| 出品公司: | Invitrogen/ Biofeng |
|---|---|
| 载体名称: | pPIC9K-His, pPIC 9K His |
| 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: | 9.3 kb |
| 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: | N-alpha factor, C-His |
| 载体抗性: | 氨苄和卡那 |
| 筛选标记: | HIS4 |
| 备注: |
利用alpha分泌因子，分泌表达带有His标签的蛋白。生物风销售SC-URA酵母表达筛选培养基，有需要和我们联系！
|
| 产品目录号: | V175-20 |
| 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
pPIC9k-His载体质粒基本信息
pPIC9k-His质粒图谱载体图谱和pPIC9k-His载体序列质粒序列多克隆位点信息
pPIC9k-His质粒载体简介
pPIC9K-His载体中存在卡那抗性基因，允许利用卡那抗性在酵母体内筛选多克隆拷贝，载体其余部分与pPIC9载体完全一样。pPIC9K-His载体能够使用的毕赤酵母宿主菌是KM71和GS115。
pPIC9K-His载体详细信息如下：
• pPIC9K-His载体大小9.3 bp，是融合表达载体, 含有C-His标签，表达出来的蛋白含有C端his标签，可以直接用于镍柱纯化。
• 载体构建过程中，可供使用的单一限制性内切酶位点为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Sac 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)
•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)。
pPIC9k-His质粒序列载体序列
LOCUS pPIC9K-His 9294 bp DNA circular 25-DEC-2016
DEFINITION pPIC9K-His.
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTDATE|-11561372|
COMMENT VNTDBDATE|-11561372|
COMMENT LSOWNER|
COMMENT VNTNAME|pPIC9K-His|
COMMENT VNTAUTHORNAME|Demo User|
COMMENT VNTOAUTHORNAME|UNKNOWN|
FEATURES Location/Qualifiers
misc_feature 1152..1172
/vntifkey="21"
/label=alpha_factor_primer
misc_feature complement(1345..1365)
/vntifkey="21"
/label=3_AOX1_primer
gene complement(1668..1955)
/gene="tet(288-0)"
/vntifkey="60"
CDS complement(1998..4532)
/gene="HIS4"
/vntifkey="4"
/label=HIS4
/note="ORF frame 3"
gene complement(1998..4532)
/gene="HIS4"
/vntifkey="60"
CDS complement(4946..5761)
/gene="KanR2"
/vntifkey="4"
/label=Kana\Resistance
/note="ORF frame 2"
gene complement(4946..5761)
/gene="KanR2"
/vntifkey="60"
misc_feature complement(5926..5945)
/vntifkey="21"
/label=pBRrevBam_primer
CDS complement(5927..6649)
/vntifkey="4"
/label=ORF\frame\2
misc_feature complement(6931..6953)
/vntifkey="21"
/label=pGEX_3_primer
rep_origin complement(7306..7979)
/vntifkey="33"
/label=pBR322_origin
CDS complement(8124..8984)
/gene="Ampicillin"
/vntifkey="4"
/label=Ampicillin
/note="ORF frame 3"
gene complement(8124..8984)
/gene="Ampicillin"
/vntifkey="60"
promoter complement(9026..9054)
/vntifkey="30"
/label=AmpR_promoter
promoter 1..948
/vntifkey="29"
/label=5'\AOX1\Promoter\Fragment
primer 855..875
/vntifkey="27"
/label=5'\AOX1\Primer
sig_peptide 949..1218
/vntifkey="94"
/label=alpha\Factor\Secretion\Signal
misc_feature 1192..1241
/vntifkey="21"
/label=Multiple\Cloing\Site
misc_feature 1271..1604
/vntifkey="21"
/label=3'\AOX1\transcription\termination\(TT)
misc_feature 6140..6897
/vntifkey="21"
/label=3'\AOX1\Fragment
misc_feature 1246..1263
/vntifkey="21"
/label=His\Tag
BASE COUNT 2499 a 2205 c 2078 g 2512 t
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 cgtcgtcaaa taaccgtttg
3601 gtataagaac cttctggagc attgctctta cgatcccaca aggtggcttc catggctcta
3661 agaccctttg attggccaaa acaggaagtg cgttccaagt gacagaaacc aacacctgtt
3721 tgttcaacca caaatttcaa gcagtctcca tcacaatcca attcgatacc cagcaacttt
3781 tgagttgctc cagatgtagc acctttatac cacaaaccgt gacgacgaga ttggtagact
3841 ccagtttgtg tccttatagc ctccggaata gactttttgg acgagtacac caggcccaac
3901 gagtaattag aagagtcagc caccaaagta gtgaatagac catcggggcg gtcagtagtc
3961 aaagacgcca acaaaatttc actgacaggg aactttttga catcttcaga aagttcgtat
4021 tcagtagtca attgccgagc atcaataatg gggattatac cagaagcaac agtggaagtc
4081 acatctacca actttgcggt ctcagaaaaa gcataaacag ttctactacc gccattagtg
4141 aaacttttca aatcgcccag tggagaagaa aaaggcacag cgatactagc attagcgggc
4201 aaggatgcaa ctttatcaac cagggtccta tagataaccc tagcgcctgg gatcatcctt
4261 tggacaactc tttctgccaa atctaggtcc aaaatcactt cattgatacc attattgtac
4321 aacttgagca agttgtcgat cagctcctca aattggtcct ctgtaacgga tgactcaact
4381 tgcacattaa cttgaagctc agtcgattga gtgaacttga tcaggttgtg cagctggtca
4441 gcagcatagg gaaacacggc ttttcctacc aaactcaagg aattatcaaa ctctgcaaca
4501 cttgcgtatg caggtagcaa gggaaatgtc atacttgaag tcggacagtg agtgtagtct
4561 tgagaaattc tgaagccgta tttttattat cagtgagtca gtcatcagga gatcctctac
4621 gccggacgca tcgtggccga cctgcagggg gggggggggc gctgaggtct gcctcgtgaa
4681 gaaggtgttg ctgactcata ccaggcctga atcgccccat catccagcca gaaagtgagg
4741 gagccacggt tgatgagagc tttgttgtag gtggaccagt tggtgatttt gaacttttgc
4801 tttgccacgg aacggtctgc gttgtcggga agatgcgtga tctgatcctt caactcagca
4861 aaagttcgat ttattcaaca aagccgccgt cccgtcaagt cagcgtaatg ctctgccagt
4921 gttacaacca attaaccaat tctgattaga aaaactcatc gagcatcaaa tgaaactgca
4981 atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag
