质粒名称: | pSGKP-km |
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出品公司: | 爱迪基因 |
目录编号: | 117233 |
存储实验室: | Quanjiang Ji |
载体骨架基本信息 |
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载体名称: | -- |
载体出品公司: | -- |
载体抗性: | 卡那霉素 |
空载体大小: | 4729 bp |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | 细菌表达,CRISPR,合成生物学 |
筛选标记: | -- |
高拷贝/低拷贝: | 高拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | -- |
物种来源: | -- |
插入大小: | -- |
基因突变: | -- |
GenBank编号 | -- |
启动子 | -- |
标签/融合蛋白: | -- |
克隆方法: | Gibson克隆 |
5' 克隆位点: | -- |
3' 克隆位点: | -- |
5' 测序引物 | -- |
3' 测序引物 | -- |
文献引用方法 |
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材料和方法部分: | pSGKP-km was a gift from Quanjiang Ji (Addgene plasmid # 117233 ; http://n2t.net/addgene:117233 ; RRID:Addgene_117233) |
参考文献部分: | Precise and efficient genome editing in Klebsiella pneumoniae using CRISPR-Cas9 and CRISPR-assisted cytidine deaminase. Wang Y, Wang S, Chen W, Song L, Zhang Y, Shen Z, Yu F, Li M, Ji Q. Appl Environ Microbiol. 2018 Sep 14. pii: AEM.01834-18. doi: 10.1128/AEM.01834-18. 10.1128/AEM.01834-18 PubMed 30217854 |
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pSGKPkm载体基本信息
pSGKPkm载体图谱质粒图谱和多克隆位点信息
pSGKPkm载体简介
pSGKPkm载体序列
LOCUS pSGKP-km 4729 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION A sgRNA expression plasmid for genome editing in Klebsiella Pneumoniae. ACCESSION . VERSION . KEYWORDS pSGKP-km SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4729) AUTHORS Wang Y, Wang S, Chen W, Song L, Zhang Y, Shen Z, Yu F, Li M, Ji Q TITLE Precise and efficient genome editing in Klebsiella pneumoniae using CRISPR-Cas9 and CRISPR-assisted cytidine deaminase. JOURNAL Appl Environ Microbiol. 2018 Sep 14. pii: AEM.01834-18. doi: 10.1128/AEM.01834-18. PUBMED 30217854 REFERENCE 2 (bases 1 to 4729) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4729 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 97..131 /label=J23119(SpeI) promoter /note="bacterial promoter (Registry of Standard Biological Parts BBa_J23119) modified to end with an SpeI site" misc_RNA 149..224 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" primer_bind complement(257..273) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(257..273) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(332..348) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(332..348) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(345..367) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 356..372 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(380..410) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 425..446 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(563..580) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(734..1322) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(814..833) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1493..2308) /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind 2216..2235 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" promoter complement(2309..2413) /gene="bla" /label=AmpR promoter primer_bind 2481..2499 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(2537..2559) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 2872..2894 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2886..2903 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 2887..2903 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 2940..2956 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 2941..2957 /label=KS primer /note="common sequencing primer, one of multiple similar variants" CDS complement(3019..4440) /codon_start=1 /gene="Bacillus subtilis sacB" /product="secreted levansucrase that renders bacterial growth sensitive to sucrose" /label=SacB /note="negative selection marker" /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV VKDSILEQGQLTVNK" ORIGIN 1 cgggcctaaa gaactaaaaa atctatctgt ttcttttcat tctctgtatt ttttatagtt 61 tctgttgcat gggcataaag ttgcaagctt gatatcttga cagctagctc agtcctaggt 121 ataatactag tcgagaccat tggtctcagt tttagagcta gaaatagcaa gttaaaataa 181 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt tgatatcgaa 241 ttcctgcagc ccgggggatc cactagttct agagcggccg ccaccgcggt ggagctcatc 301 ccaatggcgc gccgagcttg gctcgagcat ggtcatagct gtttcctgtg tgaaattgtt 361 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 421 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 481 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 541 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 601 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 