| 质粒名称: | pMXs-IP-mVenus-TOSI |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 172491 |
| 存储实验室: | David Scadden |
|
载体骨架基本信息 |
|
| 载体名称: | pMXs-IP |
| 载体出品公司: | Toshi Kitamura at University of Tokyo |
| 空载体大小: | 5800 bp |
| 完整质粒大小: | 6800 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，逆转录病毒 |
| 筛选标记: | 嘌呤霉素 |
| 载体抗性: | 氨苄青霉素，100μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Pdcd4 |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | 240 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | MLV LTR |
| 标签/融合蛋白: | mVenus（插入件上的N端子） |
| 克隆方法: | TOPO Cloning |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | CCACCGCCCTCAAAGTAGACG |
| 3’测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pMXs-IP-mVenus-TOSI was a gift from David Scadden (Addgene plasmid # 172491 ; http://n2t.net/addgene:172491 ; RRID:Addgene_172491) |
| 参考文献部分: | Imaging dynamic mTORC1 pathway activity in vivo reveals marked shifts that support time-specific inhibitor therapy in AML. Oki T, Mercier F, Kato H, Jung Y, McDonald TO, Spencer JA, Mazzola MC, van Gastel N, Lin CP, Michor F, Kitamura T, Scadden DT. Nat Commun. 2021 Jan 11;12(1):245. doi: 10.1038/s41467-020-20491-8. 10.1038/s41467-020-20491-8 PubMed 33431855 |
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 pMXs-IP-mVenus-TOSI 6825 bp ds-DNA circular SYN 28-OCT-2021
DEFINITION mVenus-TOSI (TOr-Signal-Indicator) is a fluorescent reporter for
mTORC1 signaling (monitoring PDCD4 degradation) which can be
introduced to mammalian cells by retrovirus vector. .
ACCESSION .
VERSION .
KEYWORDS pMXs-IP-mVenus-TOSI
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6825)
AUTHORS Oki T, Mercier F, Kato H, Jung Y, McDonald TO, Spencer JA, Mazzola
MC, van Gastel N, Lin CP, Michor F, Kitamura T, Scadden DT
TITLE Imaging dynamic mTORC1 pathway activity in vivo reveals marked
shifts that support time-specific inhibitor therapy in AML.
JOURNAL Nat Commun. 2021 Jan 11;12(1):245. doi: 10.1038/s41467-020-20491-8.
PUBMED 33431855
REFERENCE 2 (bases 1 to 6825)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Oct 28, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6825
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 38..54
/label=pBMN 5'
/note="MMLV sequence, for inserts in pBMN retroviral
vector"
CDS 129..845
/codon_start=1
/product="Venus YFP with monomerizing A206K mutation (Nagai
et al., 2002; Kremers et al., 2006)"
/label=mVenus
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(174..195)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(435..454)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 782..803
/label=EGFP-C
/note="EGFP, forward primer"
CDS 876..896
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
misc_feature 1176..1762
/label=IRES2
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(1343..1360)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 1570..1589
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 1763..2365
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MVTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind 2262..2282
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
LTR 2536..3005
/note="long terminal repeat from Moloney murine leukemia
virus"
primer_bind complement(3135..3152)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3306..3894)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3386..3405)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4058..4918)
/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 4681..4700
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4919..5023)
/gene="bla"
/label=AmpR promoter
LTR 5053..5646
/note="long terminal repeat from Moloney murine leukemia
virus"
misc_feature 5709..6066
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 6125..6541
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 6471..6493
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 6509..6525
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
misc_feature 6551..100
/label=pol region
/note="Moloney murine leukemia virus (MMLV) pol region
containing the splice acceptor site"
ORIGIN
1 gaccaccccc accgccctca aagtagacgg catcgcagct tggatacacg ccgcccacgt
61 gaaggctgcc gaccccgggg gtggaccatc ctctagactg ccggatctag ctagttaatt
121 aaggatccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
181 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
241 ccacctacgg caagctgacc ctgaagctga tctgcaccac cggcaagctg cccgtgccct
301 ggcccaccct cgtgaccacc ctgggctacg gcctgcagtg cttcgcccgc taccccgacc
361 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
421 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
481 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
541 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcacc gccgacaagc
601 agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc
661 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
721 acaaccacta cctgagctac cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
781 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
841 acaagggata tccatcacac tggcggccgg aattgccaaa aaagaagaga aaggtagaat
901 tcatggatat agaaaatgag cagacactga atgtgaaccc cactgaccct gacaatttaa
961 gcgactctct cttttctgga gatgaagaaa atgctggcac tgaagaaata aagaatgaaa
1021 taaatggaaa ttggatttcc gcatctacta ttaatgaagc tagaatcaat gcaaaagcca
1081 aaagacgact gcggaaaaat tcatcccggg actctggccg aggagactca gtcagtgaca
1141 attgagcggc cgctacgtaa attccgcccc ccccccccct ctccctcccc cccccctaac
1201 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
1261 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
1321 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
1381 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
1441 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
