- ￥100 - 1000
- Xybio
- 上海
- P1269
- 2025年07月14日
9 年钻石会员
企业认证
相关产品推荐更多 >
万千商家帮你免费找货
0 人在求购买到急需产品
- 详细信息
- 文献和实验
- 技术资料
- 保存条件：
-20
- 保质期：
2年
- 英文名：
pESC-HIS
- 库存：
100
- 供应商：
上海烜雅生物科技有限公司
- 规格：
干粉/液体
基本信息
名称：pESC-HIS酿酒酵母质粒
别称: pESC-HIS
质粒属性
质粒简介
The pESC vectors are a series of epitope-tagging vectors for expression of eukaryotic genes in the yeast S. cerevisiae. Each vector contains GAL1 and GAL10 yeast promoters in opposing orientations. With these vectors you can introduce one or two cloned genes into a yeast host strain under the control of an inducible promoter. These vectors also feature an extensive polylinker sequence and the ability to generate end-specific RNA transcripts from T3 and T7 promoters. Each of the pESC vectors contains one of four different yeast-selectable markers (HIS3, TRP1, LEU2, or URA3) in the same vector backbone.
The pESC vectors contain DNA sequences coding for epitope peptides that can be specifically recognized by monoclonal antibodies. A sequence for the FLAG® epitope (DYKDDDDK) is located in the multiple cloning site (MCS) downstream of the GAL10 promoter; a sequence for the c-Myc epitope (EQKLISEEDL) is located in the MCS downstream of the GAL1 promoter. You can insert your gene of interest in front of the epitope sequence to generate C-terminal tagging or after the epitope sequence for N-terminal tagging. These tags allow the protein of interest to be studied without generating a specific antibody to that protein. The epitope-tagged fusion proteins can be studied in transformed cells using well-characterized antibodies.
质粒图谱
质粒序列
LOCUS Exported 6706 bp ds-DNA circular SYN 13-APR-2017
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pESC-HIS
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6706)
AUTHORS .
TITLE Direct Submission
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..6706
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 317..503
/gene="S. cerevisiae HIS3"
/note="HIS3 promoter"
CDS 504..1163
/codon_start=1
/gene="S. cerevisiae HIS3"
/product="imidazoleglycerol-phosphate dehydratase, required
for histidine biosynthesis"
/note="HIS3"
/note="yeast auxotrophic marker"
/translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
rep_origin complement(1423..1878)
/direction=LEFT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
terminator 1961..2126
/gene="S. cerevisiae ADH1"
/note="ADH1 terminator"
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
CDS complement(2274..2297)
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/note="FLAG"
/translation="DYKDDDDK"
promoter 2344..3008
/gene="GAL1 and GAL10 genes of S. cerevisiae"
/note="GAL1,10 promoter"
/note="divergent inducible promoter, regulated by Gal4"
protein_bind 2560..2677
/bound_moiety="Gal4"
/note="UAS"
/note="upstream activating sequence mediating
Gal4-dependent induction"
promoter 3019..3037
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 3055..3084
/codon_start=1
/product="Myc (human c-Myc oncogene) epitope tag"
/note="Myc"
/translation="EQKLISEEDL"
terminator 3114..3303
/gene="S. cerevisiae CYC1"
/note="CYC1 terminator"
/note="transcription terminator for CYC1"
rep_origin complement(3546..4134)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4305..5165)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5166..5270)
/gene="bla"
/note="AmpR promoter"
rep_origin 5297..6639
/note="2u ori"
/note="yeast 2u plasmid origin of replication"
protein_bind complement(6260..6307)
