| 质粒名称: | pWZL Blast Snail ER |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 18798 |
| 存储实验室: | Bob Weinberg |
|
载体骨架基本信息 |
|
| 载体名称: | pWZL-Blast |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5100 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，逆转录病毒 |
| 筛选标记: | Blasticidin |
| 高拷贝/低拷贝: | 未知 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | snail |
| 物种来源: | H. sapiens (human) |
| 插入大小: | -- |
| 基因突变: | 内质网G525R突变。克隆的ER序列中G525R点突变的存在废除了它与17β-雌二醇的结合，同时保持其对合成配体4'-羟基他莫昔芬（4OHT）的完全敏感性。 |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | 内质网激素结合域（插入端C端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BamHI (未破坏) |
| 3' 克隆位点: | BamHI (在克隆过程中销毁) |
| 5' 测序引物 | pBABE 5' |
| 3' 测序引物 | IRES-R (5'-GCATTCCTTTGGCGAGAG-3') |
|
文献引用方法 |
|
| 材料和方法部分: | pWZL Blast Snail ER was a gift from Bob Weinberg (Addgene plasmid # 18798 ; http://n2t.net/addgene:18798 ; RRID:Addgene_18798) |
| 参考文献部分: | The epithelial-mesenchymal transition generates cells with properties of stem cells. Mani SA, Guo W, Liao MJ, Eaton EN, Ayyanan A, Zhou AY, Brooks M, Reinhard F, Zhang CC, Shipitsin M, Campbell LL, Polyak K, Brisken C, Yang J, Weinberg RA. Cell. 2008 May 16. 133(4):704-15. 10.1016/j.cell.2008.03.027 PubMed 18485877 |
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 pWZL Blast Snail ER 6814 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pWZL Blast Snail ER
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6814)
AUTHORS Mani SA, Guo W, Liao MJ, Eaton EN, Ayyanan A, Zhou AY, Brooks M,
Reinhard F, Zhang CC, Shipitsin M, Campbell LL, Polyak K, Brisken C,
Yang J, Weinberg RA
TITLE The epithelial-mesenchymal transition generates cells with
properties of stem cells.
JOURNAL Cell. 2008 May 16. 133(4):704-15.
PUBMED 18485877
REFERENCE 2 (bases 1 to 6814)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6814
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 8..477
/note="long terminal repeat from Moloney murine leukemia
virus"
misc_feature 542..741
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 942..1358
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 1288..1310
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 1326..1342
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
misc_feature 3231..3692
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(3287..3304)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 3514..3533
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
LTR 4308..4777
/note="long terminal repeat from Moloney murine leukemia
virus"
primer_bind complement(4907..4924)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5078..5666)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5158..5177)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5826..6686)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 6449..6468
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6687..6791)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
541 caagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt
901 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
961 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
1021 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
1081 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
1141 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
1201 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
1261 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
1321 cctcccttta tccagccctc actccttctc taggcgccgg ccggatccat gccgcgctct
1381 ttcctcgtca ggaagccctc cgaccccaat cggaagccta actacagcga gctgcaggac
1441 tctaatccag agtttacctt ccagcagccc tacgaccagg cccacctgct ggcagccatc
1501 ccacctccgg agatcctcaa ccccaccgcc tcgctgccaa tgctcatctg ggactctgtc
1561 ctggcgcccc aagcccagcc aattgcctgg gcctcccttc ggctccagga gagtcccagg
1621 gtggcagagc tgacctccct gtcagatgag gacagtggga aaggctccca gccccccagc
1681 ccaccctcac cggctccttc gtccttctcc tctacttcag tctcttcctt ggaggccgag
1741 gcctatgctg ccttcccagg cttgggccaa gtgcccaagc agctggccca gctctctgag
1801 gccaaggatc tccaggctcg aaaggccttc aactgcaaat actgcaacaa ggaatacctc
1861 agcctgggtg ccctcaagat gcacatccga agccacacgc tgccctgcgt ctgcggaacc
1921 tgcgggaagg ccttctctag gccctggctg ctacaaggcc atgtccggac ccacactggc
1981 gagaagccct tctcctgtcc ccactgcagc cgtgccttcg ctgaccgctc caacctgcgg
2041 gcccacctcc agacccactc agatgtcaag aagtaccagt gccaggcgtg tgctcggacc
2101 ttctcccgaa tgtccctgct ccacaagcac caagagtccg gctgctcagg atgtccccgc
2161 gcggatccac gaaatgaaat gggtgcttca ggagacatga gggctgccaa cctttggcca
2221 agccctcttg tgattaagca cactaagaag aatagccctg ccttgtcctt gacagctgac
2281 cagatggtca gtgccttgtt ggatgctgaa ccgcccatga tctattctga atatgatcct
2341 tctagaccct tcagtgaagc ctcaatgatg ggcttattga ccaacctagc agatagggag
