| 质粒名称: | p3s-Sniper-Cas9 |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 113912 |
| 存储实验室: | Jungjoon Lee |
|
载体骨架基本信息 |
|
| 载体名称: | pCDNA3.1 |
| 载体出品公司: | invitrogen |
| 空载体大小: | 3200 bp |
| 完整质粒大小: | 7383 bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 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: | Sniper-Cas9 |
| 启动子: | CMV |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | CGCAAATGGGCGGTAGGCGTG |
| 3’测序引物 | TAGAAGGCACAGTCGAGG |
|
文献引用方法 |
|
| 材料和方法部分: | p3s-Sniper-Cas9 was a gift from Jungjoon Lee (Addgene plasmid # 113912 ; http://n2t.net/addgene:113912 ; RRID:Addgene_113912) |
| 参考文献部分: | Directed evolution of CRISPR-Cas9 to increase its specificity. Lee JK, Jeong E, Lee J, Jung M, Shin E, Kim YH, Lee K, Jung I, Kim D, Kim S, Kim JS. Nat Commun. 2018 Aug 6;9(1):3048. doi: 10.1038/s41467-018-05477-x. 10.1038/s41467-018-05477-x PubMed 30082838 |
买家导航
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 p3s-Sniper-Cas9 7383 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses Sniper-Cas9 with CMV promoter.
ACCESSION .
VERSION .
KEYWORDS p3s-Sniper-Cas9
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7383)
AUTHORS Lee JK, Jeong E, Lee J, Jung M, Shin E, Kim YH, Lee K, Jung I, Kim
D, Kim S, Kim JS
TITLE Directed evolution of CRISPR-Cas9 to increase its specificity.
JOURNAL Nat Commun. 2018 Aug 6;9(1):3048. doi: 10.1038/s41467-018-05477-x.
PUBMED 30082838
REFERENCE 2 (bases 1 to 7383)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7383
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 895..904
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 5020..5040
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5041..5067
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
promoter complement(5101..5119)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(5102..5119)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(5139..5156)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5145..5369
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(5457..5474)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5628..6216)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5708..5727)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6387..7247)
/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 7010..7029
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7248..7352)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgccacc
901 atggacaaga agtacagcat cggcctggac atcggtacca acagcgtggg ctgggccgtg
961 atcaccgacg agtacaaggt gcccagcaag aagttcaagg tgctgggcaa caccgaccgc
1021 cacagcatca agaagaacct gatcggcgcc ctgctgttcg acagcggcga gaccgccgag
1081 gccacccgcc tgaagcgcac cgcccgccgc cgctacaccc gccgcaagaa ccgcatctgc
1141 tacctgcagg agatcttcag caacgagatg gccaaggtgg acgacagctt cttccaccgc
1201 ctggaggaga gcttcctggt ggaggaggac aagaagcacg agcgccaccc catcttcggc
1261 aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgcgcaag
1321 aagctggtgg acagcaccga caaggccgac ctgcgcctga tctacctggc cctggcccac
1381 atgatcaagt tccgcggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac
1441 gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggagaacccc
1501 atcaacgcca gcggcgtgga cgccaaggcc atcctgagcg cccgcctgag caagagccgc
1561 cgcctggaga acctgatcgc ccagctgccc ggcgagaaga agaacggcct gttcggcaac
1621 ctgatcgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag
1681 gacgccaagc tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc
1741 cagatcggcg accagtacgc cgacctgttc ctggccgcca agaacctgag cgacgccatc
1801 ctgctgagcg acatcctgcg cgtgaacacc gagatcacca aggcccccct gagcgccagc
1861 atgatcaagc gctacgacga gcaccaccag gacctgaccc tgctgaaggc cctggtgcgc
1921 cagcagctgc ccgagaagta caaggagatc ttcttcgacc agagcaagaa cggctacgcc
1981 ggctacatcg acggcggcgc cagccaggag gagttctaca agttcatcaa gcccatcctg
2041 gagaagatgg acggcaccga ggagctgctg gtgaagctga accgcgagga cctgctgcgc
2101 aagcagcgca ccttcgacaa cggcagcatc ccccaccaga tccacctggg cgagctgcac
2161 gccatcctgc gccgccagga ggacttctac cccttcctga aggacaaccg cgagaagatc
2221 gagaagatcc tgaccttccg catcccctac tacgtgggcc ccctggcccg cggcaacagc
2281 cgcttcgcct ggatgacccg caagagcgag gagaccatca ccccctggaa cttcgaggag
2341 gtggtggaca agggcgccag cgcccagagc ttcatcgagc gcatgaccaa cttcgacaag
2401 aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg
2461 tacaacgagc tgaccaaggt gaagtacgtg accgagggca tgcgcaagcc cgcctccctg
2521 agcggcgagc agaagaaggc catcgtggac ctgctgttca agaccaaccg caaggtgacc
2581 gtgaagcagc tgaaggagga ctacttcaag aagatcgagt gcttcgacag cgtggagatc
2641 agcggcgtgg aggaccgctt caacgccagc ctgggcacct accacgacct gctgaagatc
2701 atcaaggaca aggacttcct ggacaacgag gagaacgagg acatcctgga ggacatcgtg
2761 ctgaccctga ccctgttcga ggaccgcgag atgatcgagg agcgcctgaa gacctacgcc
2821 cacctgttcg acgacaaggt gatgaagcag ctgaagcgcc gccgctacac cggctggggc
2881 cgcctgagcc gcaagcttat caacggcatc cgcgacaagc agagcggcaa gaccatcctg
