| 出品公司: | -- |
|---|---|
| 载体名称: | pCasSA |
| 质粒类型: | 金黄色葡萄球菌基因敲除载体；金葡菌基因敲除载体；金葡菌/大肠杆菌穿梭质粒；金葡菌Crispr/cas9基因敲除载体；金黄色葡萄球菌Crispr/cas9基因敲除载体; 金黄色葡萄球菌Crispr载体 |
| 高拷贝/低拷贝: | 高拷贝 |
| 克隆方法: | 限制性内切酶，多克隆位点 |
| 启动子: | -- |
| 载体大小: | 10.2 kb |
| 5' 测序引物及序列: | M13-F: TGTAAAACGACGGCCAGT |
| 3' 测序引物及序列: | M13-R: CAGGAAACAGCTATGAC |
| 载体抗性: | 卡那青霉素 |
| 筛选标记: | 氯霉素 |
| 克隆菌株: | DH5α 等 |
| 宿主细胞（系）: | 金黄色葡萄球菌 |
| 备注: | 本载体是使用Crispr-Cas9技术进行金黄色葡萄球菌基因敲除的载体质粒，本质粒是大肠杆菌-金葡菌穿梭质粒，用于金葡菌基因敲除。详细实验方案见下面的参考文献！ |
| 产品目录号: | -- |
| 稳定性: | 稳表达 |
| 组成型/诱导型: | 组成型 |
| 病毒/非病毒: | 非病毒 |
| 参考文献： | Rapid and Efficient Genome Editing in Staphylococcus aureus by Using an Engineered CRISPR/Cas9 System. Chen W, Zhang Y, Yeo WS, Bae T, Ji Q. J Am Chem Soc. 2017 Mar 15;139(10):3790-3795. doi: 10.1021/jacs.6b13317. Epub 2017 Mar 2. 10.1021/jacs.6b13317 PubMedID: 28218837 |
买家导航
pCasSA载体载体Crispr-Cas9金黄色葡萄球菌金葡菌基因敲除质粒基本信息
pCasSA质粒图谱载体Crispr-Cas9金黄色葡萄球菌金葡菌基因敲除载体图谱和pCasSA载体序列载体Crispr-Cas9金黄色葡萄球菌金葡菌基因敲除质粒序列多克隆位点信息
pCasSA质粒载体Crispr-Cas9金黄色葡萄球菌金葡菌基因敲除载体简介
pCasSA质粒序列载体Crispr-Cas9金黄色葡萄球菌金葡菌基因敲除载体序列
LOCUS pCasSA 10240 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION CRISPR-based E. coli/Staphylococcus aureus temperature-sensitive
plasmid for genome editing in Staphylococcus aureus.
ACCESSION .
VERSION .
KEYWORDS pCasSA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10240)
AUTHORS Chen W, Zhang Y, Yeo WS, Bae T, Ji Q
TITLE Rapid and Efficient Genome Editing in Staphylococcus aureus by Using
an Engineered CRISPR/Cas9 System.
JOURNAL J Am Chem Soc. 2017 Mar 15;139(10):3790-3795. doi:
10.1021/jacs.6b13317. Epub 2017 Mar 2.
PUBMED 28218837
REFERENCE 2 (bases 1 to 10240)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..10240
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 79..667
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 568..587
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
misc_RNA 805..880
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
CDS 1121..5227
/codon_start=1
/product="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
/label=Cas9
/note="generates RNA-guided double strand breaks in DNA"
primer_bind 5324..5342
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(5380..5402)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 5502..5521
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5715..5737
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 5729..5746
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 5730..5746
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 9125..9141
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 9125..9141
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
CDS 9312..10106
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
primer_bind complement(9366..9385)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 9976..9995
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
terminator 10137..10231
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
ORIGIN
1 agtcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
61 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
121 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
181 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
241 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
301 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
361 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
421 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
481 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
541 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
601 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
661 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