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FRT"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
ORIGIN
1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
181 accataaatt cccgttttaa gagcttggtg agcgctagga gtcactgcca ggtatcgttt
241 gaacacggca ttagtcaggg aagtcataac acagtccttt cccgcaattt tctttttcta
301 ttactcttgg cctcctctag tacactctat atttttttat gcctcggtaa tgattttcat
361 tttttttttt cccctagcgg atgactcttt ttttttctta gcgattggca ttatcacata
421 atgaattata cattatataa agtaatgtga tttcttcgaa gaatatacta aaaaatgagc
481 aggcaagata aacgaaggca aagatgacag agcagaaagc cctagtaaag cgtattacaa
541 atgaaaccaa gattcagatt gcgatctctt taaagggtgg tcccctagcg atagagcact
601 cgatcttccc agaaaaagag gcagaagcag tagcagaaca ggccacacaa tcgcaagtga
661 ttaacgtcca cacaggtata gggtttctgg accatatgat acatgctctg gccaagcatt
721 ccggctggtc gctaatcgtt gagtgcattg gtgacttaca catagacgac catcacacca
781 ctgaagactg cgggattgct ctcggtcaag cttttaaaga ggccctactg gcgcgtggag
841 taaaaaggtt tggatcagga tttgcgcctt tggatgaggc actttccaga gcggtggtag
901 atctttcgaa caggccgtac gcagttgtcg aacttggttt gcaaagggag aaagtaggag
961 atctctcttg cgagatgatc ccgcattttc ttgaaagctt tgcagaggct agcagaatta
1021 ccctccacgt tgattgtctg cgaggcaaga atgatcatca ccgtagtgag agtgcgttca
1081 aggctcttgc ggttgccata agagaagcca cctcgcccaa tggtaccaac gatgttccct
1141 ccaccaaagg tgttcttatg tagtgacacc gattatttaa agctgcagca tacgatatat
1201 atacatgtgt atatatgtat acctatgaat gtcagtaagt atgtatacga acagtatgat
1261 actgaagatg acaaggtaat gcatcattct atacgtgtca ttctgaacga ggcgcgcttt
1321 ccttttttct ttttgctttt tctttttttt tctcttgaac tcgacggatc tatgcggtgt
1381 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta aacgttaata
1441 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
1501 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
1561 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
1621 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
1681 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
1741 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
1801 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
1861 cgccgctaca gggcgcgtcg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
1921 gatcggtgcg ggcctcttcg ctattacgcc agctgaattg gagcgacctc atgctatacc
1981 tgagaaagca acctgaccta caggaaagag ttactcaaga ataagaattt tcgttttaaa
2041 acctaagagt cactttaaaa tttgtataca cttatttttt ttataactta tttaataata
2101 aaaatcataa atcataagaa attcgcttat ttagaagtgt caacaacgta tctaccaacg
2161 atttgaccct tttccatctt ttcgtaaatt tctggcaagg tagacaagcc gacaaccttg
2221 attggagact tgaccaaacc tctggcgaag aattgttaat taagagctca gatcttatcg
2281 tcgtcatcct tgtaatccat cgatactagt gcggccgccc tttagtgagg gttgaattcg
2341 aattttcaaa aattcttact ttttttttgg atggacgcaa agaagtttaa taatcatatt
2401 acatggcatt accaccatat acatatccat atacatatcc atatctaatc ttacttatat
2461 gttgtggaaa tgtaaagagc cccattatct tagcctaaaa aaaccttctc tttggaactt
2521 tcagtaatac gcttaactgc tcattgctat attgaagtac ggattagaag ccgccgagcg
2581 ggtgacagcc ctccgaagga agactctcct ccgtgcgtcc tcgtcttcac cggtcgcgtt
2641 cctgaaacgc agatgtgcct cgcgccgcac tgctccgaac aataaagatt ctacaatact
2701 agcttttatg gttatgaaga ggaaaaattg gcagtaacct ggccccacaa accttcaaat
2761 gaacgaatca aattaacaac cataggatga taatgcgatt agttttttag ccttatttct
2821 ggggtaatta atcagcgaag cgatgatttt tgatctatta acagatatat aaatgcaaaa
2881 actgcataac cactttaact aatactttca acattttcgg tttgtattac ttcttattca
2941 aatgtaataa aagtatcaac aaaaaattgt taatatacct ctatacttta acgtcaagga
3001 gaaaaaaccc cggatccgta atacgactca ctatagggcc cgggcgtcga catggaacag