1501 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
1561 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
1621 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg
1681 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
1741 cgatgataat atggccacaa ccatggttac cgagtacaag cccacggtgc gcctcgccac
1801 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
1861 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
1921 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
1981 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
2041 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
2101 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
2161 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
2221 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
2281 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
2341 catgacccgc aagcccggtg cctgagtcga cgataaaata aaagatttta tttagtctcc
2401 agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc
2461 cattttgcaa ggcatggaaa aatacataac tgagaataga gaagttcaga tcaaggtcag
2521 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
2581 ccggctcagg gccaagaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg
2641 taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtccagcc
2701 ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac
2761 cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg
2821 ctccccgagc tcaataaaag agcccacaac ccctcactcg gggcgccagt cctccgattg
2881 actgagtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt
2941 ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc
3001 tttcacatgc agcatgtatc aaaattaatt tggttttttt tcttaagtat ttacattaaa
3061 tggccatagt tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
3121 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
3181 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
3241 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
3301 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
3361 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
3421 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
3481 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
3541 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
3601 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
3661 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
3721 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
3781 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
3841 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
3901 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
3961 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttgcggcc ggccgcaaat
4021 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
4081 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
4141 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
4201 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
4261 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
4321 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
4381 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
4441 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
4501 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
4561 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
4621 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
4681 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
4741 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
4801 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
4861 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
4921 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
4981 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
5041 tgccacctgc ataatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc
5101 attttgcaag gcatggaaaa atacataact gagaatagaa aagttcagat caaggtcagg
5161 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
5221 cggctcaggg ccaagaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt
5281 aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc ggtccagccc
5341 tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc
5401 ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc
5461 tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc ctccgattga
5521 ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg
5581 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct
5641 ttcatttggg ggctcgtccg ggatcgggag acccctgccc agggaccacc gacccaccac
5701 cgggaggtaa gctggccagc aacttatctg tgtctgtccg attgtctagt gtctatgact
5761 gattttatgc gcctgcgtcg gtactagtta gctaactagc tctgtatctg gcggacccgt
5821 ggtggaactg acgagttcgg aacacccggc cgcaaccctg ggagacgtcc cagggacttc
5881 gggggccgtt tttgtggccc gacctgagtc caaaaatccc gatcgttttg gactctttgg
5941 tgcacccccc taataggagg gatatgtggt tctggtagga gacgagaacc taaaacagtt
6001 cccgcctccg tctgaatttt tgctttcggt ttgggaccga agccgcgccg cgcgtcttgt
6061 ctgctgcagc atcgttctgt gttgtctctg tctgactgtg tttctgtatt tgtctgaaaa
6121 ttagggccag actgttacca ctcccttaag tttgacctta ggtcactgga aagatgtcga
6181 gcggatcgct cacaaccagt cggtagatgt caagaagaga cgttgggtta ccttctgctc
6241 tgcagaatgg ccaaccttta acgtcggatg gccgcgagac ggcaccttta accgagacct
6301 catcacccag gttaagatca aggtcttttc acctggcccg catggacacc cagaccaggt
6361 cccctacatc gtgacctggg aagccttggc ttttgacccc cctccctggg tcaagccctt
6421 tgtacaccct aagcctccgc ctcctcttcc tccatccgcc ccgtctctcc cccttgaacc
6481 tcctcgttcg accccgcctc gatcctccct ttatccagcc ctcactcctt ctctaggcgc
6541 ccccatatgg ccatatgaga tcttatatgg ggcacccccg ccccttgtaa acttccctga
6601 ccctgacatg acaagagtta ctaacagccc ctctctccaa gctcacttac aggctctcta
6661 cttagtccag cacgaagtct ggagacctct ggcggcagcc taccaagaac aactggaccg
6721 accggtggta cctcaccctt accgagtcgg cgacacagtg tgggtccgcc gacaccagac
6781 taagaaccta gaacctcgct ggaaaggacc ttacacagtc ctgct
//
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