| 质粒名称: | pDF0159 pCMV - huDisCas7-11 mammalian expression |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 172507 |
| 存储实验室: | Omar Abudayyeh, Jonathan Gootenberg |
|
载体骨架基本信息 |
|
| 载体名称: | Unspecified |
| 载体出品公司: | -- |
| 完整质粒大小: | 8205 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄青霉素，100μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称: | pCMV-huDisCas7-11哺乳动物表达 |
| 物种来源: | 石本脱硫虫 |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | Gibson Cloning |
|
文献引用方法 |
|
| 材料和方法部分: | pDF0159 pCMV - huDisCas7-11 mammalian expression was a gift from Omar Abudayyeh & Jonathan Gootenberg (Addgene plasmid # 172507 ; http://n2t.net/addgene:172507 ; RRID:Addgene_172507) |
| 参考文献部分: | Programmable RNA targeting with the single-protein CRISPR effector Cas7-11. Ozcan A, Krajeski R, Ioannidi E, Lee B, Gardner A, Makarova KS, Koonin EV, Abudayyeh OO, Gootenberg JS. Nature. 2021 Sep 6. pii: 10.1038/s41586-021-03886-5. doi: 10.1038/s41586-021-03886-5. 10.1038/s41586-021-03886-5 PubMed 34489594 |
买家导航
pDF0159 pCMV - huDisCas7-11 mammalian expression载体质粒基本信息
pDF0159 pCMV - huDisCas7-11 mammalian expression质粒图谱载体图谱和pDF0159 pCMV - huDisCas7-11 mammalian expression载体序列质粒序列多克隆位点信息
pDF0159 pCMV - huDisCas7-11 mammalian expression质粒载体简介
pDF0159 pCMV - huDisCas7-11 mammalian expression质粒序列载体序列
LOCUS pDF0159 pCMV - huDisCas7-11 mammalian expression 8205 bp ds-DNA circular SYN 07-OCT-2021
DEFINITION Encodes human codon optimized DisCas7-11 protein alone for
expression in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pDF0159 pCMV - huDisCas7-11 mammalian expression
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8205)
AUTHORS Ozcan A, Krajeski R, Ioannidi E, Lee B, Gardner A, Makarova KS,
Koonin EV, Abudayyeh OO, Gootenberg JS
TITLE Programmable RNA targeting with the single-protein CRISPR effector
Cas7-11.
JOURNAL Nature. 2021 Sep 6. pii: 10.1038/s41586-021-03886-5. doi:
10.1038/s41586-021-03886-5.
PUBMED 34489594
REFERENCE 2 (bases 1 to 8205)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Oct 7, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8205
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(70..89)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 261..640
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 641..844
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 795..815
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 886..905
/label=T7
/note="T7 promoter, forward primer"
promoter 886..904
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 5729..5746
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(5769..5786)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5775..5999
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(6070..6086)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6070..6086)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6083..6105)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6094..6110
/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(6118..6148)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6163..6184
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6301..6318)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6472..7060)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6552..6571)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7231..8091)
/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 7854..7873
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8092..8196)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ccgaaaagtg ccacctgacg tcgacggatc gggagatcga tctcccgatc ccctagggtc
61 tactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
121 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
181 gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt
241 acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac
301 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
361 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
421 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
481 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
541 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
601 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
661 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
