| 质粒名称: | pUC19-U6-AagRNA |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 121953 |
| 存储实验室: | Wei Li |
|
载体骨架基本信息 |
|
| 载体名称: | pUC19, BsaI site mutated |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 3135 bp |
| 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: |
AaCas12b tracrRNA/crRNA duplex、AaC2c1 tracrRNA/crRNA duplex
gRNA/shRNA
|
| 物种来源: | Synthetic |
| 克隆方法: | Ligation Independent Cloning |
|
文献引用方法 |
|
| 材料和方法部分: | pUC19-U6-AagRNA was a gift from Wei Li (Addgene plasmid # 121953 ; http://n2t.net/addgene:121953 ; RRID:Addgene_121953) |
| 参考文献部分: | Repurposing CRISPR-Cas12b for mammalian genome engineering. Teng F, Cui T, Feng G, Guo L, Xu K, Gao Q, Li T, Li J, Zhou Q, Li W. Cell Discov. 2018 Nov 27;4:63. doi: 10.1038/s41421-018-0069-3. eCollection 2018. 10.1038/s41421-018-0069-3 PubMed 30510770 |
买家导航
pUC19-U6-AagRNA载体质粒基本信息
pUC19-U6-AagRNA质粒图谱载体图谱和 pUC19-U6-AagRNA载体序列质粒序列多克隆位点信息
pUC19-U6-AagRNA质粒载体简介
pUC19-U6-AagRNA质粒序列载体序列
LOCUS pUC19-U6-AagRNA 3135 bp DNA circular SYN 15-APR-2025
DEFINITION Mammalian expression, Genome editing, gRNA scaffold.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3135)
AUTHORS Teng F, Cui T, Feng G, Guo L, Xu K, Gao Q, Li T, Li J, Zhou Q, Li W
TITLE Repurposing CRISPR-Cas12b for mammalian genome engineering.
JOURNAL Cell Discov. 2018 Nov 27;4:63. doi: 10.1038/s41421-018-0069-3.
eCollection 2018.
PUBMED 30510770
REFERENCE 2 (bases 1 to 3135)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 15, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell
Discov."; date: "2018-11-27"; volume: "4:63. doi"; pages: "
10.1038/s41421-018-0069-3. eCollection 2018"
COMMENT Sequence Label: pUC19-U6-AagRNA
FEATURES Location/Qualifiers
source 1..3135
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 62..84
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 76..93
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 77..93
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 100..348
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind 100..120
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 271..290
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
primer_bind complement(612..628)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(612..628)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(625..647)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 636..652
/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(660..666,667..684,685..690))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 705..726
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(843..860)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1014..1602)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1094..1113)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(1773..2564,2565..2633))
/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 2396..2415
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2634..2738)
/gene="bla"
/label=AmpR promoter
primer_bind 2806..2824
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(2862..2884)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 2984..3003
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
61 tcccagtcac gacgttgtaa aacgacggcc agtgaattcg agggcctatt tcccatgatt
121 ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat taatttgact
181 gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag
241 tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag
301 tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgg gtctaaagga
361 cagaattttt caacgggtgt gccaatggcc actttccagg tggcaaagcc cgttgaactt
421 ctcaaaaaga acgctcgctc agtgttctga caacaaagca ccagtggtct agtggtggaa
481 tagtaccctg ccacggtaca gacccgggtt cgattcccgg ctggtgcagt cggatcactg
541 agcgagcgat ctgagaagtg gcacagagac cgagagaggg tctcattttt tttaagcttg
601 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
661 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
721 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
781 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
841 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
901 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
961 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
1021 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
1081 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
1141 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
1201 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
1261 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
1321 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
1381 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
1441 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
1501 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
1561 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
1621 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
1681 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
1741 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
1801 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
1861 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagagcca
1921 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
1981 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
2041 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
2101 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
2161 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
2221 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
2281 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
2341 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
2401 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
2461 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
2521 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
2581 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
2641 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
2701 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
2761 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
2821 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
2881 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
2941 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
3001 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
3061 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
3121 gcctcttcgc tatta
//
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