| 质粒名称: | BLADE-182 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 134914 |
| 存储实验室: | Philippe Mangeot, Théophile Ohlmann, Emiliano Ricci |
|
载体骨架基本信息 |
|
| 载体名称: | 未知 |
| 载体出品公司: | Clontech |
| 空载体大小: | 3323 bp |
| 完整质粒大小: | 3323 bp |
| 载体类型: | CRISPR, Synthetic Biology |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | GFP |
| 物种来源: | Synthetic |
| 启动子: | U6 |
| 克隆方法: | 未知 |
| 5’测序引物 | 未知 |
|
文献引用方法 |
|
| 材料和方法部分: | BLADE-182 was a gift from Philippe Mangeot & Théophile Ohlmann & Emiliano Ricci (Addgene plasmid # 134914 ; http://n2t.net/addgene:134914 ; RRID:Addgene_134914) |
| 参考文献部分: | Genome editing in primary cells and in vivo using viral-derived Nanoblades loaded with Cas9-sgRNA ribonucleoproteins. Mangeot PE, Risson V, Fusil F, Marnef A, Laurent E, Blin J, Mournetas V, Massourides E, Sohier TJM, Corbin A, Aube F, Teixeira M, Pinset C, Schaeffer L, Legube G, Cosset FL, Verhoeyen E, Ohlmann T, Ricci EP. Nat Commun. 2019 Jan 3;10(1):45. doi: 10.1038/s41467-018-07845-z. 10.1038/s41467-018-07845-z PubMed 30604748 |
买家导航
BLADE-182载体质粒基本信息
BLADE-182质粒图谱载体图谱和BLADE-182载体序列质粒序列多克隆位点信息
BLADE-182质粒载体简介
BLADE-182质粒序列载体序列
LOCUS BLADE-182 3321 bp DNA circular SYN 14-APR-2025
DEFINITION sgRNA targeting GFP to be used in nanoblade system.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3321)
AUTHORS Mangeot PE, Risson V, Fusil F, Marnef A, Laurent E, Blin J,
Mournetas V, Massourides E, Sohier TJM, Corbin A, Aube F, Teixeira
M, Pinset C, Schaeffer L, Legube G, Cosset FL, Verhoeyen E, Ohlmann
T, Ricci EP
TITLE Genome editing in primary cells and in vivo using viral-derived
Nanoblades loaded with Cas9-sgRNA ribonucleoproteins.
JOURNAL Nat Commun. 2019 Jan 3;10(1):45. doi: 10.1038/s41467-018-07845-z.
PUBMED 30604748
REFERENCE 2 (bases 1 to 3321)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Commun."; date: "2019-01-3"; volume: "10(1):45. doi"; pages: "
10.1038/s41467-018-07845-z"
COMMENT Sequence Label: BLADE-182
FEATURES Location/Qualifiers
source 1..3321
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(30..458)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(140..161)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 352..371
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 585..607
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 599..616
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 600..616
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 623..642
/label=T7
/note="T7 promoter, forward primer"
promoter 623..641
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind 674..690
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 674..690
/label=pBluescriptSK
/note="For pBluescript vector"
promoter 706..954
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind 706..726
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 877..896
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 976..1051
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter complement(1133..1151)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(1172..1188)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1172..1188)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1185..1207)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1196..1212
/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(join(1220..1226,1227..1244,1245..1250))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1265..1286
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1403..1420)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1574..2162)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1654..1673)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(2333..3124,3125..3193))
/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 2956..2975
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3194..3298)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg agctccaccg
661 cggtggcggc cgctctagaa ctagtggatc ccagggggac ccagagaggg cctatttccc
721 atgattcctt catatttgca tatacgatac aaggctgtta gagagataat tagaattaat
781 ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt
841 gggtagtttg cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact
901 tgaaagtatt tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccgcgagg
961 agctgttcac cgggggtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
1021 tcaacttgaa aaagtggcac cgagtcggtg cttttttgaa ttctagatct tgagacaaat
1081 ggcagtattc atccacaatt ctcgaggggg ggcccggtac cagcttttgt tccctttagt
1141 gagggttaat ttcgagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
1201 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
1261 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
1321 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
1381 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
1441 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
1501 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
1561 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
1621 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
1681 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
1741 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
1801 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
1861 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
1921 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
1981 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
2041 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
2101 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
2161 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
2221 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
2281 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
2341 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
2401 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
2461 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
2521 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
2581 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
2641 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
2701 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
2761 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
2821 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
2881 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
2941 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
3001 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
3061 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
3121 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
3181 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
3241 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
3301 catttccccg aaaagtgcca c
//