721 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
781 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
841 agctggttta gtgaaccgtc agatccgcta gagatccgcg gccgctaata cgactcacta
901 tagggagagc cgccaccatg acgactacta tgaagatttc aattgaattc ctcgagccgt
961 ttcggatgac caaatggcag gaaagcacta ggcgaaacaa gaacaacaag gaatttgttc
1021 gcggccaagc ttttgcaaga tggcatagaa ataagaagga taacacaaaa gggcggcctt
1081 acataactgg gacattgctg cggtccgcag taattagaag cgcggaaaac ctgctgacat
1141 tgagtgatgg gaaaattagc gaaaagacat gttgtcctgg gaaatttgat acggaagata
1201 aagacaggtt gctgcaattg aggcagcggt ccacccttcg ttggaccgat aagaacccct
1261 gtccagataa cgctgaaact tactgcccgt tttgcgaact tctcgggaga tccggtaacg
1321 atggaaagaa ggctgagaag aaggattggc gctttcgtat tcattttggc aatctttccc
1381 ttccagggaa gcccgatttc gacgggccaa aagctatagg cagccaacgg gtacttaaca
1441 gggtcgattt caaatccggt aaggcccatg acttctttaa ggcttacgaa gtcgaccata
1501 ctaggtttcc ccgcttcgaa ggggagatta ccatagataa taaagtcagc gctgaagcca
1561 ggaaactgct ctgtgattct ctcaagttta cggatcggct gtgtggagct ctgtgcgtaa
1621 tcaggttcga tgaatataca ccagcagccg atagtgggaa acaaaccgaa aatgtccaag
1681 cagaaccgaa tgctaatctc gctgaaaaga ccgcagagca aattattagc attctggacg
1741 ataataagaa aaccgaatat acccgcctgc ttgctgatgc tattcgttca ctccgccgct
1801 ctagtaaact tgtcgctggc ttgccaaagg atcatgacgg gaaggatgac cattacctct
1861 gggatatcgg caagaagaag aaagacgaaa actctgtcac tattaggcaa atccttacga
1921 cctcagcaga caccaaggaa ctcaagaatg ccggaaaatg gagagaattc tgcgagaagc
1981 tgggtgaagc gctgtacctc aagagtaaag acatgagtgg cggcctgaaa attactcgga
2041 gaatactggg cgatgctgaa ttccatggaa agcccgatcg gcttgaaaag agccggtctg
2101 tgtcaatcgg atctgtgttg aaagaaactg tggtatgcgg cgaactggtc gctaaaacgc
2161 ccttcttctt cggagcgata gacgaagatg caaaacaaac cgacctgcaa gtactcctca
2221 ctcccgataa caagtataga ctgccaagaa gcgccgtgcg aggtatactc cgtcgagatc
2281 ttcaaaccta ttttgatagc ccatgcaatg ctgagttggg tggacggcca tgcatgtgta
2341 agacgtgtag gattatgaga gggatcacgg tgatggatgc gcgcagtgag tacaatgccc
2401 cgccagaaat aaggcatcgt acccgcatta atcccttcac aggcacggtt gcggaaggtg
2461 ccctgtttaa tatggaagta gcccccgagg ggattgtctt tccatttcaa ctccggtacc
2521 ggggctctga agatgggctg cccgatgcac tgaaaacggt gttgaaatgg tgggctgagg
2581 ggcaggcatt catgagtggc gctgcctcaa ccgggaaggg ccgattccgg atggaaaatg
2641 ctaaatatga aacgctggat ctgagcgacg agaatcaaag gaatgactat cttaagaatt
2701 ggggatggcg tgacgagaag gggctcgagg aactgaagaa acgactgaac tcaggtctgc
2761 cagagcccgg taattatagg gatccaaaat ggcacgagat taacgtttcc attgagatgg
2821 caagcccttt tattaatggc gacccaatcc gcgcagccgt ggacaaacgt ggtacagatg
2881 tggttacctt cgttaagtat aaagctgaag gggaagaggc gaaacccgta tgtgcataca
2941 aggccgaatc ttttagaggg gtgatcagaa gtgccgtggc acgcattcat atggaagatg
3001 gcgtcccttt gactgagttg actcacagtg actgtgaatg tctcctgtgc caaatctttg
3061 gaagtgagta tgaagccggc aaaataaggt ttgaagatct cgtattcgaa agtgacccgg
3121 aacctgtgac cttcgatcat gtggccatcg atagattcac tggcggtgca gctgataaga
3181 agaaattcga tgattcccct ctgcccggta gccctgcaag accgctcatg ttgaaaggct
3241 ccttctggat ccgcagggac gttctcgagg acgaagagta ctgtaaggca ctcggtaagg
3301 ctcttgcaga tgtgaataat ggcctttatc ccctcggtgg aaagagcgcc atcggctacg
3361 gacaggtcaa gagtctgggt ataaagggag atgataagag gatttctcgc ctcatgaatc
3421 ctgcctttga tgagacagat gtagccgttc cagaaaagcc caaaactgat gccgaggttc
3481 gcatcgaggc agagaaagta tattacccac actatttcgt cgaaccccat aagaaggtgg
3541 aacgcgagga gaaaccctgt ggtcatcaaa agttccacga ggggcgactg acaggtaaaa
3601 ttcggtgtaa gctcattacc aagacacccc tcatcgtccc agatactagt aatgacgatt
3661 tcttcagacc tgcggataaa gaagctcgga aggaaaagga cgaatatcat aaatcatatg
3721 ctttcttcag acttcataaa caaatcatga ttcccgggag cgaattgaga ggaatggtga
3781 gtagtgtcta cgaaactgtg acaaattctt gcttcaggat atttgatgag actaaacggt
3841 tgtcatggcg gatggatgct gatcaccaga atgtgctgca agactttctc ccaggtcgag
3901 tgaccgccga tgggaaacat atacaaaagt tttccgaaac tgcaagggtg cctttctatg
3961 acaaaacgca gaaacatttt gacattcttg atgagcaaga aattgctggt gagaaacctg
4021 ttcggatgtg ggtcaagcgt tttattaaac gactgagcct cgttgatcct gctaaacacc
4081 cccagaagaa acaagacaat aaatggaaaa gacgcaaaga aggcatcgcc acatttatag
4141 agcaaaagaa tggctcttat tattttaacg tggtcaccaa taatggctgc acttctttcc
4201 acttgtggca taaacctgac aattttgacc aggagaaact cgaaggcata cagaatggtg
4261 agaaactgga ttgctgggta agagatagta gataccaaaa ggcctttcaa gagatacccg
