| 质粒名称: | pT7-gRNA |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 46759 |
| 存储实验室: | Wenbiao Chen |
|
载体骨架基本信息 |
|
| 载体名称: | pUC19 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 2400 bp |
| 完整质粒大小: | 2540 bp |
| 载体修饰: | 删除253 bp AatII/NdeI片段 |
| 载体类型: | CRISPR,斑马鱼表达 |
| 筛选标记: | -- |
| 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: | gRNA core |
| 物种来源: | Synthetic |
| 插入大小: | 76 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | T7 |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | HindIII (在克隆过程中被破坏) |
| 3' 克隆位点: | BamHI(未破坏) |
| 5' 测序引物 | M13 R |
| 3' 测序引物 | M13 F |
|
文献引用方法 |
|
| 材料和方法部分: | pT7-gRNA was a gift from Wenbiao Chen (Addgene plasmid # 46759 ; http://n2t.net/addgene:46759 ; RRID:Addgene_46759) |
| 参考文献部分: | Efficient multiplex biallelic zebrafish genome editing using a CRISPR nuclease system. Jao LE, Wente SR, Chen W. Proc Natl Acad Sci U S A. 2013 Aug 5. 10.1073/pnas.1308335110 PubMed 23918387 |
买家导航
pT7gRNA载体基本信息
pT7gRNA载体图谱质粒图谱和多克隆位点信息
pT7gRNA载体简介
pT7gRNA载体序列
LOCUS pT7-gRNA 2541 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pT7-gRNA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2541)
AUTHORS Jao LE, Wente SR, Chen W
TITLE Efficient multiplex biallelic zebrafish genome editing using a
CRISPR nuclease system.
JOURNAL Proc Natl Acad Sci U S A. 2013 Aug 5.
PUBMED 23918387
REFERENCE 2 (bases 1 to 2541)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.biofeng.com/
FEATURES Location/Qualifiers
source 1..2541
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 180..202
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 194..211
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 195..211
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_RNA complement(239..314)
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
primer_bind complement(347..366)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(348..366)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(390..406)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(390..406)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(403..425)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 414..430
/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(438..468)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 483..504
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(621..638)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(792..1380)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(872..891)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1551..2411)
/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 2174..2193
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2412..2516)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
61 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
121 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
181 ccagtcacga cgttgtaaaa cgacggccag tgaattcgag ctcggtaccc ggggatccgc
241 accgactcgg tgccactttt tcaagttgat aacggactag ccttatttta acttgctatt
301 tctagctcta aaacagagac ggtcgactca gactagatct cgtctcccct atagtgagtc
361 gtattaattt caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
421 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
481 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
541 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
601 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
661 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
721 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
781 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
841 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
901 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
961 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
1021 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
1081 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
1141 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
1201 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
1261 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
1321 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
1381 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
1441 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
1501 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
1561 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
1621 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
1681 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
1741 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
1801 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
1861 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
1921 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
1981 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
2041 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
2101 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
2161 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
2221 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
2281 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
2341 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
2401 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
2461 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
2521 tttccccgaa aagtgccacc t
//
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