| 出品公司: | ZYbscience |
|---|---|
| 载体名称: | pESC-URA, pESC URA |
| Gene accession NO. : | AF063585 |
| 质粒类型: | 酿酒酵母蛋白表达载体 |
| 表达水平: | 高拷贝 |
| 诱导方法: | 半乳糖 |
| 启动子: | GAL1和GAL10 |
| 克隆方法: | 多克隆位点,限制性内切酶 |
| 载体大小: | 6631 bp |
| 5' 测序引物及序列: | GAL1-F: ATTTTCGGTTTGTATTACTTC; GAL10-F: GGTGGTAATGCCATGTAATATG |
| 3' 测序引物及序列: | GALI-R: GTTCTTAATACTAACATAACT GAL10-R: GGCAAGGTAGACAAGCCGACAAC |
| 载体标签: | C-Flag, C-Myc |
| 载体抗性: | 氨苄 |
| 筛选标记: | URA3 |
| 备注: | 利用半乳糖诱导,可以同时使两个基因在酿酒酵母中表达。 |
| 产品目录号: | ZY217454 |
| 稳定性: | 稳定 Stable |
| 组成型/诱导型: | 诱导型 |
| 病毒/非病毒: | 非病毒 |
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- 详细信息
- 文献和实验
- 技术资料
- 保存条件:
-20℃低温保存
- 保质期:
三年
- 英文名:
pESC-URA
- 库存:
20
- 供应商:
泽叶生物
载体基本信息
载体质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pESC-URA 6631 bp DNA circular SYN 24-SEP-2005
DEFINITION Cloning vector pESC-URA, complete sequence.
ACCESSION AF063585
VERSION AF063585.2 GI:6446607
SOURCE Cloning vector pESC-URA.
ORGANISM Cloning vector pESC-URA
artificial sequences; vectors.
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT VNTAUTHORNAME|biofeng.com|
FEATURES Location/Qualifiers
source 1..6631
/organism="Cloning vector pESC-URA"
/mol_type="genomic DNA"
/specific_host="Escherichia coli K12"
/db_xref="taxon:76664"
/vntifkey="98"
CDS complement(4232..5089)
/vntifkey="4"
/label=AmpR
rep_origin 5223..6378
/vntifkey="33"
/label=2\micron\ori
CDS 417..1217
/vntifkey="4"
/label=URA3
/note="yeast URA3 ORF"
terminator complement(1885..2049)
/vntifkey="43"
/label=ADH1\terminator
terminator 3038..3227
/vntifkey="43"
/label=CYC1\terminator
rep_origin complement(1483..1789)
/vntifkey="33"
/label=f1\ori
promoter complement(2268..2479)
/vntifkey="29"
/label=GAL10\promoter
promoter 2480..2934
/vntifkey="29"
/label=GAL1\promoter
misc_feature 2976..3011
/vntifkey="21"
/label=c-myc\tag
misc_feature complement(2198..2224)
/vntifkey="21"
/label=FLAG\tag
misc_feature complement(2180..2263)
/vntifkey="21"
/label=MCS1
misc_feature 2936..3033
/vntifkey="21"
/label=MCS2
rep_origin 3414..4081
/vntifkey="33"
/label=pUC\ori
primer 2770..2788
/vntifkey="27"
/label=oligo\SapA
primer 2837..2857
/vntifkey="27"
/label=Sequence\Primer
BASE COUNT 1900 a 1403 c 1474 g 1854 t
ORIGIN
1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
181 accataccac agcttttcaa ttcaattcat catttttttt ttattctttt ttttgatttc
241 ggtttctttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
301 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
361 cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaatcatgt
421 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
481 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
541 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
601 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
661 ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt aatacagtca
721 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
781 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
841 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
901 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
961 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
1021 ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga accgtggatg
1081 atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
1141 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
1201 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac
1261 aaattagagc ttcaatttaa ttatatcagt tattacccta tgcggtgtga aataccgcac
1321 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat
1381 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
1441 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
1501 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
1561 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
1621 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
1681 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
1741 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
1801 gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
1861 ttcgctatta cgccagctga attggagcga cctcatgcta tacctgagaa agcaacctga
1921 cctacaggaa agagttactc aagaataaga attttcgttt taaaacctaa gagtcacttt
1981 aaaatttgta tacacttatt ttttttataa cttatttaat aataaaaatc ataaatcata
2041 agaaattcgc ttatttagaa gtgtcaacaa cgtatctacc aacgatttga cccttttcca
2101 tcttttcgta aatttctggc aaggtagaca agccgacaac cttgattgga gacttgacca
2161 aacctctggc gaagaattgt taattaagag ctcagatctt atcgtcgtca tccttgtaat
2221 ccatcgatac tagtgcggcc gccctttagt gagggttgaa ttcgaatttt caaaaattct
2281 tacttttttt ttggatggac gcaaagaagt ttaataatca tattacatgg cattaccacc
2341 atatacatat ccatatacat atccatatct aatcttactt atatgttgtg gaaatgtaaa
2401 gagccccatt atcttagcct aaaaaaacct tctctttgga actttcagta atacgcttaa
2461 ctgctcattg ctatattgaa gtacggatta gaagccgccg agcgggtgac agccctccga
2521 aggaagactc tcctccgtgc gtcctcgtct tcaccggtcg cgttcctgaa acgcagatgt
2581 gcctcgcgcc gcactgctcc gaacaataaa gattctacaa tactagcttt tatggttatg
2641 aagaggaaaa attggcagta acctggcccc acaaaccttc aaatgaacga atcaaattaa
