| 质粒名称: | pMA-SpCas9-g2 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 80785 |
| 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: | BsrGI (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-g2 was a gift from Yonglun Luo (Addgene plasmid # 80785 ; http://n2t.net/addgene:80785 ; RRID:Addgene_80785) |
| 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-g2 2789 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION BsaI-based cloning of SpCas9 gRNA guide sequence, position 2/12/22
in the array.
ACCESSION .
VERSION .
KEYWORDS pMA-SpCas9-g2
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 complement(215..232)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(325..913)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(405..424)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1084..1944)
/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 1707..1726
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1945..2049)
/gene="bla"
/label=AmpR promoter
primer_bind 2350..2372
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 2364..2381
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 2365..2381
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2392..2411
/label=T7
/note="T7 promoter, forward primer"
promoter 2392..2410
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 2475..2715
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 2475..2495
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 2646..2665
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 2742..28
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
ORIGIN
1 acttgaaaaa gtggcaccga gtcggtgctt ttttgtttta gagctagaaa tagcaagtta
61 ggaccgagac ctgtacaacg cgtggtacct ggagcacaag actggcctca cgggccttcc
121 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa catggtcata
181 gctgtttcct tgcgtattgg gcgctctccg cttcctcgct cactgactcg ctgcgctcgg
241 tcgttcgggt aaagcctggg gtgcctaatg agcaaaaggc cagcaaaagg ccaggaaccg
301 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
361 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
421 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
481 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
541 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
601 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
661 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
721 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
781 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
841 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
901 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
961 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
1021 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
1081 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
1141 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
1201 ccccagtgct gcaatgatac cgcgagaacc acgctcaccg gctccagatt tatcagcaat
1261 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
1321 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
1381 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
1441 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
1501 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
1561 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
1621 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
1681 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
1741 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
1801 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
1861 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
1921 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
1981 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
2041 ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta
2101 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
2161 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtggccg ctacagggcg
2221 ctcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgtttcggtg cgggcctctt
2281 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
2341 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagcgcgac gtaatacgac
2401 tcactatagg gcgaattggc ggaaggccgt caaggccacg tgtcttgtcc agagctcgga
2461 tccggtctcc catggagggc ctatttccca tgattccttc atatttgcat atacgataca
2521 aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata ttagtacaaa
2581 atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaag ttatgtttta
2641 aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata
2701 tcttgtggaa aggacgaaac accgggtctt cgagaagacc tgttttagag ctagaaatag
2761 caagttaaaa taaggctagt ccgttatca
//
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