| 出品公司: | Invitrogen |
|---|---|
| 載體名稱: | pGAPZalpha C, pGAPZα C, pGAPZalphaC, pGAPZαC |
| 質粒類型: | 畢赤酵母蛋白表達載體 |
| 表達水平: | 高拷貝 |
| 啟動子: | GAP |
| 克隆方法: | 多克隆位點，限制性內切酶 |
| 載體大小: | 3152 bp |
| 5' 測序引物及序列: | alpha-Factor-F: 5´-TACTATTGCCAGCATTGCTGC-3´; pGAP-F:5´-GTCCCTATTTCAATCAATTGAA-3´ |
| 3' 測序引物及序列: | 3´ AOX1:5´-GCAAATGGCATTCTGACATCC-3´ |
| 載體標簽: | C-Myc, C-His |
| 載體抗性: | Zeocin 博來霉素 |
| 篩選標記: | HIS4/ Zeocin |
| 備注: |
插入基因是必需包含起始密碼子ATG。
|
| 產品目錄號: | V200-20 |
| 穩定性: | 穩定 Stable |
| 組成型: | 組成型 Constitutive |
| 病毒/非病毒: | 非病毒 |
買家導航
pGAPZα C載體 pGAPZalpha C質粒基本信息
pGAPZα C質粒圖譜 pGAPZalpha C載體圖譜和pGAPZα C載體序列 pGAPZalpha C質粒序列多克隆位點信息
pGAPZα C質粒 pGAPZalpha C載體簡介
甘油醛-3-磷酸脫氫酶（GAPDH）的酶在許多生物細胞中包括畢赤酵母，穩定高水平表達。 最近，GAPDH蛋白編碼基因啟動子（GAP）被深入研究，并在畢赤酵母中能夠高水表達重組蛋白，這取決于使用的碳源（Waterham等 人，，1997）。GAP啟動子（PGAP）的重組蛋白表達水平略高于AOX1啟動子。
pGAPZ A，B，和C載體（2.9 KB）和pGAPZα，B，和C（3.1 KB）載體使用GAP啟動子在畢赤酵母中穩定表達重組蛋白。重組蛋白表達為為帶有C-末端myc抗原表位和C端His標簽的融合蛋白。此外，pGAPZα 表達的融合蛋白在N-端帶有釀酒酵母（Saccharomyces cerevisiae）的α-因子分泌信號肽。這兩個系列的載體都提供三個不同讀碼框的載體，以方便克隆的帶有C-端標簽和/或N-末端分泌信號肽的載體 中。
這些載體是基于顯性選擇標記，Zeocin（博萊霉素）的篩選標記可以同時使用于畢赤酵母和大腸桿菌。
pGAPZα C質粒序列 pGAPZalpha C載體序列
LOCUS pGAPZα C 3152 bp ds-DNA circular SYN 05-二月-2013
DEFINITION Pichia pastoris vector for constitutive expression of a secreted
protein.
ACCESSION .
VERSION .
KEYWORDS pGAPZ(alpha) C
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3152)
AUTHORS Invitrogen (Life Technologies)
TITLE Direct Submission
JOURNAL Exported 2013年7月30日 from SnapGene Viewer 1.5.3
//www.snapgene.com
COMMENT Linearize within the GAP promoter for integration.
FEATURES Location/Qualifiers
source 1..3152
/organism="synthetic DNA construct"
/lab_host="Pichia pastoris"
/mol_type="other DNA"
promoter 7..483
/note="GAP promoter"
/note="promoter for Pichia pastoris
glyceraldehyde-3-phosphate dehydrogenase"
/note="color: #ffffff; direction: RIGHT"
CDS 493..759
/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:?493?..?549?/?#993366?/?presequence
???2:?550?..?747?/?#993366?/?pro?region
???3:?748?..?759?/?#993366
Cleavage sites after bases 549, 747"
/translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA"
misc_feature 759..833
/note="MCS"
/note="multiple cloning site"
/note="color: #99ccff"
CDS 832..861
/codon_start=1
/product="Myc (human c-Myc oncogene) epitope tag"
/note="Myc"
/note="color: #cc99b2"
/translation="EQKLISEEDL"
CDS 877..894
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/note="color: #cc99b2"
/translation="HHHHHH"
terminator 973..1219
/gene="Pichia pastoris AOX1"
/note="AOX1 terminator"
/note="transcription terminator for AOX1"
/note="color: #ffffff"
promoter 1234..1645
/gene="S. cerevisiae TEF1"
/note="TEF1 promoter"
/note="promoter for?EF-1-alpha"
/note="color: #ffffff; direction: RIGHT"
promoter 1652..1699
/note="EM7 promoter"
/note="synthetic bacterial promoter?"