3061 aagttgattt ccgaagaaga cctcgagtaa gcttggtacc gcggctagct aagatccgct
3121 ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata
3181 gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga
3241 cgcgtgtacg catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg
3301 aagatccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
3361 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
3421 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
3481 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
3541 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
3601 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
3661 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
3721 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
3781 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
3841 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
3901 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
3961 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
4021 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
4081 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
4141 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
4201 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
4261 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
4321 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
4381 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
4441 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
4501 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
4561 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
4621 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
4681 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
4741 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
4801 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
4861 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
4921 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
4981 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
5041 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
5101 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
5161 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
5221 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
5281 cgaaaagtgc cacctgaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc
5341 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa
5401 cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc
5461 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
5521 tgcaacgcga gagcgctatt ttaccaacaa agaatctata cttctttttt gttctacaaa
5581 aatgcatccc gagagcgcta tttttctaac aaagcatctt agattacttt ttttctcctt
5641 tgtgcgctct ataatgcagt ctcttgataa ctttttgcac tgtaggtccg ttaaggttag
5701 aagaaggcta ctttggtgtc tattttctct tccataaaaa aagcctgact ccacttcccg
5761 cgtttactga ttactagcga agctgcgggt gcattttttc aagataaagg catccccgat
5821 tatattctat accgatgtgg attgcgcata ctttgtgaac agaaagtgat agcgttgatg
5881 attcttcatt ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta tatactacgt
5941 ataggaaatg tttacatttt cgtattgttt tcgattcact ctatgaatag ttcttactac
6001 aatttttttg tctaaagagt aatactagag ataaacataa aaaatgtaga ggtcgagttt
6061 agatgcaagt tcaaggagcg aaaggtggat gggtaggtta tatagggata tagcacagag
6121 atatatagca aagagatact tttgagcaat gtttgtggaa gcggtattcg caatatttta
6181 gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa gtgcgtcttc agagcgcttt
6241 tggttttcaa aagcgctctg aagttcctat actttctaga gaataggaac ttcggaatag
6301 gaacttcaaa gcgtttccga aaacgagcgc ttccgaaaat gcaacgcgag ctgcgcacat
