| 質粒名稱: | pMA-SpCas9-g9 |
|---|---|
| 出品公司: | Addgene |
| 目錄編號: | 80792 |
| 存儲實驗室: | Yonglun Luo |
|
載體骨架基本信息 |
|
| 載體名稱: | pMA15 |
| 載體出品公司: | Lifetechnologies |
| 載體抗性: | 氨芐青霉素 |
| 空載體大小: | 2380 bp |
| 完整質粒大小: | 2789 bp |
| 載體修飾: | -- |
| 載體類型: | 哺乳細胞表達,CRISPR |
| 篩選標記: | -- |
| 高拷貝/低拷貝: | 高拷貝 |
| 生長條件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名稱: | CRISPR gRNA expression cassette (for SpCas9) |
| 物種來源: | Synthetic |
| 插入大小: | -- |
| 基因突變: | -- |
| GenBank編號 | -- |
| 啟動子 | U6 |
| 標簽/融合蛋白: | -- |
| 克隆方法: | 限制性內切酶 |
| 5' 克隆位點: | BamHI(未破壞) |
| 3' 克隆位點: | BsrGI (未破壞) |
| 5' 測序引物 | TAATACGACTCACTATAGG |
| 3' 測序引物 | -- |
|
文獻引用方法 |
|
| 材料和方法部分: | pMA-SpCas9-g9 was a gift from Yonglun Luo (Addgene plasmid # 80792 ; //n2t.net/addgene:80792 ; RRID:Addgene_80792) |
| 參考文獻部分: | 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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pMA-SpCas9-g9載體質粒基本信息
pMA-SpCas9-g9質粒圖譜載體圖譜和pMA-SpCas9-g9載體序列質粒序列多克隆位點信息
pMA-SpCas9-g9質粒載體簡介
pMA-SpCas9-g9質粒序列載體序列
LOCUS pMA-SpCas9-g9 2789 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION BsaI-based cloning of SpCas9 gRNA guide sequence, position 9/19/29
in the array.
ACCESSION .
VERSION .
KEYWORDS pMA-SpCas9-g9
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
//www.snapgene.com
FEATURES Location/Qualifiers
source 1..2789
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 278..300
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 292..309
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 293..309
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 320..339
/label=T7
/note="T7 promoter, forward primer"
promoter 320..338
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 403..643
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 403..423
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 574..593
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 670..745
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
primer_bind complement(932..949)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1042..1630)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1122..1141)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1801..2661)
/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 2424..2443
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2662..2766)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
121 gatagggttg agtggccgct acagggcgct cccattcgcc attcaggctg cgcaactgtt
181 gggaagggcg tttcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
241 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
301 acggccagtg agcgcgacgt aatacgactc actatagggc gaattggcgg aaggccgtca
361 aggccacgtg tcttgtccag agctcggatc cggtctcctc gagagggcct atttcccatg
421 attccttcat atttgcatat acgatacaag gctgttagag agataattgg aattaatttg
481 actgtaaaca caaagatatt agtacaaaat acgtgacgta gaaagtaata atttcttggg
541 tagtttgcag ttttaaaatt atgttttaaa atggactatc atatgcttac cgtaacttga
601 aagtatttcg atttcttggc tttatatatc ttgtggaaag gacgaaacac cgggtcttcg
661 agaagacctg ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact
721 tgaaaaagtg gcaccgagtc ggtgcttttt tgttttagag ctagaaatag caagttagct
781 cggagacctg tacaacgcgt ggtacctgga gcacaagact ggcctcacgg gccttccgct
841 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaacat ggtcatagct
901 gtttccttgc gtattgggcg ctctccgctt cctcgctcac tgactcgctg cgctcggtcg
961 ttcgggtaaa gcctggggtg cctaatgagc aaaaggccag caaaaggcca ggaaccgtaa
1021 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
1081 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
1141 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
1201 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
1261 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
1321 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
1381 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
1441 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
1501 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
1561 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
1621 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
1681 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
1741 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
1801 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
1861 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
1921 cagtgctgca atgataccgc gagaaccacg ctcaccggct ccagatttat cagcaataaa
1981 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
2041 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
2101 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
2161 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
2221 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
2281 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
2341 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
2401 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
2461 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
2521 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
2581 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
2641 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
2701 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
2761 tccgcgcaca tttccccgaa aagtgccac
//
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