4321 agaacgaccc agacggatgg gagtgtaaag agggctacct tcatgtcgtc ggccccagca
4381 aagtagagtt tagtgacaag aaaggggatg tgatcaataa ctttcaagga acactcccat
4441 cagttcccaa cgactggaaa acaattagga cgaacgattt taagaatagg aaaaggaaga
4501 atgaacctgt gttttgttgt gaggatgata agggcaatta ctataccatg gctaaatatt
4561 gcgaaacctt cttcttcgat ctgaaggaga acgaggaata cgagatcccc gagaaagcaa
4621 gaatcaaata caaagaactg ctcagggtct ataacaacaa tcctcaagca gtgccggaga
4681 gcgtatttca gtctagagtt gcccgggaaa acgtggaaaa gctgaagtcc ggagatcttg
4741 tgtatttcaa acataatgaa aagtacgtag aggacatcgt cccagtgcgg atttcccgaa
4801 ctgtagacga taggatgatc ggcaaacgta tgagcgccga tctgcggccg tgccatggag
4861 attgggtgga agatggtgat ctcagtgcct tgaatgcata tcccgagaaa agactcctct
4921 tgcgccaccc caaaggactc tgccctgctt gccggctctt tggaaccgga tcttacaagg
4981 gcagagtcag gtttggattc gcgtcactcg aaaacgatcc ggagtggctg atcccaggca
5041 agaatcccgg cgatccgttt cacggcgggc cggtgatgct ctcattgttg gaacggcctc
5101 gcccgacttg gagtataccg ggatccgaca ataagtttaa agtgcctggc agaaagtttt
5161 acgtccacca ccacgcctgg aaaaccatta aggacgggaa ccatcccaca acaggcaaag
5221 ctattgaaca aagccctaat aaccgcactg tagaagctct cgccggcggg aattccttta
5281 gcttcgaaat tgcctttgag aacctgaaag aatgggagct gggtttgctc atccacagcc
5341 tgcaactcga aaagggtctg gcgcataaac ttggaatggc aaagtctatg ggatttggtt
5401 cagttgaaat tgacgtcgaa tcagtgcgcc tgagaaaaga ttggaagcaa tggcggaatg
5461 gcaattccga aattcccaac tggttgggaa aaggatttgc taaactgaag gaatggttcc
5521 gggacgagct cgattttata gaaaatctta agaaacttct ttggtttcct gagggcgacc
5581 aagcaccccg ggtttgctac cccatgctgc gaaagaagga cgatcctaat gggaatagcg
5641 gttacgaaga actcaaagac ggggaattca agaaagaaga tcggcagaag aagctgacca
5701 cgccgtggac accgtgggca taaccggtca tcatcaccat caccattgag tttaaacccg
5761 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
5821 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgagaaaat
5881 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
5941 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
6001 ttctgaggcg gaaagaacca gctggggctc gataccgtcg acctctagct agagcttggc
6061 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
6121 catacgagcc ggaagcataa agtgtaaagc ctagggtgcc taatgagtga gctaactcac
6181 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
6241 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
6301 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
6361 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
6421 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
6481 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
6541 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
6601 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
6661 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
6721 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
6781 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
6841 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
6901 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
6961 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
7021 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
7081 tacggggtct gacactcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
7141 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
7201 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
7261 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
7321 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
7381 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
7441 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
7501 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
7561 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
7621 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
7681 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
7741 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
7801 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
7861 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
7921 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
7981 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
8041 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
8101 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
8161 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttcc
//
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