6361 acagctcact gttcacgtcg cacctatatc tgcgtgttgc ctgtatatat atatacatga
6421 gaagaacggc atagtgcgtg tttatgctta aatgcgtact tatatgcgtc tatttatgta
6481 ggatgaaagg tagtctagta cctcctgtga tattatccca ttccatgcgg ggtatcgtat
6541 gcttccttca gcactaccct ttagctgttc tatatgctgc cactcctcaa ttggattagt
6601 ctcatccttc aatgctatca tttcctttga tattggatca tctaagaaac cattattatc
6661 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc
//
质粒菌株产品操作说明书
一、扩增流程
收到产品后，请先根据产品管壁标签来判断产品形式，并在扩增前准确查找该质粒菌株的抗性、感受态和培养温度。
1、质粒干粉（常温运输，存于-20度，90天保质期，请务必转化挑单克隆培养，不要直接使用和测序）
①收到质粒干粉后请先5000rpm离心1min，再加入20μl ddH2O去离子水溶解质粒；
②取1支100μl 感受态于冰上解冻10min，加入2μl质粒，再冰浴30min后，42℃热激60s，不要搅动，再冰浴2min；（从第二步开始均要在超净工作台中无菌操作）
③加入900μl无抗的LB液体培养基，180rpm震荡37℃培养45min；
④6000rpm离心5min，仅留100μl上清液重悬细菌沉淀，并涂布至目标质粒抗性的LB平板上；（可使用本平台的平板涂布专用玻璃珠进行涂布，可以比传统涂布方法获得更多转化子）
⑤将平板正向培养1h，再倒置37℃培养14h。如果要求是30度则培养20h；
（菌落过多则将质粒稀释后再转化。没有菌落则加入10μl质粒转化。另不要直接转表达感受态，要先转克隆感受态，重提质粒后再导入表达感受态）
⑥挑取单菌落至LB液体培养基中，加入对应抗生素，220rpm震荡培养14h，根据实验需要和质粒提取试剂盒说明书提取质粒。
2、甘油菌种（冰袋运输，存于-80℃，保质期90天，请务必划线挑单克隆培养）
四区划线后挑单菌落培养，酵母菌需要先液体复苏再四区划线，再挑单菌落液体培养。
3、穿刺菌种（冰袋运输，存于4℃，保质期7天）
穿刺接种，液体培养后四区划线，再挑单菌落液体培养。
4、菌落平板（冰袋运输，存于4℃，保质期7天）
直接挑取单菌落至液体培养基中。
5、液体质粒（冰袋运输，存于-20℃，保质期90天）
单独提取的液体质粒收到后可直接使用。
6、滤纸质粒（常温运输，存于-20度，90天保质期，请务必转化挑单克隆培养，不要直接使用和测序）
收到货后将滤纸画圈部分剪下放入EP管中，加100ul无菌水将滤纸浸湿并浸泡5min，吸取5ul质粒转化，离心全涂。
二、转化图片
名称：pESC-HIS酿酒酵母质粒
别称: pESC-HIS
| 启动子: | GAL1,10 |
|---|---|
| 复制子: | 2U |
| 质粒大小: | 6706bp |
| 原核抗性: | Amp |
| 筛选标记: | HIS4 |
| 克隆菌株: | DH5a |
| 培养条件: | 37度 |
质粒属性
| 载体宿主： | 酵母菌 |
|---|---|
| 载体用途： | 蛋白表达 |
| 基因种属： | 多框空载 |
| 基因类型： | ORF |
| 原核抗性： | Amp |
| 筛选标记： | |
| 荧光蛋白： |
质粒简介
The pESC vectors are a series of epitope-tagging vectors for expression of eukaryotic genes in the yeast S. cerevisiae. Each vector contains GAL1 and GAL10 yeast promoters in opposing orientations. With these vectors you can introduce one or two cloned genes into a yeast host strain under the control of an inducible promoter. These vectors also feature an extensive polylinker sequence and the ability to generate end-specific RNA transcripts from T3 and T7 promoters. Each of the pESC vectors contains one of four different yeast-selectable markers (HIS3, TRP1, LEU2, or URA3) in the same vector backbone.
The pESC vectors contain DNA sequences coding for epitope peptides that can be specifically recognized by monoclonal antibodies. A sequence for the FLAG® epitope (DYKDDDDK) is located in the multiple cloning site (MCS) downstream of the GAL10 promoter; a sequence for the c-Myc epitope (EQKLISEEDL) is located in the MCS downstream of the GAL1 promoter. You can insert your gene of interest in front of the epitope sequence to generate C-terminal tagging or after the epitope sequence for N-terminal tagging. These tags allow the protein of interest to be studied without generating a specific antibody to that protein. The epitope-tagged fusion proteins can be studied in transformed cells using well-characterized antibodies.
质粒图谱
质粒序列
LOCUS Exported 6706 bp ds-DNA circular SYN 13-APR-2017
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pESC-HIS
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6706)
AUTHORS .
TITLE Direct Submission
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..6706
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 317..503
/gene="S. cerevisiae HIS3"
/note="HIS3 promoter"
CDS 504..1163
/codon_start=1
/gene="S. cerevisiae HIS3"
/product="imidazoleglycerol-phosphate dehydratase, required
for histidine biosynthesis"
/note="HIS3"
/note="yeast auxotrophic marker"
/translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
rep_origin complement(1423..1878)
/direction=LEFT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
terminator 1961..2126
/gene="S. cerevisiae ADH1"
/note="ADH1 terminator"
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
CDS complement(2274..2297)
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/note="FLAG"
/translation="DYKDDDDK"
promoter 2344..3008
/gene="GAL1 and GAL10 genes of S. cerevisiae"
/note="GAL1,10 promoter"
/note="divergent inducible promoter, regulated by Gal4"
protein_bind 2560..2677
/bound_moiety="Gal4"
/note="UAS"
/note="upstream activating sequence mediating
Gal4-dependent induction"
promoter 3019..3037
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 3055..3084
/codon_start=1
/product="Myc (human c-Myc oncogene) epitope tag"
/note="Myc"
/translation="EQKLISEEDL"
terminator 3114..3303
/gene="S. cerevisiae CYC1"
/note="CYC1 terminator"
/note="transcription terminator for CYC1"
rep_origin complement(3546..4134)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4305..5165)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5166..5270)
/gene="bla"
/note="AmpR promoter"
rep_origin 5297..6639
/note="2u ori"
/note="yeast 2u plasmid origin of replication"
protein_bind complement(6260..6307)
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FRT"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
ORIGIN
1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
181 accataaatt cccgttttaa gagcttggtg agcgctagga gtcactgcca ggtatcgttt
241 gaacacggca ttagtcaggg aagtcataac acagtccttt cccgcaattt tctttttcta
301 ttactcttgg cctcctctag tacactctat atttttttat gcctcggtaa tgattttcat
361 tttttttttt cccctagcgg atgactcttt ttttttctta gcgattggca ttatcacata
421 atgaattata cattatataa agtaatgtga tttcttcgaa gaatatacta aaaaatgagc
481 aggcaagata aacgaaggca aagatgacag agcagaaagc cctagtaaag cgtattacaa
541 atgaaaccaa gattcagatt gcgatctctt taaagggtgg tcccctagcg atagagcact
601 cgatcttccc agaaaaagag gcagaagcag tagcagaaca ggccacacaa tcgcaagtga
661 ttaacgtcca cacaggtata gggtttctgg accatatgat acatgctctg gccaagcatt
721 ccggctggtc gctaatcgtt gagtgcattg gtgacttaca catagacgac catcacacca
781 ctgaagactg cgggattgct ctcggtcaag cttttaaaga ggccctactg gcgcgtggag
841 taaaaaggtt tggatcagga tttgcgcctt tggatgaggc actttccaga gcggtggtag
901 atctttcgaa caggccgtac gcagttgtcg aacttggttt gcaaagggag aaagtaggag
961 atctctcttg cgagatgatc ccgcattttc ttgaaagctt tgcagaggct agcagaatta
1021 ccctccacgt tgattgtctg cgaggcaaga atgatcatca ccgtagtgag agtgcgttca
1081 aggctcttgc ggttgccata agagaagcca cctcgcccaa tggtaccaac gatgttccct
1141 ccaccaaagg tgttcttatg tagtgacacc gattatttaa agctgcagca tacgatatat
1201 atacatgtgt atatatgtat acctatgaat gtcagtaagt atgtatacga acagtatgat
1261 actgaagatg acaaggtaat gcatcattct atacgtgtca ttctgaacga ggcgcgcttt
1321 ccttttttct ttttgctttt tctttttttt tctcttgaac tcgacggatc tatgcggtgt
1381 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta aacgttaata
1441 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
1501 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
1561 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
1621 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
1681 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
1741 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
1801 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
1861 cgccgctaca gggcgcgtcg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
1921 gatcggtgcg ggcctcttcg ctattacgcc agctgaattg gagcgacctc atgctatacc