721 tcacatgttc tttcctgcgt tgtcgacaga gtttgcaaaa tatacagggg attatatata
781 atggaaacga gaccattggt ctcagtttta gagctagaaa tagcaagtta aaataaggct
841 agtccgttat caacttgaaa aagtggcacc gagtcggtgc tttttttgag atctgtccat
901 acccatggtc tagaatgctc gagtcagaaa aatatacctg tatctttttt caaaagcaaa
961 catgctttgg gtaaacatgt aggtattaac gtcaatgcga caatagtagc attgattaaa
1021 tgagaattag taagtgtttt acttactaaa ttttatttaa cctaaaaatg aaccacctgg
1081 atgtgtggga ttaaaaagtg aagagaggag gacatatcac atggataaga aatactcaat
1141 aggcttagat atcggcacaa atagcgtcgg atgggcggtg atcactgatg aatataaggt
1201 tccgtctaaa aagttcaagg ttctgggaaa tacagaccgc cacagtatca aaaaaaatct
1261 tataggggct cttttatttg acagtggaga gacagcggaa gcgactcgtc tcaaacggac
1321 agctcgtaga aggtatacac gtcggaagaa tcgtatttgt tatctacagg agattttttc
1381 aaatgagatg gcgaaagtag atgatagttt ctttcatcga cttgaagagt cttttttggt
1441 ggaagaagac aagaagcatg aacgtcatcc tatttttgga aatatagtag atgaagttgc
1501 ttatcatgag aaatatccaa ctatctatca tctgcgaaaa aaattggtag attctactga
1561 taaagcggat ttgcgcttaa tctatttggc cttagcgcat atgattaagt ttcgtggtca
1621 ttttttgatt gagggagatt taaatcctga taatagtgat gtggacaaac tatttatcca
1681 gttggtacaa acctacaatc aattatttga agaaaaccct attaacgcaa gtggagtaga
1741 tgctaaagcg attctttctg cacgattgag taaatcaaga cgattagaaa atctcattgc
1801 tcagctcccc ggtgagaaga aaaatggctt atttgggaat ctcattgctt tgtcattggg
1861 tttgacccct aattttaaat caaattttga tttggcagaa gatgctaaat tacagctttc
1921 aaaagatact tacgatgatg atttagataa tttattggcg caaattggag atcaatatgc
1981 tgatttgttt ttggcagcta agaatttatc agatgctatt ttactttcag atatcctaag
2041 agtaaatact gaaataacta aggctcccct atcagcttca atgattaaac gctacgatga
2101 acatcatcaa gacttgactc ttttaaaagc tttagttcga caacaacttc cagaaaagta
2161 taaagaaatc ttttttgatc aatcaaaaaa cggatatgca ggttatattg atgggggagc
2221 tagccaagaa gaattttata aatttatcaa accaatttta gaaaaaatgg atggtactga
2281 ggaattattg gtgaaactaa atcgtgaaga tttgctgcgc aagcaacgga cctttgacaa
2341 cggctctatt ccccatcaaa ttcacttggg tgagctgcat gctattttga gaagacaaga
2401 agacttttat ccatttttaa aagacaatcg tgagaagatt gaaaaaatct tgacttttcg
2461 aattccttat tatgttggtc cattggcgcg tggcaatagt cgttttgcat ggatgactcg
2521 gaagtctgaa gaaacaatta ccccatggaa ttttgaagaa gttgtcgata aaggtgcttc
2581 agctcaatca tttattgaac gcatgacaaa ctttgataaa aatcttccaa atgaaaaagt
2641 actaccaaaa catagtttgc tttatgagta ttttacggtt tataacgaat tgacaaaggt
2701 caaatatgtt actgaaggaa tgcgaaaacc agcatttctt tcaggtgaac agaagaaagc
2761 cattgttgat ttactcttca aaacaaatcg aaaagtaacc gttaagcaat taaaagaaga
2821 ttatttcaaa aaaatagaat gttttgatag tgttgaaatt tcaggagttg aagatagatt
2881 taatgcttca ttaggtacct accatgattt gctaaaaatt attaaagata aagatttttt
2941 ggataatgaa gaaaatgaag atatcttaga ggatattgtt ttaacattga ccttatttga
3001 agatagggag atgattgagg aaagacttaa aacatatgct cacctctttg atgataaggt
3061 gatgaaacag cttaaacgtc gccgttatac tggttgggga cgtttgtctc gaaaattgat
3121 taatggtatt agggataagc aatctggcaa aacaatatta gattttttga aatcagatgg
3181 ttttgccaat cgcaatttta tgcagctgat ccatgatgat agtttgacat ttaaagaaga
3241 cattcaaaaa gcacaagtgt ctggacaagg cgatagttta catgaacata ttgcaaattt
3301 agctggtagc cctgctatta aaaaaggtat tttacagact gtaaaagttg ttgatgaatt
3361 ggtcaaagta atggggcggc ataagccaga aaatatcgtt attgaaatgg cacgtgaaaa
3421 tcagacaact caaaagggcc agaaaaattc gcgagagcgt atgaaacgaa tcgaagaagg
3481 tatcaaagaa ttaggaagtc agattcttaa agagcatcct gttgaaaata ctcaattgca
3541 aaatgaaaag ctctatctct attatctcca aaatggaaga gacatgtatg tggaccaaga
3601 attagatatt aatcgtttaa gtgattatga tgtcgatcac attgttccac aaagtttcct
3661 taaagacgat tcaatagaca ataaggtctt aacgcgttct gataaaaatc gtggtaaatc
3721 ggataacgtt ccaagtgaag aagtagtcaa aaagatgaaa aactattgga gacaacttct
3781 aaacgccaag ttaatcactc aacgtaagtt tgataattta acgaaagctg aacgtggagg