2401 ctggttcata tgatcaactg ggcaaagaga gtgccaggct ttggggactt gaatctccat
2461 gatcaggtcc accttctcga gtgtgcctgg ctggagattc tgatgattgg tctcgtctgg
2521 cgctccatgg aacacccggg gaagctcctg tttgctccta acttgctcct ggacaggaat
2581 caaggtaaat gtgtggaagg catggtggag atctttgaca tgttgcttgc tacgtcaagt
2641 cggttccgca tgatgaacct gcagggtgaa gagtttgtgt gcctcaaatc catcattttg
2701 cttaattccg gagtgtacac gtttctgtcc agcaccttga agtctctgga agagaaggac
2761 cacatccacc gtgtcctgga caagatcaca gacactttga tccacctgat ggccaaagct
2821 ggcctgactc tgcagcagca gcatcgccgc ctagctcagc tccttctcat tctttcccat
2881 atccggcata tgagtaacaa acgcatggag catctctaca acatgaaatg caagaacgtg
2941 gtacccctct atgacctgct cctggagatg ttggatgccc accgccttca tgccccagcc
3001 agtcgcatgg gagtgccccc agaggagccc agccagaccc agctggccac caccagctcc
3061 acttcagcac attccttaca aacctactac atacccccgg aagcagaggg cttccccaac
3121 acgatctgag agctcctact agtgaattca ctcgagatac gtaaaacaat tgaagatcta
3181 gtcgaccaat tccggttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg
3241 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga
3301 atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa
3361 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc
3421 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac
3481 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag
3541 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc
3601 acatgcttta catgtgttta gtcgaggtta aaaaacgtct aggccccccg aaccacgggg
3661 acgtggtttt cctttgaaaa acacgatgat aataccatga aaacatttaa catttctcaa
3721 caggatctag aattagtaga agtagcgaca gagaagatta caatgcttta tgaggataat
3781 aaacatcatg tgggagcggc aattcgtacg aaaacaggag aaatcatttc ggcagtacat
3841 attgaagcgt atataggacg agtaactgtt tgtgcagaag ccattgcgat tggtagtgca
3901 gtttcgaatg gacaaaagga ttttgacacg attgtagctg ttagacaccc ttattctgac
3961 gaagtagata gaagtattcg agtggtaagt ccttgtggta tgtgtaggga gttgatttca
4021 gactatgcac cagattgttt tgtgttaata gaaatgaatg gcaagttagt caaaactacg
4081 attgaagaac tcattccact caaatatacc cgaaattaaa agttttacca taccaagctc
4141 atcgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac
4201 ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata
4261 actgagaata gagaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc
4321 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac
4381 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
4441 aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat
4501 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
4561 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
4621 acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc
4681 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
4741 tctgagtgat tgactacccg tcagcggggg tctttcacat gcagcatgta tcaaaattaa
4801 tttggttttt tttcttaagt atttacatta aatggccata gttgcattaa tgaatcggcc
4861 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
4921 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
4981 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
5041 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
5101 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
5161 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
5221 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
5281 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
5341 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
5401 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
5461 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
5521 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
5581 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
5641 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
5701 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
5761 tcacctagat ccttttgcgg ccgcaaatca atctaaagta tatatgagta aacttggtct
5821 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
5881 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
5941 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
6001 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
6061 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
6121 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
6181 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
6241 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
6301 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
6361 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
6421 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
6481 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
6541 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
6601 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
6661 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
6721 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
6781 ggggttccgc gcacatttcc ccgaaaagtg ccac
//
