVGAYSPESCGDYSSGTNHTLPTYGYARQYSGVNTATFQKFITSQDVTPEGLK
HIGQAVMDLAAVEGLDAHRNAVKVRMEKLGLI"
misc_feature 4869..5625
/note="AOX1 3' fragment"
/note="region downstream of Pichia pastoris AOX1 gene"
/note="color: #ffffff"
rep_origin complement(6094..6682)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
/note="color: #ffff00"
CDS complement(6853..7713)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/note="This feature has 2 segments:
???1:?6853?..?7644?/?#ccffcc
???2:?7645?..?7713?/?#ccffcc?/?signal?sequence
Cleavage site after base 7644"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(7714..7818)
/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 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 cgaattaatt cgccttagac
1261 atgactgttc ctcagttcaa gttgggcact tacgagaaga ccggtcttgc tagattctaa
1321 tcaagaggat gtcagaatgc catttgcctg agagatgcag gcttcatttt tgatactttt
1381 ttatttgtaa cctatatagt ataggatttt ttttgtcatt ttgtttcttc tcgtacgagc
1441 ttgctcctga tcagcctatc tcgcagctga tgaatatctt gtggtagggg tttgggaaaa
1501 tcattcgagt ttgatgtttt tcttggtatt tcccactcct cttcagagta cagaagatta
1561 agtgagaagt tcgtttgtgc aagcttatcg ataagcttta atgcggtagt ttatcacagt
1621 taaattgcta acgcagtcag gcaccgtgta tgaaatctaa caatgcgctc atcgtcatcc
1681 tcggcaccgt caccctggat gctgtaggca taggcttggt tatgccggta ctgccgggcc
1741 tcttgcggga tatcgtccat tccgacagca tcgccagtca ctatggcgtg ctgctagcgc
1801 tatatgcgtt gatgcaattt ctatgcgcac ccgttctcgg agcactgtcc gaccgctttg
1861 gccgccgccc agtcctgctc gcttcgctac ttggagccac tatcgactac gcgatcatgg
1921 cgaccacacc cgtcctgtgg atctatcgaa tctaaatgta agttaaaatc tctaaataat
1981 taaataagtc ccagtttctc catacgaacc ttaacagcat tgcggtgagc atctagacct
2041 tcaacagcag ccagatccat cactgcttgg ccaatatgtt tcagtccctc aggagttacg
2101 tcttgtgaag tgatgaactt ctggaaggtt gcagtgttaa ctccgctgta ttgacgggca
2161 tatccgtacg ttggcaaagt gtggttggta ccggaggagt aatctccaca actctctgga
2221 gagtaggcac caacaaacac agatccagcg tgttgtactt gatcaacata agaagaagca
2281 ttctcgattt gcaggatcaa gtgttcagga gcgtactgat tggacatttc caaagcctgc
2341 tcgtaggttg caaccgatag ggttgtagag tgtgcaatac acttgcgtac aatttcaacc
2401 cttggcaact gcacagcttg gttgtgaaca gcatcttcaa ttctggcaag ctccttgtct
2461 gtcatatcga cagccaacag aatcacctgg gaatcaatac catgttcagc ttgagacaga
2521 aggtctgagg caacgaaatc tggatcagcg tatttatcag caataactag aacttcagaa
2581 ggcccagcag gcatgtcaat actacacagg gctgatgtgt cattttgaac catcatcttg
2641 gcagcagtaa cgaactggtt tcctggacca aatattttgt cacacttagg aacagtttct
2701 gttccgtaag ccatagcagc tactgcctgg gcgcctcctg ctagcacgat acacttagca
2761 ccaaccttgt gggcaacgta gatgacttct ggggtaaggg taccatcctt cttaggtgga
2821 gatgcaaaaa caatttcttt gcaaccagca actttggcag gaacacccag catcagggaa
2881 gtggaaggca gaattgcggt tccaccagga atatagaggc caactttctc aataggtctt
2941 gcaaaacgag agcagactac accagggcaa gtctcaactt gcaacgtctc cgttagttga
3001 gcttcatgga atttcctgac gttatctata gagagatcaa tggctctctt aacgttatct
3061 ggcaattgca taagttcctc tgggaaagga gcttctaaca caggtgtctt caaagcgact
3121 ccatcaaact tggcagttag ttctaaaagg gctttgtcac cattttgacg aacattgtcg
3181 acaattggtt tgactaattc cataatctgt tccgttttct ggataggacg acgaagggca
3241 tcttcaattt cttgtgagga ggccttagaa acgtcaattt tgcacaattc aatacgacct
3301 tcagaaggga cttctttagg tttggattct tctttaggtt gttccttggt gtatcctggc
3361 ttggcatctc ctttccttct agtgaccttt agggacttca tatccaggtt tctctccacc
3421 tcgtccaacg tcacaccgta cttggcacat ctaactaatg caaaataaaa taagtcagca
3481 cattcccagg ctatatcttc cttggattta gcttctgcaa gttcatcagc ttcctcccta
3541 attttagcgt tcaacaaaac ttcgtcgtca aataaccgtt tggtataaga accttctgga
3601 gcattgctct tacgatccca caaggtggct tccatggctc taagaccctt tgattggcca
3661 aaacaggaag tgcgttccaa gtgacagaaa ccaacacctg tttgttcaac cacaaatttc
3721 aagcagtctc catcacaatc caattcgata cccagcaact tttgagttgc tccagatgta
3781 gcacctttat accacaaacc gtgacgacga gattggtaga ctccagtttg tgtccttata
3841 gcctccggaa tagacttttt ggacgagtac accaggccca acgagtaatt agaagagtca
3901 gccaccaaag tagtgaatag accatcgggg cggtcagtag tcaaagacgc caacaaaatt
3961 tcactgacag ggaacttttt gacatcttca gaaagttcgt attcagtagt caattgccga
4021 gcatcaataa tggggattat accagaagca acagtggaag tcacatctac caactttgcg
4081 gtctcagaaa aagcataaac agttctacta ccgccattag tgaaactttt caaatcgccc