3841 tttgagtgaa cttgataaag ctggttttat caaacgccaa ttggttgaaa ctcgccaaat
3901 cactaagcat gtggcacaaa ttttggatag tcgcatgaat actaaatacg atgaaaatga
3961 taaacttatt cgagaggtta aagtgattac cttaaaatct aaattagttt ctgacttccg
4021 aaaagatttc caattctata aagtacgtga gattaacaat taccatcatg cccatgatgc
4081 gtatctaaat gccgtcgttg gaactgcttt gattaagaaa tatccaaaac ttgaatcgga
4141 gtttgtctat ggtgattata aagtttatga tgttcgtaaa atgattgcta agtctgagca
4201 agaaataggc aaagcaaccg caaaatattt cttttactct aatatcatga acttcttcaa
4261 aacagaaatt acacttgcaa atggagagat tcgcaaacgc cctctaatcg aaactaatgg
4321 ggaaactgga gaaattgtct gggataaagg gcgagatttt gccacagtgc gcaaagtatt
4381 gtccatgccc caagtcaata ttgtcaagaa aacagaagta cagacaggcg gattctccaa
4441 ggagtcaatt ttaccaaaaa gaaattcgga caagcttatt gctcgtaaaa aagactggga
4501 tccaaaaaaa tatggtggtt ttgatagtcc aacggtagct tattcagtcc tagtggttgc
4561 taaggtggaa aaagggaaat cgaagaagtt aaaatccgtt aaagagttac tagggatcac
4621 aattatggaa agaagttcct ttgaaaaaaa tccgattgac tttttagaag ctaaaggata
4681 taaggaagtt aaaaaagact taatcattaa actacctaaa tatagtcttt ttgagttaga
4741 aaacggtcgt aaacggatgc tggctagtgc cggagaatta caaaaaggaa atgagctggc
4801 tctgccaagc aaatatgtga attttttata tttagctagt cattatgaaa agttgaaggg
4861 tagtccagaa gataacgaac aaaaacaatt gtttgtggag cagcataagc attatttaga
4921 tgagattatt gagcaaatca gtgaattttc taagcgtgtt attttagcag atgccaattt
4981 agataaagtt cttagtgcat ataacaaaca tagagacaaa ccaatacgtg aacaagcaga
5041 aaatattatt catttattta cgttgacgaa tcttggagct cccgctgctt ttaaatattt
5101 tgatacaaca attgatcgta aacgatatac gtctacaaaa gaagttttag atgccactct
5161 tatccatcaa tccatcactg gtctttatga aacacgcatt gatttgagtc agctaggagg
5221 tgactgatgg ctggttggcg tactgttgtg gtaaataccc atgggtatgg acagttttcc
5281 cgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
5341 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
5401 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
5461 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
5521 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
5581 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
5641 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
5701 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca agcttgcatg
5761 cctgcagaac ggattgttga tgattacgaa aatattaaga gcacagacta ttacacagaa
5821 aatcaagaat taaaaaaacg tagagagagt ttgaaagaag tagtgaatac atggaaagag
5881 gggtatcacg aaaaaagtaa agaggttaat aaattaaagc gagagaatga tagtttgaat
5941 gagcagttga atgtatcaga gaaatttcaa gatagtacag tgactttata tcgtgctgcg
6001 agggcgaatt tccctgggtt tgagaaaggg tttaataggc ttaaagagaa attctttaat
6061 gattccaaat tcgagcgtgt gggacagttt atggatgttg tacaggataa tgtccagaag
6121 gtcgatagaa agcgtgagaa acagcgtaca gacgatttag agatgtagag gtacttttat
6181 gccgagaaaa ctttttgcgt gtgacagtcc ttaaaatata cttagagcgt aagcgaaagt
6241 agtagcgaca gctattaact ttcggttgca aagctctagg atttttaatg gacgcagcgc
6301 atcacacgca aaaaggaaat tggaataaat gcgaaatttg agatgttaat taaagacctt
6361 tttgaggtct ttttttctta gatttttggg gttatttagg ggagaaaaca taggggggta
6421 ctacgacctc ccccctaggt gtccattgtc cattgtccaa acaaataaat aaatattggg
6481 tttttaatgt taaaaggttg ttttttatgt taaagtgaaa aaaacagatg ttgggaggta
6541 cagtgatggt tgtagataga aaagaagaga aaaaagttgc tgttacttta agacttacaa
6601 cagaagaaaa tgagatatta aatagaatca aagaaaaata taatattagc aaatcagatg
6661 caaccggtat tctaataaaa aaatatgcaa aggaggaata cggtgcattt taaacaaaaa
6721 aagatagaca gcactggcat gctgcctatc tatgactaaa ttttgttaaa tgtattagca
6781 ccgttattat atcatgagcg aaaatgtaat aaaagaaact gaaaacaaga aaaattcaag
6841 aggacgtaat tggacatttg ttttatatcc agaatcagca aaagccgagt ggttagagta
6901 tttaaaagag ttacacattc aatttgtagt gtctccatta catgataggg atactgatac
6961 agaagatagg atgaaaaaag agcattatca tattctagtg atgtatgagg gtaataaatc
7021 ttatgaacag ataaaaataa ttacagaaga attgaatgcg actattccgc agattgcagg