2701 caaccatagg atgataatgc gattagtttt ttagccttat ttctggggta attaatcagc
2761 gaagcgatga tttttgatct attaacagat atataaatgc aaaaactgca taaccacttt
2821 aactaatact ttcaacattt tcggtttgta ttacttctta ttcaaatgta ataaaagtat
2881 caacaaaaaa ttgttaatat acctctatac tttaacgtca aggagaaaaa accccggatc
2941 cgtaatacga ctcactatag ggcccgggcg tcgacatgga acagaagttg atttccgaag
3001 aagacctcga gtaagcttgg taccgcggct agctaagatc cgctctaacc gaaaaggaag
3061 gagttagaca acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa
3121 gaacgttatt tatatttcaa atttttcttt tttttctgta cagacgcgtg tacgcatgta
3181 acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgaagatc cagctgcatt
3241 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
3301 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
3361 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
3421 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
3481 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
3541 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
3601 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
3661 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
3721 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
3781 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
3841 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
3901 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
3961 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
4021 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
4081 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
4141 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
4201 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
4261 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
4321 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
4381 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
4441 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
4501 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
4561 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
4621 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
4681 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
4741 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
4801 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
4861 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
4921 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
4981 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
5041 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
5101 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
5161 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
5221 aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc gctaattttt
5281 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa agcgctattt
5341 taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa
5401 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc
5461 tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc
5521 gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg ctctataatg
5581 cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg
5641 tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta
5701 gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat
5761 gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag
5821 aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca
5881 ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa
5941 gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg
6001 agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga
6061 tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct cgttacagtc
6121 cggtgcgttt ttggtttttt gaaagtgcgt cttcagagcg cttttggttt tcaaaagcgc
6181 tctgaagttc ctatactttc tagagaatag gaacttcgga ataggaactt caaagcgttt
6241 ccgaaaacga gcgcttccga aaatgcaacg cgagctgcgc acatacagct cactgttcac
6301 gtcgcaccta tatctgcgtg ttgcctgtat atatatatac atgagaagaa cggcatagtg
6361 cgtgtttatg cttaaatgcg tacttatatg cgtctattta tgtaggatga aaggtagtct
6421 agtacctcct gtgatattat cccattccat gcggggtatc gtatgcttcc ttcagcacta
6481 ccctttagct gttctatatg ctgccactcc tcaattggat tagtctcatc cttcaatgct
6541 atcatttcct ttgatattgg atcatactaa gaaaccatta ttatcatgac attaacctat
6601 aaaaataggc gtatcacgag gccctttcgt c
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文献和实验体积无水乙醇;1/10体积3.0M NaAc(pH5.2),于-20℃静置过夜。 13. 离心12500xg×20min,4℃,去除上清,沉淀用70%乙醇洗涤,超净台风干。 l),保存于-20℃。mg/管(用于1次转化反应,约40m14. 干燥的DNA溶于适量体积TE,定量,分装100-200 构建于DNA-BD载体的目的DNA转化酵母感受态细胞 1. 将酵母菌EGY48(p8op-lacZ)接种于5.0ml SD/-Ura液体培养基中,缓振打散菌落
4 蛋白调控的GAL1 序列。这个改造过的LacZ 基因被整合到酵母染色体URA3 位上。而酵母的GAL4 基因和GAL80 基因(Gal80 是Gal4 的负调控因子)被缺失,从而排除了细胞内源调控因子的影响。已经知道在Snf1 和Snf2 之间存在相互作用。结果发现只有同时转化了Snf1 和Snf2 融合表达载体的酵母细胞才有β - 半乳糖苷酶活性,单独转化其中任何一个载体都不能检测出β - 半乳糖苷酶活性。目前发展起来的各种双杂交系统大多是以Fields 等人建立的系统为基础的。这些新系统主要
【资源】提供基因合成,载体构建,定点突变等服务;同时出售各种克隆载体,价格实惠!
) PET23b Amp+ 300元(2-4ug) PESC-URA Amp+ 500元(2-4ug) PET24b Kan+ 300元(2-4ug) PHH21 Amp+ 500元(2-4ug) PET24c
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