| 质粒名称: | pSGKP-km |
|---|---|
| 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 |
|
| 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 |
|
| 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 |
|
| 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 |
买家导航
pSGKP-km载体 pSGKPkm质粒基本信息
pSGKP-km质粒图谱 pSGKPkm载体图谱和pSGKP-km载体序列 pSGKPkm质粒序列多克隆位点信息
pSGKP-km质粒 pSGKPkm载体简介
pSGKP-km质粒序列 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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