7081 aagtgtgaaa ggtcttgtga gatatatgct tcacatggac gatcctaata aatttaaata
7141 tcaaaaagaa gatatgatag tttatggcgg tgtagatgtt gatgaattat taaagaaaac
7201 aacaacagat agatataaat taattaaaga aatgattgag tttattgatg aacaaggaat
7261 cgtagaattt aagagtttaa tggattatgc aatgaagttt aaatttgatg attggttccc
7321 gcttttatgt gataactcgg cgtatgttat tcaagaatat ataaaatcaa atcggtataa
7381 atctgaccga tagattttga atttaagagt gtcacaagac actctttttt cgcaccagcg
7441 aaaactggtt taagccgact gcgcaaaaga cataatcgat tcacaaaaaa taggcacacg
7501 aaaaacaagt taagggatgc agtttatgca tcccttaact tacttattaa ataatttata
7561 gctattgaaa agagataaga attgttcaaa gctaatattg tttaaatcgt caattcctgc
7621 atgttttaag gaattgttaa attgattttt tgtaaatatt ttcttgtatt ctttgttatc
7681 ttggttaccc gtcttcttaa tatgcgtaat tgcatctact ctaaatccat caatgccttt
7741 atcaaaccac cagttcatca tttcaaatac agcatctcta acttccggat taccccaatt
7801 caaatcaggt tgttttttac tgaataaatg gaaataatat tgctcagtat tagcatcata
7861 ttcccgcgcg ccaattgagc tcccctttct atgtatgttt tttactagtc atttaaaacg
7921 atacattaat aggtacgaaa aagcaacttt ttttgcgctt aaaaccagtc ataccaataa
7981 cttaagggta actagcctcg ccggcaatag ttacccttat tatcaagata agaaagaaaa
8041 ggatttttcg ctacgctcaa atcctttaaa aaaacacaaa agaccacatt ttttaatgtg
8101 gtcttttatt cttcaactaa agcacccatt agttcaacaa acgaaaattg gataaagtgg
8161 gatattttta aaatatatat ttatgttaca gtaatattga cttttaaaaa aggattgatt
8221 ctaatgaaga aagcagacaa gtaagcctcc taaattcact ttagataaaa atttaggagg
8281 catatcaaat gaaatttaat aaaattgatt tagacaattg gaagagaaaa gagatattta
8341 atcattattt gaaccaacaa acgactttta gtataaccac agaaattgat attagtgttt
8401 tataccgaaa cataaaacaa gaaggatata aattttaccc tgcatttatt ttcttagtga
8461 caagggtgat aaactcaaat acagctttta gaactggtta caatagcgac ggagagttag
8521 gttattggga taagttagag ccactttata caatttttga tggtgtatct aaaacattct
8581 ctggtatttg gactcctgta aagaatgact tcaaagagtt ttatgattta tacctttctg
8641 atgtagagaa atataatggt tcggggaaat tgtttcccaa aacacctata cctgaaaatg
8701 ctttttctct ttctattatt ccatggactt catttactgg gtttaactta aatatcaata
8761 ataatagtaa ttaccttcta cccattatta cagcaggaaa attcattaat aaaggtaatt
8821 caatatattt accgctatct ttacaggtac atcattctgt ttgtgatggt tatcatgcag
8881 gattgtttat gaactctatt caggaattgt cagataggcc taatgactgg cttttataat
8941 atgagataat gccgactgta ctttttacag tcggttttct aatgtcacta acctgccccg
9001 ttagttgaag aaggttttta tattacagct ccagatccat atccttcttt ttctgaaccg
9061 acttctcctt tttcgcttct ttattccaat tgctttattg acgttgagcc tcggaaccgg
9121 tacccaggaa acagctatga ccatgtaata cgactcacta tacggggata tcgtcggaat
9181 tgccagctgg ggcgccctct ggtaaggttg ggaagccctg caaagtaaac tggatggctt
9241 tcttgccgcc aaggatctga tggcgcaggg gatcaagatc tgatcaagag acaggatgag
9301 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
9361 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
9421 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
9481 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
9541 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
9601 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
9661 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
9721 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
9781 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
9841 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
9901 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
9961 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
10021 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
10081 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgagagct cgcttggact
10141 cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca gaacgctcgg
10201 ttgccgccgg gcgtttttta ttggtgagaa tccaagcact
//
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