| 质粒名称: | PX853 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 62885 |
| 存储实验室: | Feng Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | PX330 |
| 载体出品公司: | Zhang lab |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4222bp |
| 完整质粒大小: | 6292 bp |
| 载体修饰: | 人细胞色素c氧化酶亚基VIII（human cytochrome C oxidase subunit VIII） |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 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: | SpCas9 (aa 2-573) |
| 物种来源: | -- |
| 插入大小: | 2073bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | PX853 was a gift from Feng Zhang (Addgene plasmid # 62885 ; http://n2t.net/addgene:62885 ; RRID:Addgene_62885) |
| 参考文献部分: | A split-Cas9 architecture for inducible genome editing and transcription modulation. Zetsche B, Volz SE, Zhang F. Nat Biotechnol. 2015 Feb 2. doi: 10.1038/nbt.3149. 10.1038/nbt.3149 PubMed 25643054 |
买家导航
PX853载体质粒基本信息
PX853质粒图谱载体图谱和PX853载体序列质粒序列多克隆位点信息
PX853质粒载体简介
PX853质粒序列载体序列
LOCUS PX853 6295 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION N-term SpCas9 piece of inducible split-5 (Cas9(N)-FRB-NES split-5)..
ACCESSION .
VERSION .
KEYWORDS PX853
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6295)
AUTHORS Zetsche B, Volz SE, Zhang F
TITLE A split-Cas9 architecture for inducible genome editing and
transcription modulation.
JOURNAL Nat Biotechnol. 2015 Feb 2. doi: 10.1038/nbt.3149.
PUBMED 25643054
REFERENCE 2 (bases 1 to 6295)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6295
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..241
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 1..21
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 172..191
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 268..343
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
enhancer 440..725
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer;
contains an 18-bp deletion relative to the standard CMV
enhancer"
promoter 727..1004
/label=chicken beta-actin promoter
intron 1005..1233
/label=hybrid intron
/note="hybrid between chicken beta-actin (CBA) and minute
virus of mice (MMV) introns (Gray et al., 2011)"
CDS 1278..2993
/codon_start=1
/product="N-terminal portion of Streptococcus pyogenes Cas9
(Zetsche et al., 2015)"
/label=Cas9(N)
/translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
CDS 3039..3308
/codon_start=1
/product="FKBP-rapamycin binding domain of human FRAP"
/label=FRB
/translation="EMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETS
FNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISKQ"
primer_bind complement(3336..3353)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3342..3549
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 3558..3698
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 3558..3687
/label=AAV2 ITR
rep_origin 3773..4228
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3860..3879)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4070..4091
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4245..4264)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4364..4386
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4424..4442)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4510..4614
/gene="bla"
/label=AmpR promoter
CDS 4615..5475
/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(4833..4852)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5646..6234
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6135..6154
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
241 cgaaacaccg ggtcttcgag aagacctgtt ttagagctag aaatagcaag ttaaaataag
301 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg ttttagagct
361 agaaatagca agttaaaata aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac
421 aaatggctct agaggtaccc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
481 ccaacgaccc ccgcccattg acgtcaatag taacgccaat agggactttc cattgacgtc
541 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
601 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tgtgcccagt
661 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
721 ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac
781 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg
841 ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga
901 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
961 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgacgc
1021 tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg
1081 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag
1141 ctgagcaaga ggtaagggtt taagggatgg ttggttggtg gggtattaat gtttaattac
1201 ctggagcacc tgcctgaaat cacttttttt caggttggac cggtgccacc atgctagatt
1261 tagctagctt aatcctcgac aagaagtaca gcatcggcct ggacatcggc accaactctg
1321 tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg
1381 gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg
1441 gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga
1501 agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca
1561 gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc
1621 accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct
1681 accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc
1741 tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc
1801 ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt
1861 tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac
1921 tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg
1981 gcctgttcgg aaacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact
2041 tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg
2101 acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc
2161 tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc
2221 ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga
2281 aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca
2341 agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca
2401 tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag
2461 aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc
2521 tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca
2581 accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg
2641 ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct
2701 ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga
2761 ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg
2821 agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa
2881 agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca
2941 accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc gagggtggcg
3001 gtggctcggg tggcggtggc tcgggtggcg gtggctcgga gatgtggcac gagggcctgg
3061 aggaggccag cagactgtac ttcggcgaga gaaacgtgaa gggcatgttc gaggtgctgg
3121 agcccctgca cgccatgatg gagagaggcc cccagaccct gaaggagacc agcttcaacc
3181 aggcctacgg cagagacctg atggaggccc aggagtggtg cagaaagtac atgaagagcg
3241 gcaacgtgaa ggacctgacc caggcctggg acctgtacta ccacgtgttc agaagaatca
3301 gcaagcagta agaattccta gagctcgctg atcagcctcg actgtgcctt ctagttgcca
3361 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
3421 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
3481 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaga atagcaggca
3541 tgctggggag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
3601 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
3661 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
3721 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
3781 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
3841 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
3901 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
3961 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
4021 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
4081 ttccaaactg gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt
4141 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
4201 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
4261 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
4321 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
4381 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
4441 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
4501 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
4561 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
4621 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
4681 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
4741 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
4801 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
4861 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
4921 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
4981 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
5041 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
5101 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
5161 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
5221 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
5281 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg aagccgcggt
5341 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
5401 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
5461 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
5521 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
5581 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
5641 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
5701 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
5761 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
5821 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
5881 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
5941 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
6001 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
6061 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
6121 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
6181 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
6241 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
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