| 质粒名称: | pGKB |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 85077 |
| 存储实验室: | Sheng Yang |
|
载体骨架基本信息 |
|
| 载体名称: | pGAPZB |
| 载体出品公司: | Invitrogen |
| 空载体大小: | 3000bp |
| 完整质粒大小: | 4989 bp |
| 载体类型: | Yeast Expression |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | 卡那霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 克隆方法: | 限制性内切酶 |
| 5’测序引物 | GAP |
| 3’测序引物 | 3 AOX1 |
|
文献引用方法 |
|
| 材料和方法部分: | pGKB was a gift from Sheng Yang (Addgene plasmid # 85077 ; http://n2t.net/addgene:85077 ; RRID:Addgene_85077) |
| 参考文献部分: | Hygromycin-resistance vectors for gene expression in Pichia pastoris. Yang J, Nie L, Chen B, Liu Y, Kong Y, Wang H, Diao L. Yeast. 2014 Apr;31(4):115-25. PubMed 24822243 |
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
载体序列
LOCUS pGKB 3506 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION expression in Pichia pastoris.
ACCESSION .
VERSION .
KEYWORDS pGKB
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3506)
AUTHORS Yang J, Nie L, Chen B, Liu Y, Kong Y, Wang H, Diao L
TITLE Hygromycin-resistance vectors for gene expression in Pichia
pastoris.
JOURNAL Yeast. 2014 Apr;31(4):115-25.
PUBMED 24822243
REFERENCE 2 (bases 1 to 3506)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..3506
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 58..534
/label=GAP promoter
/note="promoter for Pichia pastoris
glyceraldehyde-3-phosphate dehydrogenase"
CDS 613..642
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 658..675
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
terminator 755..1001
/gene="Pichia pastoris AOX1"
/label=AOX1 terminator
/note="transcription terminator for AOX1"
promoter 1022..1429
/gene="S. cerevisiae TEF1"
/label=TEF1 promoter
/note="promoter for EF-1-alpha"
promoter 1436..1483
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 1502..2311
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind complement(1569..1588)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
terminator 2317..2514
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
terminator 2563..2810
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
primer_bind complement(2579..2597)
/label=CYC1
/note="CYC1 transcription termination signal, reverse
primer"
rep_origin complement(2885..3473)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2965..2984)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 gctcagtgga acgaaaactc acgttaaggg attttggtca tgcatgagat cagatctttt
61 ttgtagaaat gtcttggtgt cctcgtccaa tcaggtagcc atctctgaaa tatctggctc
121 cgttgcaact ccgaacgacc tgctggcaac gtaaaattct ccggggtaaa acttaaatgt
181 ggagtaatgg aaccagaaac gtctcttccc ttctctctcc ttccaccgcc cgttaccgtc
241 cctaggaaat tttactctgc tggagagctt cttctacggc ccccttgcag caatgctctt
301 cccagcatta cgttgcgggt aaaacggagg tcgtgtaccc gacctagcag cccagggatg
361 gaaaagtccc ggccgtcgct ggcaataata gcgggcggac gcatgtcatg agattattgg
421 aaaccaccag aatcgaatat aaaaggcgaa cacctttccc aattttggtt tctcctgacc
481 caaagacttt aaatttaatt tatttgtccc tatttcaatc aattgaacaa ctatttcgaa
541 acgaggaatt cacgtggccc agccggccgt ctcggatcgg tacctcgagc cgcggcggcc
601 gccagctttc tagaacaaaa actcatctca gaagaggatc tgaatagcgc cgtcgaccat
661 catcatcatc atcattgagt ttgtagcctt agacatgact gttcctcagt tcaagttggg
721 cacttacgag aagaccggtc ttgctagatt ctaatcaaga ggatgtcaga atgccatttg
781 cctgagagat gcaggcttca tttttgatac ttttttattt gtaacctata tagtatagga
841 ttttttttgt cattttgttt cttctcgtac gagcttgctc ctgatcagcc tatctcgcag
901 ctgatgaata tcttgtggta ggggtttggg aaaatcattc gagtttgatg tttttcttgg
961 tatttcccac tcctcttcag agtacagaag attaagtgag accttcgttt gtgcggatcc
1021 cccacacacc atagcttcaa aatgtttcta ctcctttttt actcttccag attttctcgg
1081 actccgcgca tcgccgtacc acttcaaaac acccaagcac agcatactaa atttcccctc
1141 tttcttcctc tagggtgtcg ttaattaccc gtactaaagg tttggaaaag aaaaaagaga
1201 ccgcctcgtt tctttttctt cgtcgaaaaa ggcaataaaa atttttatca cgtttctttt
1261 tcttgaaaat tttttttttt gatttttttc tctttcgatg acctcccatt gatatttaag
1321 ttaataaacg gtcttcaatt tctcaagttt cagtttcatt tttcttgttc tattacaact
1381 ttttttactt cttgctcatt agaaagaaag catagcaatc taatctaagg gcggtgttga
1441 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
1501 catgggtaag gaaaagactc acgtttcgag gccgcgatta aattccaaca tggatgctga
1561 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
1621 attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
1681 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
1741 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
1801 cggcaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
1861 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
1921 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
1981 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
2041 gcataagctt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
2101 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
2161 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
2221 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
2281 gtttcatttg atgctcgatg agtttttcta atcagtactg acaataaaaa gattcttgtt
2341 ttcaagaact tgtcatttgt atagtttttt tatattgtag ttgttctatt ttaatcaaat
2401 gttagcgtga tttatatttt ttttcgcctc gacatcatct gcccagatgc gaagttaagt
2461 gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat actgctgtcg
2521 attcgatact aacgccgcca tccagtgtcg aaaacgatat catcatgtaa ttagttatgt
2581 cacgcttaca ttcacgccct ccccccacat ccgctctaac cgaaaaggaa ggagttagac
2641 aacctgaagt ctaggtccct atttattttt ttatagttat gttagtatta agaacgttat
2701 ttatatttca aatttttctt ttttttctgt acagacgcgt gtacgcatgt aacattatac
2761 tgaaaacctt gcttgagaag gttttgggac gctcgaaggc tttaatttgc aagctggaga
2821 ccaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
2881 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
2941 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
3001 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
3061 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
3121 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
3181 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
3241 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
3301 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
3361 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
3421 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
3481 ctttgatctt ttctacgggg tctgac
//
