| 质粒名称: | FUW-M2rtTA |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 20342 |
| 存储实验室: | Rudolf Jaenisch |
|
载体骨架基本信息 |
|
| 载体名称: | FUW (Lenti-Lox-Ubi) |
| 载体出品公司: | homemade |
| 空载体大小: | 7231 bp |
| 完整质粒大小: | 7979 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | Stbl3 |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | M2rtTA |
| 物种来源: | bacteria |
| 插入大小: | 792 bp |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | EcoRI(未破坏) |
| 3’克隆位点: | EcoRI(未破坏) |
| 5’测序引物 | n/a |
|
文献引用方法 |
|
| 材料和方法部分: | FUW-M2rtTA was a gift from Rudolf Jaenisch (Addgene plasmid # 20342 ; http://n2t.net/addgene:20342 ; RRID:Addgene_20342) |
| 参考文献部分: | A drug-inducible system for direct reprogramming of human somatic cells to pluripotency. Hockemeyer D, Soldner F, Cook EG, Gao Q, Mitalipova M, Jaenisch R. Cell Stem Cell. 2008 Sep 11. 3(3):346-53. 10.1016/j.stem.2008.08.014 PubMed 18786421 |
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 FUW-M2rtTA 7979 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Lentiviral plasmid expressing the reverse tetracycline
transactivator.
ACCESSION .
VERSION .
KEYWORDS FUW-M2rtTA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7979)
AUTHORS Hockemeyer D, Soldner F, Cook EG, Gao Q, Mitalipova M, Jaenisch R
TITLE A drug-inducible system for direct reprogramming of human somatic
cells to pluripotency.
JOURNAL Cell Stem Cell. 2008 Sep 11. 3(3):346-53.
PUBMED 18786421
REFERENCE 2 (bases 1 to 7979)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7979
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(46..65)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 237..616
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 618..816
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 771..791
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 834..1014
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1061..1186
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1679..1912
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 2097..2141
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 2439..2556
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2621..3832
/label=UbC promoter
/note="human ubiquitin C promoter"
primer_bind 3669..3688
/label=hUBCpro-F
/note="Human Ubiquitin C (UbC) promoter, forward primer"
CDS 3861..4607
/codon_start=1
/product="improved tetracycline-controlled transactivator"
/label=rtTA-Advanced
/note="In the Tet-On(R) system, rtTA-Advanced binds to the
Tet-responsive element and stimulates transcription only in
the presence of tetracycline or doxycycline."
/translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
DALDDFDLDMLPADALDDFDLDMLPG"
misc_feature 4644..5232
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4697..4717)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(5115..5126)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(5235..5251)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5236..5252)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 5757..5937
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
primer_bind complement(5963..5980)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5969..6080
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin complement(6226..6814)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6306..6325)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6985..7845)
/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 7608..7627
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7846..7950)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
781 cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc
841 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
901 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
961 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
1021 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
1081 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
1141 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
1201 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
1261 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct
1321 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
1381 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
1441 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
1501 aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga
1561 gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag
1621 tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag
1681 gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga
1741 cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc
1801 tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc
1861 tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt
1921 ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta
1981 ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta
2041 acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga
2101 atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa
2161 caaattggct gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa
2221 gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat
2281 cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag
2341 aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatcg gcactgcgtg
2401 cgccaattct gcagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt
2461 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
2521 gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga
2581 gatccagttt ggttaattaa cccgtgtcgg ctccagatct ggcctccgcg ccgggttttg
2641 gcgcctcccg cgggcgcccc cctcctcacg gcgagcgctg ccacgtcaga cgaagggcgc
2701 agcgagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg ctcataagac
2761 tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt gggtgactct
2821 agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc ccttctcggc
2881 gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa ggacgcgccg
2941 ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt gtttgtggat
3001 cgctgtgatc gtcacttggt gagtagcggg ctgctgggct ggccggggct ttcgtggccg
3061 ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc caagggctgt agtctgggtc
3121 cgcgagcaag gttgccctga actgggggtt ggggggagcg cagcaaaatg gcggctgttc
3181 ccgagtcttg aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg
3241 gggcatggtg ggcggcaaga acccaaggtc ttgaggcctt cgctaatgcg ggaaagctct
3301 tattcgggtg agatgggctg gggcaccatc tggggaccct gacgtgaagt ttgtcactga
3361 ctggagaact cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca
3421 gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt
3481 ggcttataat gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg
3541 caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg
3601 gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct
3661 taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga
3721 agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta
3781 gactagtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag acgaagcttg
3841 ggcccgggaa ttaattcacc atgtctagac tggacaagag caaagtcata aacggcgctc
3901 tggaattact caatggagtc ggtatcgaag gcctgacgac aaggaaactc gctcaaaagc
3961 tgggagttga gcagcctacc ctgtactggc acgtgaagaa caagcgggcc ctgctcgatg
4021 ccctgccaat cgagatgctg gacaggcatc atacccactt ctgccccctg gaaggcgagt
4081 catggcaaga ctttctgcgg aacaacgcca agtcattccg ctgtgctctc ctctcacatc
4141 gcgacggggc taaagtgcat ctcggcaccc gcccaacaga gaaacagtac gaaaccctgg
4201 aaaatcagct cgcgttcctg tgtcagcaag gcttctccct ggagaacgca ctgtacgctc
4261 tgtccgccgt gggccacttt acactgggct gcgtattgga ggaacaggag catcaagtag
4321 caaaagagga aagagagaca cctaccaccg attctatgcc cccacttctg agacaagcaa
4381 ttgagctgtt cgaccggcag ggagccgaac ctgccttcct tttcggcctg gaactaatca
4441 tatgtggcct ggagaaacag ctaaagtgcg aaagcggcgg gccggccgac gcccttgacg
4501 attttgactt agacatgctc ccagccgatg cccttgacga ctttgacctt gatatgctgc
4561 ctgctgacgc tcttgacgat tttgaccttg acatgctccc cgggtaacta agtaaggatc
4621 aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
4681 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
4741 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
4801 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
4861 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
4921 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
4981 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
5041 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
5101 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
5161 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
5221 ccgcctcccc gcatcgatac cgtcgacctc gagacctaga aaaacatgga gcaatcacaa
5281 gtagcaatac agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg
5341 aggtgggttt tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg
5401 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
5461 gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc cctgattggc
5521 agaactacac accagggcca gggatcagat atccactgac ctttggatgg tgctacaagc
5581 tagtaccagt tgagcaagag aaggtagaag aagccaatga aggagagaac acccgcttgt
5641 tacaccctgt gagcctgcat gggatggatg acccggagag agaagtatta gagtggaggt
5701 ttgacagccg cctagcattt catcacatgg cccgagagct gcatccggac tgtactgggt
5761 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
5821 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
5881 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcaggg
5941 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
6001 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
6061 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
6121 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgt gagcaaaagg
6181 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
6241 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
6301 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
6361 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
6421 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
6481 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
6541 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
6601 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
6661 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
6721 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
6781 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
6841 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
6901 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
6961 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
7021 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
7081 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
7141 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
7201 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
7261 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
7321 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
7381 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
7441 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
7501 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
7561 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
7621 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
7681 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
7741 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
7801 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
7861 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
7921 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacg
//
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