| 质粒名称: | pMA-SpCas9-g7 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 80790 |
| 存储实验室: | Yonglun Luo |
|
载体骨架基本信息 |
|
| 载体名称: | pMA15 |
| 载体出品公司: | Lifetechnologies |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 2380 bp |
| 完整质粒大小: | 2789 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达.CRISPR |
| 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: | CRISPR gRNA expression cassette (for SpCas9) |
| 物种来源: | Synthetic |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 |
U6 |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BamHI(未破坏) |
| 3' 克隆位点: | BsmBI(未破坏) |
| 5' 测序引物 | TAATACGACTCACTATAGG |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pMA-SpCas9-g7 was a gift from Yonglun Luo (Addgene plasmid # 80790 ; http://n2t.net/addgene:80790 ; RRID:Addgene_80790) |
| 参考文献部分: | Golden Gate Assembly of CRISPR gRNA expression array for simultaneously targeting multiple genes. Vad-Nielsen J, Lin L, Bolund L, Nielsen AL, Luo Y. Cell Mol Life Sci. 2016 May 13. 10.1007/s00018-016-2271-5 [pii] PubMed 27178736 |
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载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pMA-SpCas9-g7 2789 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION BsaI-based cloning of SpCas9 gRNA guide sequence, position 7/17/27
in the array.
ACCESSION .
VERSION .
KEYWORDS pMA-SpCas9-g7
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2789)
AUTHORS Vad-Nielsen J, Lin L, Bolund L, Nielsen AL, Luo Y
TITLE Golden Gate Assembly of CRISPR gRNA expression array for
simultaneously targeting multiple genes.
JOURNAL Cell Mol Life Sci. 2016 May 13.
PUBMED 27178736
REFERENCE 2 (bases 1 to 2789)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..2789
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 438..457
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 630..647
/label=L4440
/note="L4440 vector, forward primer"
misc_RNA complement(834..909)
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter complement(936..1176)
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind complement(986..1005)
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
primer_bind complement(1156..1176)
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind complement(1240..1259)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(1241..1259)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1270..1287)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(1270..1286)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1279..1301)
/label=M13/pUC Forward
/note="In lacZ gene"
promoter 1602..1706
/gene="bla"
/label=AmpR promoter
CDS 1707..2567
/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(1925..1944)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2738..537
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
61 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
121 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
181 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
241 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
301 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
361 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
421 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
481 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
541 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcattaggca
601 ccccaggctt tacccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggagagcg
661 cccaatacgc aaggaaacag ctatgaccat gttaatgcag ctggcacgac aggtttcccg
721 actggaaagc gggcagtgag cggaaggccc gtgaggccag tcttgtgctc caggtaccac
781 gcgttgtaca ggtctcgttt ctaacttgct atttctagct ctaaaacaaa aaagcaccga
841 ctcggtgcca ctttttcaag ttgataacgg actagcctta ttttaacttg ctatttctag
901 ctctaaaaca ggtcttctcg aagacccggt gtttcgtcct ttccacaaga tatataaagc
961 caagaaatcg aaatactttc aagttacggt aagcatatga tagtccattt taaaacataa
1021 ttttaaaact gcaaactacc caagaaatta ttactttcta cgtcacgtat tttgtactaa
1081 tatctttgtg tttacagtca aattaattcc aattatctct ctaacagcct tgtatcgtat
1141 atgcaaatat gaaggaatca tgggaaatag gccctcaccg tgagaccgga tccgagctct
1201 ggacaagaca cgtggccttg acggccttcc gccaattcgc cctatagtga gtcgtattac
1261 gtcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
1321 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
1381 caccgaaacg cccttcccaa cagttgcgca gcctgaatgg cgaatgggag cgccctgtag
1441 cggccactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
1501 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
1561 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
1621 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
1681 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
1741 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
1801 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
1861 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
1921 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
1981 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
2041 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
2101 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
2161 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
2221 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
2281 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
2341 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
2401 aatctggagc cggtgagcgt ggttctcgcg gtatcattgc agcactgggg ccagatggta
2461 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
2521 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
2581 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
2641 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
2701 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
2761 taatctgctg cttgcaaaca aaaaaacca
//
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