1981 tgagaaagca acctgaccta caggaaagag ttactcaaga ataagaattt tcgttttaaa
2041 acctaagagt cactttaaaa tttgtataca cttatttttt ttataactta tttaataata
2101 aaaatcataa atcataagaa attcgcttat ttagaagtgt caacaacgta tctaccaacg
2161 atttgaccct tttccatctt ttcgtaaatt tctggcaagg tagacaagcc gacaaccttg
2221 attggagact tgaccaaacc tctggcgaag aattgttaat taagagctca gatcttatcg
2281 tcgtcatcct tgtaatccat cgatactagt gcggccgccc tttagtgagg gttgaattcg
2341 aattttcaaa aattcttact ttttttttgg atggacgcaa agaagtttaa taatcatatt
2401 acatggcatt accaccatat acatatccat atacatatcc atatctaatc ttacttatat
2461 gttgtggaaa tgtaaagagc cccattatct tagcctaaaa aaaccttctc tttggaactt
2521 tcagtaatac gcttaactgc tcattgctat attgaagtac ggattagaag ccgccgagcg
2581 ggtgacagcc ctccgaagga agactctcct ccgtgcgtcc tcgtcttcac cggtcgcgtt
2641 cctgaaacgc agatgtgcct cgcgccgcac tgctccgaac aataaagatt ctacaatact
2701 agcttttatg gttatgaaga ggaaaaattg gcagtaacct ggccccacaa accttcaaat
2761 gaacgaatca aattaacaac cataggatga taatgcgatt agttttttag ccttatttct
2821 ggggtaatta atcagcgaag cgatgatttt tgatctatta acagatatat aaatgcaaaa
2881 actgcataac cactttaact aatactttca acattttcgg tttgtattac ttcttattca
2941 aatgtaataa aagtatcaac aaaaaattgt taatatacct ctatacttta acgtcaagga
3001 gaaaaaaccc cggatccgta atacgactca ctatagggcc cgggcgtcga catggaacag
3061 aagttgattt ccgaagaaga cctcgagtaa gcttggtacc gcggctagct aagatccgct
3121 ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata
3181 gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga
3241 cgcgtgtacg catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg
3301 aagatccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
3361 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
3421 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
3481 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
3541 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
3601 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
3661 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
3721 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
3781 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
3841 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
3901 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
3961 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
4021 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
4081 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
4141 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
4201 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
4261 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
4321 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
4381 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
4441 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
4501 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
4561 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
4621 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
4681 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
4741 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
4801 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
4861 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
4921 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
4981 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
5041 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
5101 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
5161 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
5221 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