5041 gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc
5101 cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa
5161 gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagc ttatgcattt
5221 ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa
5281 ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa
5341 aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa
5401 caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga
5461 tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa
5521 gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa
5581 cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat
5641 agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag
5701 catccatgtt ggaatttaat cgcggcctcg agcaagacgt ttcccgttga atatggctca
5761 taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat
5821 ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtggctt tccccccccc
5881 ccctgcaggt cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca
5941 tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg
6001 gtatggtggc aggccccgtg gccgggggac tgttgggcgc catctccttg catgcaccat
6061 tccttgcggc ggcggtgctc aacggcctca acctactact gggctgcttc ctaatgcagg
6121 agtcgcataa gggagagcgt cgagtatcta tgattggaag tatgggaatg gtgatacccg
6181 cattcttcag tgtcttgagg tctcctatca gattatgccc aactaaagca accggaggag
6241 gagatttcat ggtaaatttc tctgactttt ggtcatcagt agactcgaac tgtgagacta
6301 tctcggttat gacagcagaa atgtccttct tggagacagt aaatgaagtc ccaccaataa
6361 agaaatcctt gttatcagga acaaacttct tgtttcgaac tttttcggtg ccttgaacta
6421 taaaatgtag agtggatatg tcgggtagga atggagcggg caaatgctta ccttctggac
6481 cttcaagagg tatgtagggt ttgtagatac tgatgccaac ttcagtgaca acgttgctat
6541 ttcgttcaaa ccattccgaa tccagagaaa tcaaagttgt ttgtctacta ttgatccaag
6601 ccagtgcggt cttgaaactg acaatagtgt gctcgtgttt tgaggtcatc tttgtatgaa
6661 taaatctagt ctttgatcta aataatcttg acgagccaag gcgataaata cccaaatcta
6721 aaactctttt aaaacgttaa aaggacaagt atgtctgcct gtattaaacc ccaaatcagc
6781 tcgtagtctg atcctcatca acttgagggg cactatcttg ttttagagaa atttgcggag
6841 atgcgatatc gagaaaaagg tacgctgatt ttaaacgtga aatttatctc aagatctctg
6901 cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
6961 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
7021 tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtatac
7081 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
7141 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc
7201 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
7261 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
7321 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
7381 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
7441 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
7501 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa
7561 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
7621 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
7681 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
7741 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
7801 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
7861 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
7921 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
7981 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
8041 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
8101 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
8161 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
8221 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
8281 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
8341 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
8401 tcgccagtta atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc
8461 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
8521 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
8581 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
8641 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
8701 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca
8761 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
8821 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
8881 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
8941 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
9001 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
9061 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
9121 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
9181 cgtcttcaag aattaattct catgtttgac agcttatcat cgataagctg actcatgttg
9241 gtattgtgaa atagacgcag atcgggaaca ctgaaaaata acagttatta ttcg
//
