| 质粒名称: | 22l-COX8-3xFlag-TALE(RNR2-Right)-TadA(8e+V106W) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 209296 |
| 存储实验室: | Vladislav Verkhusha |
|
载体骨架基本信息 |
|
| 载体名称: | pCMV |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 6040bp |
| 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: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | eCOX8-3xFlag-TALE(RNR2-Right)-TadA(8e+V106W) |
| 物种来源: | H. sapiens (human) |
|
文献引用方法 |
|
| 材料和方法部分: | 22l-COX8-3xFlag-TALE(RNR2-Right)-TadA(8e+V106W) was a gift from Wensheng Wei (Addgene plasmid # 209296 ; http://n2t.net/addgene:209296 ; RRID:Addgene_209296) |
| 参考文献部分: | Strand-selective base editing of human mitochondrial DNA using mitoBEs. Yi Z, Zhang X, Tang W, Yu Y, Wei X, Zhang X, Wei W. Nat Biotechnol. 2023 May 22. doi: 10.1038/s41587-023-01791-y. 10.1038/s41587-023-01791-y PubMed 37217751 |
买家导航
22l-COX8-3xFlag-TALE(RNR2-Right)-TadA(8e+V106W)载体质粒基本信息
22l-COX8-3xFlag-TALE(RNR2-Right)-TadA(8e+V106W)质粒图谱载体图谱和 22l-COX8-3xFlag-TALE(RNR2-Right)-TadA(8e+V106W)载体序列质粒序列多克隆位点信息
22l-COX8-3xFlag-TALE(RNR2-Right)-TadA(8e+V106W)质粒载体简介
22l-COX8-3xFlag-TALE(RNR2-Right)-TadA(8e+V106W)质粒序列载体序列
LOCUS 22l-COX8-3xFlag-TALE(RNR2-Right)-TadA(8e+V106W) 6040 bp DNA circular SYN 14-APR-2025
DEFINITION mitoBE expression vector, target mtRNR2 gene..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6040)
AUTHORS Yi Z, Zhang X, Tang W, Yu Y, Wei X, Zhang X, Wei W
TITLE Strand-selective base editing of human mitochondrial DNA using
mitoBEs.
JOURNAL Nat Biotechnol. 2023 May 22. doi: 10.1038/s41587-023-01791-y.
PUBMED 37217751
REFERENCE 2 (bases 1 to 6040)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Biotechnol. 2023 May 22. doi: 10.1038/s41587-023-01791-y."
COMMENT Sequence Label: 22l-COX8-3xFlag-TALE(RNR2-Right)-TadA(8e+V106W)
FEATURES Location/Qualifiers
source 1..6040
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 132..335
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 286..306
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 377..396
/label=T7
/note="T7 promoter, forward primer"
promoter 377..395
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 421..430
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 427..504
/codon_start=1
/product="mitochondrial presequence of human cytochrome c
oxidase subunit VIII (Rizutto et al., 1989)"
/label=COX8 presequence
/translation="MASVLTPLLLRGLTGSARRLPVPRAK"
CDS 517..582
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
primer_bind complement(3396..3412)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3396..3412)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3409..3431)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3420..3436
/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(join(3444..3450,3451..3468,3469..3474))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3489..3510
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3627..3644)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3798..4386)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3878..3897)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(4557..5348,5349..5417))
/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 5180..5199
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5418..5522)
/gene="bla"
/label=AmpR promoter
primer_bind complement(5601..5620)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer join(5792..6040,1..131)
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
ORIGIN
1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
361 agagatccgc ggccgctaat acgactcact atagggagac ccaagctggc tagcaccgcc
421 gccaccatgg cctccgttct gaccccgctg ctgctgcgtg gcctgaccgg ctccgctcgt
481 cgtctgccag ttccgcgtgc gaaaatccat tccctggact acaaagacca tgacggtgat
541 tataaagatc atgacatcga ttacaaggat gacgatgaca agatggacat cgcggacctc
601 aggacgctgg gctacagcca gcaacagcaa gagaagatca agcccaaagt aaggagcacc
661 gtggcccagc accacgaggc cctggtgggt cacggcttca cccacgcgca tatcgttgct
721 ctgagccaac atcccgcagc tctgggtacc gttgcggtga agtatcagga catgatcgcg
781 gcactgcctg aagctacaca cgaagccata gtgggcgttg gcaagcagtg gagcggtgcc
841 agagcgcttg aggcactgtt gacggtggct ggcgagctga ggggaccgcc actgcaactg
901 gacaccggcc aactgctgaa gatcgccaag aggggaggcg tgacggcggt ggaggccgtg
961 catgcctgga ggaacgcact gaccggcgcg cccctgaacc tgacaccaga gcaagtagtg
1021 gctattgcaa gtaacaacgg tggcaaacaa gcgctggaga ccgtgcagag gctccttccg
1081 gtgctctgcc aagcacacgg tctcactccg gaacaggtgg tcgcaatcgc gagccacgac
1141 ggcggcaagc aagcccttga gacagtccaa agacttttgc ctgtcctttg tcaggcgcat
1201 ggccttacgc ctgagcaagt cgttgcgatc gcctccaaca tcggcggaaa acaggctttg
1261 gaaaccgtgc agcggttgct gcccgttttg tgccaagccc acggattgac ccccgaacag
1321 gttgtagcca tagcttctca cgacggaggt aagcaggcac tggaaaccgt gcagcgcctg
1381 ctcccagtac tgtgtcaggc tcatgggctg acaccagagc aagtagtggc tattgcaagt
1441 cacgacggtg gcaaacaagc gctggagacc gtgcagaggc tccttccggt gctctgccaa