661 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 721 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 781 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 841 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 901 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 961 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 1021 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 1081 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 1141 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 1201 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 1261 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 1321 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 1381 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 1441 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttagaaaa 1501 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt 1561 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg 1621 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt 1681 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg 1741 gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac 1801 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag 1861 cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc 1921 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta 1981 atacctggaa tgctgttttc ccagggatcg cagtggtgag taaccatgca tcatcaggag 2041 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga 2101 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg 2161 gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc 2221 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctagagc 2281 aagacgtttc ccgttgaata tggctcatac tcttcctttt tcaatattat tgaagcattt 2341 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 2401 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta 2461 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg 2521 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt 2581 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc 2641 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt 2701 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca 2761 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg 2821 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac 2881 gacgttgtaa aacgacggcc agtgaattga cgcgtattgg gatggtaccg ggccccccct 2941 cgaggtcgac ggtatcgata cggcatcaga gcagattgta ctgagagtgc accataatcg 3001 gcattttctt ttgcgttttt atttgttaac tgttaattgt ccttgttcaa ggatgctgtc 3061 tttgacaaca gatgttttct tgcctttgat gttcagcagg aagctaggcg caaacgttga 3121 ttgtttgtct gcgtagaatc ctctgtttgt catatagctt gtaatcacga cattgtttcc 3181 tttcgcttga ggtacagcga agtgtgagta agtaaaggtt acatcgttag gatcaagatc 3241 catttttaac acaaggccag ttttgttcag cggcttgtat gggccagtta aagaattaga 3301 aacataacca agcatgtaaa tatcgttaga cgtaatgccg tcaatcgtca tttttgatcc 3361 gcgggagtca gtgaacagat accatttgcc gttcatttta aagacgttcg cgcgttcaat 3421 ttcatctgtt actgtgttag atgcaatcag cggtttcatc acttttttca gtgtgtaatc 3481 atcgtttagc tcaatcatac cgagagcgcc gtttgctaac tcagccgtgc gttttttatc 3541 gctttgcaga agtttttgac tttcttgacg gaagaatgat gtgcttttgc catagtatgc 3601 tttgttaaat aaagattctt cgccttggta gccatcttca gttccagtgt ttgcttcaaa 3661 tactaagtat ttgtggcctt tatcttctac gtagtgagga tctctcagcg tatggttgtc 3721 gcctgagctg tagttgcctt catcgatgaa ctgctgtaca ttttgatacg tttttccgtc 3781 accgtcaaag attgatttat aatcctctac accgttgatg ttcaaagagc tgtctgatgc 3841 tgatacgtta acttgtgcag ttgtcagtgt ttgtttgccg taatgtttac cggagaaatc 3901 agtgtagaat aaacggattt ttccgtcaga tgtaaatgtg gctgaacctg accattcttg 3961 tgtttggtct tttaggatag aatcatttgc atcgaatttg tcgctgtctt taaagacgcg 4021 gccagcgttt ttccagctgt caatagaagt ttcgccgact ttttgataga acatgtaaat 4081 cgatgtgtca tccgcatttt taggatctcc ggctaatgca aagacgatgt ggtagccgtg 4141 atagtttgcg acagtgccgt cagcgttttg taatggccag ctgtcccaaa cgtccaggcc 4201 ttttgcagaa gagatatttt taattgtgga cgaatcgaac tcaggaactt gatatttttc 4261 atttttttgc tgttcaggga tttgcagcat atcatggcgt gtaatatggg aaatgccgta 4321 tgtttcctta tatggctttt ggttcgtttc tttcgcaaac gcttgagttg cgcctcctgc 4381 cagcagtgcg gtagtaaagg ttaatactgt tgcttgtttt gcaaactttt tgatgttcat 4441 cgttcatgtc tcctttttta tgtactgtgt tagcggtctg cttcttccag ccctcctgtt 4501 tgaagatggc aagttagtta cgcacaataa aaaaagacct aaaatatgta aggggtgacg 4561 ccaaagtata cactttgccc tttacacatt ttaggtcttg cctgctttat cagtaacaaa 4621 cccgcgcgat ttacttttcg acctcattct attagactct cgtttggatt gcaactggtc 4681 tattttcctc ttttgtttga tagaaaatca taaaaggatt tgcagacta //
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