| 质粒名称: | pMSCVpuro-mito-Pericam |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 87381 |
| 存储实验室: | Björn Stork |
|
载体骨架基本信息 |
|
| 载体名称: | pMSCVpuro |
| 载体出品公司: | Clontech |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6264 bp |
| 完整质粒大小: | 7740 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 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Mito-Pericam [ratiometric] |
| 物种来源: | -- |
| 插入大小: | 1488 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | Retroviral 5' LTR promoter |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | NheI(未破坏) |
| 3' 克隆位点: | EcoRI(未破坏) |
| 5' 测序引物 | pMSCV-F |
| 3' 测序引物 | pMSCV-R |
|
文献引用方法 |
|
| 材料和方法部分: | pMSCVpuro-Mito-Pericam was a gift from Björn Stork (Addgene plasmid # 87381 ; http://n2t.net/addgene:87381 ; RRID:Addgene_87381) |
| 参考文献部分: | The mycotoxin phomoxanthone A disturbs the form and function of the inner mitochondrial membrane. Bohler P, Stuhldreier F, Anand R, Kondadi AK, Schlutermann D, Berleth N, Deitersen J, Wallot-Hieke N, Wu W, Frank M, Niemann H, Wesbuer E, Barbian A, Luyten T, Parys JB, Weidtkamp-Peters S, Borchardt A, Reichert AS, Pena-Blanco A, Garcia-Saez AJ, Itskanov S, van der Bliek AM, Proksch P, Wesselborg S, Stork B. Cell Death Dis. 2018 Feb 19;9(3):286. doi: 10.1038/s41419-018-0312-8. 10.1038/s41419-018-0312-8 PubMed 29459714 |
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 pMSCVpuro-Mito-Pericam 7740 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Retroviral transfection of cells to express ratiometric Pericam as
described by Nagai et al 2001 (PMID 11248055) localising to the
mitochondrial matrix via the transit peptide from human COX8A..
ACCESSION .
VERSION .
KEYWORDS pMSCVpuro-Mito-Pericam
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7740)
AUTHORS Bohler P, Stuhldreier F, Anand R, Kondadi AK, Schlutermann D,
Berleth N, Deitersen J, Wallot-Hieke N, Wu W, Frank M, Niemann H,
Wesbuer E, Barbian A, Luyten T, Parys JB, Weidtkamp-Peters S,
Borchardt A, Reichert AS, Pena-Blanco A, Garcia-Saez AJ, Itskanov S,
van der Bliek AM, Proksch P, Wesselborg S, Stork B
TITLE The mycotoxin phomoxanthone A disturbs the form and function of the
inner mitochondrial membrane.
JOURNAL Cell Death Dis. 2018 Feb 19;9(3):286. doi:
10.1038/s41419-018-0312-8.
PUBMED 29459714
REFERENCE 2 (bases 1 to 7740)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7740
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 1..517
/label=5' LTR
/note="5' long terminal repeat from murine embryonic stem
cell virus"
misc_feature 581..922
/label=MESV Psi
/note="packaging signal of murine embryonic stem cell
virus"
CDS 989..1405
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 1335..1357
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 1373..1389
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
CDS 1625..1702
/codon_start=1
/product="calmodulin-binding peptide"
/label=CBP
/note="derived from skeletal muscle myosin light chain
kinase; binds calmodulin with nanomolar affinity in the
presence of calcium"
/translation="KRRWKKNFIAVSAANRFKKISSSGAL"
primer_bind 1933..1954
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(2066..2087)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2327..2346)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
promoter 2916..3415
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
primer_bind complement(2951..2974)
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
primer_bind 3348..3367
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
CDS 3436..4035
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(3436..3455)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 3932..3952
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
LTR 4135..4649
/label=3' LTR
/note="3' long terminal repeat from murine embryonic stem
cell virus"
protein_bind 4811..4827
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(4835..4865)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4880..4901
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5018..5035)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5189..5777)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5269..5288)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5948..6808)
/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 6571..6590
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6809..6913)
/gene="bla"
/label=AmpR promoter
primer_bind 6981..6999
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(7037..7059)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 7159..7178
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 7372..7394
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 7522..7541
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
ORIGIN
1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
1381 agccctcact ccttctctag gcgccggaat tagatccgct agcatgtccg tcctgacgcc
1441 gctgctgctg cggggcttga caggctcggc ccggcggctc ccagtgccgc gcgccaagat
1501 ccattcgttg ggggacccca tgtccgtcct gacgccgctg ctgctgcggg gcttgacagg
1561 ctcggcccgg cggctcccag tgccgcgcgc caagatccat tcgttgccgc cggagggaga
1621 tctcaagagg cgctggaaga aaaacttcat tgccgtcagc gctgccaacc ggttcaagaa
1681 gatctccagc tccggggcac tggggtctgc aggctacaac agcgacaacg tctatatcat
1741 ggccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga
1801 cggcggcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
1861 gctgctgccc gacaaccact acctgagctt ccagtccgcc ctgagcaaag accccaacga
1921 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
1981 ggacgagctg tacaaggtcg acggtggcag cggtggcacc ggtgtgagca agggcgagga
2041 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
2101 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct
2161 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttcggcta
2221 cggcctgaag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