2941 gacttcctga agagcgacgg cttcgccaac cgcaacttca tgcagctgat ccacgacgac
3001 agcctgacct tcaaggagga catccagaag gcccaggtga gcggccaggg cgacagcctg
3061 cacgagcaca tcgccaacct ggccggcagc cccgccatca agaagggcat cctgcagacc
3121 gtgaaggtgg tggacgagct ggtgaaggtg atgggccgcc acaagcccga gaacatcgtg
3181 atcgagatag cccgcgagaa ccagaccacc cagaagggcc agaagaacag ccgcgagcgc
3241 atgaagcgca tcgaggaggg catcaaggag ctgggcagcc agatcctgaa ggagcacccc
3301 gtggagaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaacggccgc
3361 gacatgtacg tggaccagga gctggacatc aaccgcctga gcgactacga cgtggaccac
3421 atcgtgcccc agagcttcct gaaggacgac agcatcgaca acaaggtgct gacccgcagc
3481 gacaagaacc gcggcaagag cgacaacgtg cccagcgagg aggtggtgaa gaagatgaag
3541 aactactggc gccagctgct gaacgccaat ctgatcaccc agcgcaagtt cgacaacctg
3601 accaaggccg agcgcggcgg cctgagcgag ctggacaagg ccggcttcat caagcgccag
3661 ctggtggaga cccgccagat caccaagcac gtggcccaga tcctggacag ccgcatgaac
3721 accaagtacg acgagaacga caagctgatc cgcgaggtga aggtgatcac cctgaagagc
3781 aagctggtga gcgacttccg caaggacttc cagttctaca aggtgcgcga gatcaacaac
3841 taccaccacg cccacgacgc ctacctgaac gccgtggtgg gcaccgccct gatcaagaag
3901 taccccaagc tggagagcga gttcgtgtac ggcgactaca aggtatacga cgtgcgcaag
3961 atgatcgcta agagcgagca ggagatcggc aaggccaccg ccaagtactt cttctacagc
4021 aacatcatga acttcttcaa gaccgagatc accctggcca acggcgagat ccgcaagcgc
4081 cccctgatcg agaccaacgg cgagaccggc gagatcgtgt gggacaaggg ccgcgacttc
4141 gccaccgtgc gcaaggtgct gagcatgccc caggtgaaca tcgtgaagaa gaccgaggtg
4201 cagaccggcg gcttcagcaa ggagagcatc ctgcccaagc gcaacagcga caagctgatc
4261 gcccgcaaga aggactggga ccccaagaag tacggcggct tcgacagccc caccgtggcc
4321 tacagcgtgc tggtggtggc caaggtggag aagggcaaga gcaagaagct gaagagcgtg
4381 aaggagctgc tgggcatcac catcatggag cgcagcagct tcgagaagaa ccccatcgac
4441 ttcctggagg ccaagggcta caaggaggtg aagaaggacc tgatcatcaa gctgcccaag
4501 tacagcctgt tcgagctgga gaacggccgc aagcgcatgc tggccagcgc cggcgagctg
4561 cagaagggca acgagctggc cctgcccagc aagtacgtga acttcctgta cctggccagc
4621 cactacgaga agctgaaggg cagccccgag gacaacgagc agaagcagct gttcgtggag
4681 cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttcag caagcgcgtg
4741 atcctggccg acgccaacct ggacaaggtg ctgagcgcct acaacaagca ccgcgacaag
4801 cccatccgcg agcaggccga gaacatcatc cacctgttca ccctgaccaa cctgggcgcc
4861 cccgccgcct tcaagtactt cgacaccacc atcgaccgca agcgctacac cagcaccaag
4921 gaggtgctgg acgccaccct gatccaccag agcatcaccg gtctgtacga gacccgcatc
4981 gacctgagcc agctgggcgg cgacggcggc tccggacctc caaagaaaaa gagaaaagta
5041 tacccctacg acgtgcccga ctacgcctaa taactcgagc atgcatctag agggccctat
5101 tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg
5161 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
5221 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
5281 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
5341 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctgcattaa
5401 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
5461 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
5521 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
5581 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
5641 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
5701 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
5761 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
5821 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
5881 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
5941 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
6001 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
6061 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
6121 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
6181 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
6241 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
6301 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
6361 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
6421 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
6481 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
6541 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
6601 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
6661 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
6721 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
6781 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
6841 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
6901 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
6961 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
7021 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
7081 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
7141 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
7201 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
7261 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
7321 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
7381 gtc
//
