质粒名称: | Snap-Omp25 |
---|---|
出品公司: | Addgene |
目录编号: | 69599 |
存储实验室: | David Sabatini |
载体骨架基本信息 |
|
载体名称: | pLJM2 |
载体出品公司: | Jason Moffat |
空载体大小: | -- |
完整质粒大小: | 8017 bp |
载体修饰: | -- |
载体类型: | 哺乳细胞表达 |
筛选标记: | 嘌呤霉素 |
载体抗性: | 氨苄霉素 |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | -- |
插入基因信息 |
|
插入基因名称: | Snap-Omp25 |
物种来源: | -- |
插入大小: | -- |
基因突变: | -- |
GenBank编号 | -- |
启动子: | -- |
标签/融合蛋白: | -- |
克隆方法: | 限制性内切酶 |
5’克隆位点: | Nhe1(未破坏) |
3’克隆位点: | EcoR1(未破坏) |
5’测序引物 | CMV-F |
3’测序引物 | -- |
文献引用方法 |
|
材料和方法部分: | Snap-Omp25 was a gift from David Sabatini (Addgene plasmid # 69599 ; http://n2t.net/addgene:69599 ; RRID:Addgene_69599) |
参考文献部分: | Stem cells. Asymmetric apportioning of aged mitochondria between daughter cells is required for stemness. Katajisto P, Dohla J, Chaffer CL, Pentinmikko N, Marjanovic N, Iqbal S, Zoncu R, Chen W, Weinberg RA, Sabatini DM. Science. 2015 Apr 17;348(6232):340-3. doi: 10.1126/science.1260384. Epub 2015 Apr 2. 10.1126/science.1260384 PubMed 25837514 |
买家导航
SnapOmp25载体基本信息
SnapOmp25载体图谱质粒图谱和多克隆位点信息
SnapOmp25载体简介
SnapOmp25载体序列
LOCUS Snap-Omp25 8017 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Snap-tag substrates with two different fluorophores (red and green) sequentially to separately label young and old mitochondria. ACCESSION . VERSION . KEYWORDS Snap-Omp25 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8017) AUTHORS Katajisto P, Dohla J, Chaffer CL, Pentinmikko N, Marjanovic N, Iqbal S, Zoncu R, Chen W, Weinberg RA, Sabatini DM TITLE Stem cells. Asymmetric apportioning of aged mitochondria between daughter cells is required for stemness. JOURNAL Science. 2015 Apr 17;348(6232):340-3. doi: 10.1126/science.1260384. Epub 2015 Apr 2. PUBMED 25837514 REFERENCE 2 (bases 1 to 8017) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8017 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 131..334 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 285..305 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 366..911 /codon_start=1 /product="human O6-alkylguanine-DNA-alkyltransferase variant that forms covalent bonds with benzylguanine derivatives" /label=SNAP-tag(R) /note="optimized for mammalian expression" /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHEIKLLGKGTSAADAVE VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK VVKFGEVISYQQLAALAGNPAATAAVKTALSGNPVPILIPCHRVVSSSGAVGGYEGGLA VKEWLLAHEGHRLGKPGLG" misc_feature 1100..1217 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 1266..1776 /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" primer_bind 1695..1713 /label=hPGK-F /note="Human PGK promoter, forward primer" CDS 1798..2397 /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(1798..1817) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 2294..2314 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 2525..2758 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 2830..2951 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2867..2886) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2921..2940 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 2991..3126 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3053..3072 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(3146..3165) /label=T7 /note="T7 promoter, forward primer" promoter complement(3147..3165) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3175..3192) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(3175..3191) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3184..3206) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 3333..3788 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3420..3439) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3630..3651 /label=F1ori-F /note="F1 origin, forward primer" promoter 3814..3918 /gene="bla" /label=AmpR promoter CDS 3919..4779 /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 complement(4137..4156) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4950..5538 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5439..5458 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5692..5709 /label=L4440 /note="L4440 vector, forward primer" protein_bind 5826..5847 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5862..5892 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5900..5916 /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)." primer_bind 5905..5927 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 5924..5940 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5924..5940 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 5959..5979 /label=T3 /note="T3 promoter, forward primer" promoter 5961..5979 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 6007..6233 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 6234..6414 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 6461..6586 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 7079..7312 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 7497..7541 /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" primer_bind 7839..7859 /label=hU6-F /note="Human U6 promoter, forward primer" ORIGIN 1 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc 61 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc 121 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc 181 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa 241 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag 301 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta 361 gcgccatgga caaagattgc gaaatgaaac gtaccaccct ggatagccct ctgggcaagc 421 tggaactgtc tgggtgcgaa cagggcctgc acgagatcaa gctgctgggc aaaggaacat 481 ctgccgccga cgccgtggaa gtgcctgccc cagccgccgt gctgggcgga ccagagccac 541 tgatgcaggc caccgcctgg ctcaacgcct actttcacca gcctgaggcc atcgaggagt 601 tccctgtgcc agccctgcac cacccagtgt tccagcagga gagctttacc cgccaggtgc 661 tgtggaaact gctgaaagtg gtgaagttcg gagaggtcat cagctaccag cagctggccg 721 ccctggccgg caatcccgcc gccaccgccg ccgtgaaaac cgccctgagc ggaaatcccg 781 tgcccattct gatcccctgc caccgggtgg tgtctagctc tggcgccgtg gggggctacg 841 agggcgggct cgccgtgaaa gagtggctgc tggcccacga gggccacaga ctgggcaagc 901 ctgggctggg tcctgcaggt ataggcgcgc caggatccac gcgtgggcga ggcgacggag 961 agccgagtgg agttcctgta gctgtggtgc tgctgccagt gtttgccctt accctggtag 1021 cagtttgggc cttcgtgaga taccgaaagc agctctgaga attctcgacc tcgagacaaa 1081 tggcagtatt catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg 1141 aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta 1201 caaaaattca aaattttcgg gtttattaca gggacagcag agatccactt tggccgcggc 1261 tcgagggggt tggggttgcg ccttttccaa ggcagccctg ggtttgcgca gggacgcggc 1321 tgctctgggc gtggttccgg gaaacgcagc ggcgccgacc ctgggtctcg cacattcttc 1381 acgtccgttc gcagcgtcac ccggatcttc gccgctaccc ttgtgggccc cccggcgacg 1441 cttcctgctc cgcccctaag tcgggaaggt tccttgcggt tcgcggcgtg ccggacgtga 1501 caaacggaag ccgcacgtct cactagtacc ctcgcagacg