/note="color: #ffffff; direction: RIGHT"
CDS 1718..2092
/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 2158..2405
/gene="S. cerevisiae CYC1"
/note="CYC1 terminator"
/note="transcription terminator for CYC1"
/note="color: #ffffff"
rep_origin complement(2480..3068)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
/note="color: #ffff00"
ORIGIN
1 agatcttttt tgtagaaatg tcttggtgtc ctcgtccaat caggtagcca tctctgaaat
61 atctggctcc gttgcaactc cgaacgacct gctggcaacg taaaattctc cggggtaaaa
121 cttaaatgtg gagtaatgga accagaaacg tctcttccct tctctctcct tccaccgccc
181 gttaccgtcc ctaggaaatt ttactctgct ggagagcttc ttctacggcc cccttgcagc
241 aatgctcttc ccagcattac gttgcgggta aaacggaggt cgtgtacccg acctagcagc
301 ccagggatgg aaaagtcccg gccgtcgctg gcaataatag cgggcggacg catgtcatga
361 gattattgga aaccaccaga atcgaatata aaaggcgaac acctttccca attttggttt
421 ctcctgaccc aaagacttta aatttaattt atttgtccct atttcaatca attgaacaac
481 tatttcgaaa cgatgagatt tccttcaatt tttactgctg ttttattcgc agcatcctcc
541 gcattagctg ctccagtcaa cactacaaca gaagatgaaa cggcacaaat tccggctgaa
601 gctgtcatcg gttactcaga tttagaaggg gatttcgatg ttgctgtttt gccattttcc
661 aacagcacaa ataacgggtt attgtttata aatactacta ttgccagcat tgctgctaaa
721 gaagaagggg tatctctcga gaagagagag gctgaagcat cgatgaattc acgtggccca
781 gccggccgtc tcggatcggt acctcgagcc gcggcggccg ccagctttct agaacaaaaa
841 ctcatctcag aagaggatct gaatagcgcc gtcgaccatc atcatcatca tcattgagtt
901 ttagccttag acatgactgt tcctcagttc aagttgggca cttacgagaa gaccggtctt
961 gctagattct aatcaagagg atgtcagaat gccatttgcc tgagagatgc aggcttcatt
1021 tttgatactt ttttatttgt aacctatata gtataggatt ttttttgtca ttttgtttct
1081 tctcgtacga gcttgctcct gatcagccta tctcgcagct gatgaatatc ttgtggtagg
1141 ggtttgggaa aatcattcga gtttgatgtt tttcttggta tttcccactc ctcttcagag
1201 tacagaagat taagtgagac cttcgtttgt gcggatcccc cacacaccat agcttcaaaa
1261 tgtttctact ccttttttac tcttccagat tttctcggac tccgcgcatc gccgtaccac
1321 ttcaaaacac ccaagcacag catactaaat tttccctctt tcttcctcta gggtgtcgtt
1381 aattacccgt actaaaggtt tggaaaagaa aaaagagacc gcctcgtttc tttttcttcg
1441 tcgaaaaagg caataaaaat ttttatcacg tttctttttc ttgaaatttt tttttttagt
1501 ttttttctct ttcagtgacc tccattgata tttaagttaa taaacggtct tcaatttctc
1561 aagtttcagt ttcatttttc ttgttctatt acaacttttt ttacttcttg ttcattagaa
1621 agaaagcata gcaatctaat ctaagggcgg tgttgacaat taatcatcgg catagtatat
1681 cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt
1741 tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg
1801 gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct
1861 gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt
1921 gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga
1981 cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct
2041 gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtccg
2101 acggcggccc acgggtccca ggcctcggag atccgtcccc cttttccttt gtcgatatca
2161 tgtaattagt tatgtcacgc ttacattcac gccctccccc cacatccgct ctaaccgaaa
2221 aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata gttatgttag
2281 tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg
2341 catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg aaggctttaa
2401 tttgcaagct ggagaccaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
2461 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
2521 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
2581 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
2641 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt
2701 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
2761 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
2821 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
2881 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
2941 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
3001 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
3061 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
3121 cacgttaagg gattttggtc atgcatgaga tc
//