| 质粒名称: | pGHYB |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 87447 |
| 存储实验室: | Sheng Yang |
|
载体骨架基本信息 |
|
| 载体名称: | pGHYB |
| 载体出品公司: | Sheng Yang Lab |
| 空载体大小: | 3500 bp |
| 完整质粒大小: | 3642bp |
| 载体修饰: | Hygromycin |
| 载体类型: | 酵母表达 |
| 载体抗性: | Hygromycin |
| 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: | pGHYB was a gift from Sheng Yang (Addgene plasmid # 87447 ; http://n2t.net/addgene:87447 ; RRID:Addgene_87447) |
| 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 |
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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 pGHYB 3642 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Pichia pastoris expression plasmid.
ACCESSION .
VERSION .
KEYWORDS pGHYB
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3642)
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 3642)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..3642
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 63..310
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
primer_bind complement(79..97)
/label=CYC1
/note="CYC1 transcription termination signal, reverse
primer"
rep_origin complement(385..973)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(465..484)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 1064..1540
/label=GAP promoter
/note="promoter for Pichia pastoris
glyceraldehyde-3-phosphate dehydrogenase"
CDS 1619..1648
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 1664..1681
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
terminator 1761..2007
/gene="Pichia pastoris AOX1"
/label=AOX1 terminator
/note="transcription terminator for AOX1"
gene 2066..3641
/label=hphMX6
/note="yeast selectable marker conferring hygromycin
resistance"
promoter 2066..2409
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
CDS 2410..3438
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
KE"
terminator 3444..3641
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
ORIGIN
1 tgtcgattcg atactaacgc cgccatccag tgtcgaaaac gagctcgaat tcatcgatga
61 tatcatgtaa ttagttatgt cacgcttaca ttcacgccct ccccccacat ccgctctaac
121 cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt ttatagttat
181 gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt acagacgcgt
241 gtacgcatgt aacattatac tgaaaacctt gcttgagaag gttttgggac gctcgaaggc
301 tttaatttgc aagctggaga ccaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
361 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
421 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
481 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
541 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
601 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
661 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
721 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
781 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
841 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
901 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
961 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
1021 aaactcacgt taagggattt tggtcatgca tgagatcaga tcttttttgt agaaatgtct
1081 tggtgtcctc gtccaatcag gtagccatct ctgaaatatc tggctccgtt gcaactccga
1141 acgacctgct ggcaacgtaa aattctccgg ggtaaaactt aaatgtggag taatggaacc
1201 agaaacgtct cttcccttct ctctccttcc accgcccgtt accgtcccta ggaaatttta
1261 ctctgctgga gagcttcttc tacggccccc ttgcagcaat gctcttccca gcattacgtt
1321 gcgggtaaaa cggaggtcgt gtacccgacc tagcagccca gggatggaaa agtcccggcc
1381 gtcgctggca ataatagcgg gcggacgcat gtcatgagat tattggaaac caccagaatc
1441 gaatataaaa ggcgaacacc tttcccaatt ttggtttctc ctgacccaaa gactttaaat
1501 ttaatttatt tgtccctatt tcaatcaatt gaacaactat ttcgaaacga ggaattcacg
1561 tggcccagcc ggccgtctcg gatcggtacc tcgagccgcg gcggccgcca gctttctaga
1621 acaaaaactc atctcagaag aggatctgaa tagcgccgtc gaccatcatc atcatcatca
1681 ttgagtttgt agccttagac atgactgttc ctcagttcaa gttgggcact tacgagaaga
1741 ccggtcttgc tagattctaa tcaagaggat gtcagaatgc catttgcctg agagatgcag
1801 gcttcatttt tgatactttt ttatttgtaa cctatatagt ataggatttt ttttgtcatt
1861 ttgtttcttc tcgtacgagc ttgctcctga tcagcctatc tcgcagctga tgaatatctt
1921 gtggtagggg tttgggaaaa tcattcgagt ttgatgtttt tcttggtatt tcccactcct
1981 cttcagagta cagaagatta agtgagacct tcgtttgtgc ggatctgttt agcttgcctc
2041 gtccccgccg ggtcacccgg ccagcgacat ggaggcccag aataccctcc ttgacagtct
2101 tgacgtgcgc agctcagggg catgatgtga ctgtcgcccg tacatttagc ccatacatcc
2161 ccatgtataa tcatttgcat ccatacattt tgatggccgc acggcgcgaa gcaaaaatta
2221 cggctcctcg ctgcggacct gcgagcaggg aaacgctccc ctcacagacg cgttgaattg
2281 tccccacgcc gcgcccctgt agagaaatat aaaaggttag gatttgccac tgaggttctt
2341 ctttcatata cttcctttta aaatcttgct aggatacagt tctcacatca catccgaaca
2401 taaacaacca tgggtaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc
2461 gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct
2521 ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt
2581 ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa
2641 gtgcttgaca ttggggaatt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag
2701 ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg
2761 gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt cggcccattc
2821 ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat
2881 ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag
2941 gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac
3001 gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac
3061 tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg
3121 ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt
3181 gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag
3241 agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc
3301 gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc
3361 tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt
3421 ccgagggcaa aggaataatc agtactgaca ataaaaagat tcttgttttc aagaacttgt
3481 catttgtata gtttttttat attgtagttg ttctatttta atcaaatgtt agcgtgattt
3541 atattttttt tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa
3601 tatcatgcgt caatcgtatg tgaatgctgg tcgctatact gc
//
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