gacagcgcca gggagcaatg 1561 gcagcgcgcc gaccgcgatg ggctgtggcc aatagcggct gctcagcagg gcgcgccgag 1621 agcagcggcc gggaaggggc ggtgcgggag gcggggtgtg gggcggtagt gtgggccctg 1681 ttcctgcccg cgcggtgttc cgcattctgc aagcctccgg agcgcacgtc ggcagtcggc 1741 tccctcgttg accgaatcac cgacctctct ccccaggggg atccaccgga gcttaccatg 1801 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc 1861 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc 1921 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc 1981 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc 2041 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc 2101 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc 2161 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc 2221 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag 2281 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac 2341 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacgc 2401 ccgccccacg acccgcagcg cccgaccgaa aggagcgcac gaccccatgc atcggtacct 2461 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg 2521 ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg 2581 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg 2641 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt 2701 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt 2761 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag 2821 agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 2881 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 2941 aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc atcccgcccc 3001 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg 3061 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg 3121 gaggcctagg gacgtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc 3181 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc 3241 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc 3301 ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc 3361 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc 3421 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct 3481 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa 3541 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc 3601 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact 3661 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg 3721 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct 3781 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc 3841 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa 3901 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt 3961 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct 4021 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc 4081 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta 4141 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac 4201 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc 4261 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac 4321 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg 4381 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac 4441 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc 4501 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt 4561 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga 4621 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc 4681 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag 4741 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca 4801 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc 4861 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca 4921 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc 4981 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta 5041 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt 5101 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc 5161 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg 5221 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg 5281 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag 5341 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc 5401 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat 5461 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg 5521 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc 5581 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt 5641 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca 5701 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg 5761 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac 5821 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg 5881 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac 5941 catgattacg ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctgcaa 6001 gcttaatgta gtcttatgca atactcttgt agtcttgcaa catggtaacg atgagttagc 6061 aacatgcctt acaaggagag aaaaagcacc gtgcatgccg attggtggaa gtaaggtggt 6121 acgatcgtgc cttattagga aggcaacaga cgggtctgac atggattgga cgaaccactg 6181 aattgccgca ttgcagagat attgtattta agtgcctagc tcgatacata aacgggtctc 6241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 6301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 6361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg 6421 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg 6481 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat 6541 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg 6601 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa 6661 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct 6721 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac 6781 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca 6841 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa 6901 aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga 6961 gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag 7021 tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag 7081 gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga 7141 cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc 7201 tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc 7261 tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt 7321 ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta 7381 ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta 7441 acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga 7501 atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa 7561 caaattggct gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa 7621 gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat 7681 cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag 7741 aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatct cgacggtatc 7801 gatcacgaga ctagcctcga gcggccgccc ccttcaccga gggcctattt cccatgattc 7861 cttcatattt gcatatacga tacaaggctg ttagagagat aattggaatt aatttgactg 7921 taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt 7981 ttgcagtttt aaaattatgt tttaaaatgg actatca //
SnapOmp25载体图谱质粒图谱pdf版和相关资料下载
SnapOmp25载体质粒应用举例
- 上一篇:pMK284 (Stag-3FLAG-Hygro)
- 下一篇:CMV-SEAP