4141 agtggagaag aaaaaggcac agcgatacta gcattagcgg gcaaggatgc aactttatca
4201 accagggtcc tatagataac cctagcgcct gggatcatcc tttggacaac tctttctgcc
4261 aaatctaggt ccaaaatcac ttcattgata ccattattgt acaacttgag caagttgtcg
4321 atcagctcct caaattggtc ctctgtaacg gatgactcaa cttgcacatt aacttgaagc
4381 tcagtcgatt gagtgaactt gatcaggttg tgcagctggt cagcagcata gggaaacacg
4441 gcttttccta ccaaactcaa ggaattatca aactctgcaa cacttgcgta tgcaggtagc
4501 aagggaaatg tcatacttga agtcggacag tgagtgtagt cttgagaaat tctgaagccg
4561 tatttttatt atcagtgagt cagtcatcag gagatcctct acgccggacg catcgtggcc
4621 ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat caccgatggg
4681 gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg tatggtggca
4741 ggccccgtgg ccgggggact gttgggcgcc atctccttgc atgcaccatt ccttgcggcg
4801 gcggtgctca acggcctcaa cctactactg ggctgcttcc taatgcagga gtcgcataag
4861 ggagagcgtc gagtatctat gattggaagt atgggaatgg tgatacccgc attcttcagt
4921 gtcttgaggt ctcctatcag attatgccca actaaagcaa ccggaggagg agatttcatg
4981 gtaaatttct ctgacttttg gtcatcagta gactcgaact gtgagactat ctcggttatg
5041 acagcagaaa tgtccttctt ggagacagta aatgaagtcc caccaataaa gaaatccttg
5101 ttatcaggaa caaacttctt gtttcgaact ttttcggtgc cttgaactat aaaatgtaga
5161 gtggatatgt cgggtaggaa tggagcgggc aaatgcttac cttctggacc ttcaagaggt
5221 atgtagggtt tgtagatact gatgccaact tcagtgacaa cgttgctatt tcgttcaaac
5281 cattccgaat ccagagaaat caaagttgtt tgtctactat tgatccaagc cagtgcggtc
5341 ttgaaactga caatagtgtg ctcgtgtttt gaggtcatct ttgtatgaat aaatctagtc
5401 tttgatctaa ataatcttga cgagccaagg cgataaatac ccaaatctaa aactctttta
5461 aaacgttaaa aggacaagta tgtctgcctg tattaaaccc caaatcagct cgtagtctga
5521 tcctcatcaa cttgaggggc actatcttgt tttagagaaa tttgcggaga tgcgatatcg
5581 agaaaaaggt acgctgattt taaacgtgaa atttatctca agatctctgc ctcgcgcgtt
5641 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
5701 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
5761 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta
5821 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
5881 atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct
5941 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
6001 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
6061 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
6121 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
6181 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
6241 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
6301 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
6361 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
6421 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
6481 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
6541 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
6601 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
6661 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
6721 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
6781 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
6841 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
6901 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
6961 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
7021 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
7081 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
7141 tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg
7201 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
7261 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
7321 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
7381 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
7441 gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac
7501 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
7561 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
7621 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
7681 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
7741 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
7801 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
7861 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcaaga
7921 attaattctc atgtttgaca gcttatcatc gataagctga ctcatgttgg tattgtgaaa
7981 tagacgcaga tcgggaacac tgaaaaataa cagttattat tcg
//
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