5281 cgaaaagtgc cacctgaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc
5341 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa
5401 cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc
5461 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
5521 tgcaacgcga gagcgctatt ttaccaacaa agaatctata cttctttttt gttctacaaa
5581 aatgcatccc gagagcgcta tttttctaac aaagcatctt agattacttt ttttctcctt
5641 tgtgcgctct ataatgcagt ctcttgataa ctttttgcac tgtaggtccg ttaaggttag
5701 aagaaggcta ctttggtgtc tattttctct tccataaaaa aagcctgact ccacttcccg
5761 cgtttactga ttactagcga agctgcgggt gcattttttc aagataaagg catccccgat
5821 tatattctat accgatgtgg attgcgcata ctttgtgaac agaaagtgat agcgttgatg
5881 attcttcatt ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta tatactacgt
5941 ataggaaatg tttacatttt cgtattgttt tcgattcact ctatgaatag ttcttactac
6001 aatttttttg tctaaagagt aatactagag ataaacataa aaaatgtaga ggtcgagttt
6061 agatgcaagt tcaaggagcg aaaggtggat gggtaggtta tatagggata tagcacagag
6121 atatatagca aagagatact tttgagcaat gtttgtggaa gcggtattcg caatatttta
6181 gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa gtgcgtcttc agagcgcttt
6241 tggttttcaa aagcgctctg aagttcctat actttctaga gaataggaac ttcggaatag
6301 gaacttcaaa gcgtttccga aaacgagcgc ttccgaaaat gcaacgcgag ctgcgcacat
6361 acagctcact gttcacgtcg cacctatatc tgcgtgttgc ctgtatatat atatacatga
6421 gaagaacggc atagtgcgtg tttatgctta aatgcgtact tatatgcgtc tatttatgta
6481 ggatgaaagg tagtctagta cctcctgtga tattatccca ttccatgcgg ggtatcgtat
6541 gcttccttca gcactaccct ttagctgttc tatatgctgc cactcctcaa ttggattagt
6601 ctcatccttc aatgctatca tttcctttga tattggatca tctaagaaac cattattatc
6661 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc
//
质粒菌株产品操作说明书
一、扩增流程
收到产品后，请先根据产品管壁标签来判断产品形式，并在扩增前准确查找该质粒菌株的抗性、感受态和培养温度。
1、质粒干粉（常温运输，存于-20度，90天保质期，请务必转化挑单克隆培养，不要直接使用和测序）
①收到质粒干粉后请先5000rpm离心1min，再加入20μl ddH2O去离子水溶解质粒；
②取1支100μl 感受态于冰上解冻10min，加入2μl质粒，再冰浴30min后，42℃热激60s，不要搅动，再冰浴2min；（从第二步开始均要在超净工作台中无菌操作）
③加入900μl无抗的LB液体培养基，180rpm震荡37℃培养45min；
④6000rpm离心5min，仅留100μl上清液重悬细菌沉淀，并涂布至目标质粒抗性的LB平板上；（可使用本平台的平板涂布专用玻璃珠进行涂布，可以比传统涂布方法获得更多转化子）
⑤将平板正向培养1h，再倒置37℃培养14h。如果要求是30度则培养20h；
（菌落过多则将质粒稀释后再转化。没有菌落则加入10μl质粒转化。另不要直接转表达感受态，要先转克隆感受态，重提质粒后再导入表达感受态）
⑥挑取单菌落至LB液体培养基中，加入对应抗生素，220rpm震荡培养14h，根据实验需要和质粒提取试剂盒说明书提取质粒。
2、甘油菌种（冰袋运输，存于-80℃，保质期90天，请务必划线挑单克隆培养）
四区划线后挑单菌落培养，酵母菌需要先液体复苏再四区划线，再挑单菌落液体培养。
3、穿刺菌种（冰袋运输，存于4℃，保质期7天）
穿刺接种，液体培养后四区划线，再挑单菌落液体培养。
4、菌落平板（冰袋运输，存于4℃，保质期7天）
直接挑取单菌落至液体培养基中。
5、液体质粒（冰袋运输，存于-20℃，保质期90天）
单独提取的液体质粒收到后可直接使用。
6、滤纸质粒（常温运输，存于-20度，90天保质期，请务必转化挑单克隆培养，不要直接使用和测序）
收到货后将滤纸画圈部分剪下放入EP管中，加100ul无菌水将滤纸浸湿并浸泡5min，吸取5ul质粒转化，离心全涂。
二、转化图片
| P0506/pPICZαB毕赤酵母质粒 |
| P0239/pPICZαC毕赤酵母质粒 |
| P0217/pGAPZA毕赤酵母质粒 |
| P0218/pGAPZB毕赤酵母质粒 |
| P0219/pGAPZC毕赤酵母质粒 |
| P0220/pGAPZαA毕赤酵母质粒 |
| P1231/pGAPZαB毕赤酵母质粒 |
| P1232/pGAPZαC毕赤酵母质粒 |
| P4527/pBGP1-Zeo毕赤酵母独立自复制质粒 |
| P0221/pPinkα-HC毕赤酵母质粒 |
| P0404/pPINK-LC毕赤酵母质粒 |
风险提示：丁香通仅作为第三方平台，为商家信息发布提供平台空间。用户咨询产品时请注意保护个人信息及财产安全，合理判断，谨慎选购商品，商家和用户对交易行为负责。对于医疗器械类产品，请先查证核实企业经营资质和医疗器械产品注册证情况。
文献和实验相关实验
相关专题 完美酵母质粒提取实验Protocol 问：请教如何从酿酒酵母抽提质粒 ? 用胞壁松解酶处理以后，再用SDS处理，是否就可以破碎酵母了，因为是抽提质粒，不是抽基因组 DNA，可以不用蛋白酶K处理吗?请各位朋友提宝贵建议(不用玻璃珠破细胞)。我要尽可能把样品中的质粒都抽提出来，然后 REAL TIME PCR定量，好计算每个酵母细胞中的质粒拷贝数目。因为样品少，不能做SOUTHERN BOLT计算拷贝。谢谢
相关专题 酶联免疫斑点法（ELISpot） 酵母细胞的细胞壁比较厚，不容易破壁，不如大肠杆菌的质粒 容易提取，最近做了些酿酒酵母的实验，从酿酒酵母中提取质粒，现在就总结下实验的方法和步骤。 酵母细胞质粒 提取步骤 1. 接种单菌落(待检测酵母细胞)于25mLYNB(补加氨基酸营养物)培养基中，30℃振荡培养过夜。 2.第二天取一滴菌液于进行显微镜下观察,目镜用16,物镜用40倍观察细胞壁破碎前
pYES2.0 酵母质粒 Amp 酿酒酵母载体 pRS403 酵母质粒 pRS423 酵母质粒 pRC426gal 酵母质粒
pESC-HIS酿酒酵母质粒
￥100 - 1000