1501 gcacacggtc tcactccgga acaggtggtc gcaatcgcga gcaacatcgg cggcaagcaa
1561 gcccttgaga cagtccaaag acttttgcct gtcctttgtc aggcgcatgg ccttacgcct
1621 gagcaagtcg ttgcgatcgc ctccaacggc ggcggaaaac aggctttgga aaccgtgcag
1681 cggttgctgc ccgttttgtg ccaagcccac ggattgaccc ccgaacaggt tgtagccata
1741 gcttctcacg acggaggtaa gcaggcactg gaaaccgtgc agcgcctgct cccagtactg
1801 tgtcaggctc atgggctcac tccggaacag gtggtcgcaa tcgcgagcaa caacggcggc
1861 aagcaagccc ttgagacagt ccaaagactt ttgcctgtcc tttgtcaggc gcatggcctt
1921 acgcctgagc aagtcgttgc gatcgcctcc aacaacggcg gaaaacaggc tttggaaacc
1981 gtgcagcggt tgctgcccgt tttgtgccaa gcccacggat tgacccccga acaggttgta
2041 gccatagctt ctaacaacgg aggtaagcag gcactggaaa ccgtgcagcg cctgctccca
2101 gtactgtgtc aggctcatgg gctgacacca gagcaagtag tggctattgc aagtaacatc
2161 ggtggcaaac aagcgctgga gaccgtgcag aggctccttc cggtgctctg ccaagcacac
2221 ggtctcactc cggaacaggt ggtcgcaatc gcgagcaacg gcggcggcaa gcaagccctt
2281 gagacagtcc aaagactttt gcctgtcctt tgtcaggcgc atggccttac gcctgagcaa
2341 gtcgttgcga tcgcctccaa caacggcgga aaacaggctt tggaaaccgt gcagcggttg
2401 ctgcccgttt tgtgccaagc ccacggattg acccccgaac aggttgtagc catagcttct
2461 aacggcggag gtaagcaggc actggaaagc atcgtggccc agctgagcag acccgaccct
2521 gccctcgccg ctctgaccaa cgaccacctg gtggccctgg cttgcctcgg gggcaggcca
2581 gctcttgacg ccgtgaagaa gggccttggt ggatcctctg aggtggagtt ttcccacgag
2641 tactggatga gacatgccct gaccctggcc aagagggcac gcgatgagag ggaggtgcct
2701 gtgggagccg tgctggtgct gaacaataga gtgatcggcg agggctggaa cagagccatc
2761 ggcctgcacg acccaacagc ccatgccgaa attatggccc tgagacaggg cggcctggtc
2821 atgcagaact acagactgat tgacgccacc ctgtacgtga cattcgagcc ttgcgtgatg
2881 tgcgccggcg ccatgatcca ctctaggatc ggccgcgtgg tgtttggctg gaggaactcc
2941 aaaaggggcg ccgcaggctc cctgatgaac gtgctgaatt accccggcat gaatcaccgc
3001 gtcgaaatta ccgagggaat cctggcagat gaatgtgccg ccctgctgtg cgatttctac
3061 cggatgccta gacaggtgtt caatgctcag aagaaggccc agagctccat caattgataa
3121 ggggtctttg tcctctgtac tgtctctctc cttgccccta acccaaaaag cttcattttt
3181 ctgtgtaggc tgcacaagag ccttgattga agatatattc tttctgaaca gtatttaagg
3241 tttccaataa aatgtacacc cctcagaaac cggtgtttaa acgaagacaa tagcaggcat
3301 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctcgata
3361 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
3421 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctag
3481 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
3541 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
3601 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
3661 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
3721 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
3781 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
3841 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
3901 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
3961 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
4021 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
4081 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
4141 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
4201 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
4261 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
4321 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
4381 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
4441 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
4501 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
4561 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
4621 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
4681 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
4741 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
4801 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
4861 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
4921 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
4981 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
5041 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
5101 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
5161 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
5221 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
5281 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
5341 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
5401 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
5461 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
5521 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatcg atctcccgat
5581 cccctagggt cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
5641 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
5701 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
5761 cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag
5821 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
5881 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
5941 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
6001 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
//