2281 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
2341 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
2401 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaacggtac
2461 cgaccaactg acagaagagc agattgcaga gttcaaagaa gccttctcat tattcgacaa
2521 ggatggggac ggcaccatca ccacaaagga acttggcacc gttatgaggt cgcttggaca
2581 aaacccaacg gaagcagaat tgcaggatat gatcaatgaa gtcgatgctg atggcaatgg
2641 aacgatttac tttcctgaat ttcttactat gatggctaga aaaatgaagg acacagacag
2701 cgaagaggaa atccgagaag cattccgtgt ttttgacaag gatgggaacg gctacatcag
2761 cgctgctcag ttacgtcacg tcatgacaaa cctcggggag aagttaacag atgaagaagt
2821 tgatgaaatg ataagggaag cagatatcga tggtgatggc caagtaaact atgaagagtt
2881 tgtacaaatg atgacagcaa agtaagaatt ctaccgggta ggggaggcgc ttttcccaag
2941 gcagtctgga gcatgcgctt tagcagcccc gctgggcact tggcgctaca caagtggcct
3001 ctggcctcgc acacattcca catccaccgg taggcgccaa ccggctccgt tctttggtgg
3061 ccccttcgcg ccaccttcta ctcctcccct agtcaggaag ttcccccccg ccccgcagct
3121 cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc actagtctcg tgcagatgga
3181 cagcaccgct gagcaatgga agcgggtagg cctttggggc agcggccaat agcagctttg
3241 ctccttcgct ttctgggctc agaggctggg aaggggtggg tccgggggcg ggctcagggg
3301 cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat tctgcacgct
3361 tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca tctccgggcc tttcgacctg
3421 cagcccaagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
3481 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
3541 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
3601 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
3661 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
3721 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
3781 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
3841 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
3901 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
3961 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
4021 aagcccggtg cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga
4081 ccccatgcat cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa
4141 gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa
4201 atacataact gagaatagag aagttcagat caaggttagg aacagagaga cagcagaata
4261 tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga
4321 tggtccccag atgcggtccc gccctcagca gtttctagag aaccatcaga tgtttccagg
4381 gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc
4441 tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac
4501 tcggcgcgcc agtcctccga tagactgcgt cgcccgggta cccgtgtatc caataaaccc
4561 tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga
4621 ttgactaccc gtcagcgggg gtctttcatg ggtaacagtt tcttgaagtt ggagaacaac
4681 attctgaggg taggagtcga atattaagta atcctgactc aattagccac tgttttgaat
4741 ccacatactc caatactcct gaaatagttc attatggaca gcgcagaaag agctggggag
4801 aattgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
4861 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
4921 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
4981 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
5041 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
5101 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
5161 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
5221 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
5281 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
5341 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
5401 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
5461 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
5521 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
5581 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
5641 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
5701 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
5761 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
5821 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
5881 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
5941 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
6001 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
6061 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
6121 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
6181 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
6241 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
6301 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
6361 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
6421 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
6481 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
6541 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
6601 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
6661 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
6721 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
6781 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
6841 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
6901 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
6961 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
7021 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
7081 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
7141 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
7201 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca
7261 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
7321 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
7381 gacgttgtaa aacgacggcg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc
7441 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
7501 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
7561 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg cgattagtcc aatttgttaa
7621 agacaggata tcagtggtcc aggctctagt tttgactcaa caatatcacc agctgaagcc
7681 tatagagtac gagccataga taaaataaaa gattttattt agtctccaga aaaagggggg
//
