| 質粒名稱: | L4440_BioBrick-sgRNA |
|---|---|
| 出品公司: | Addgene |
| 目錄編號: | 177783 |
| 存儲實驗室: | Michael Ristow |
|
載體骨架基本信息 |
|
| 載體名稱: | L4440 (Addgene #1654) |
| 載體出品公司: | Clontech |
| 完整質粒大小: | 2896bp |
| 載體類型: | 細菌表達 |
| 載體抗性: | 氨芐霉素 |
| 生長溫度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷貝數: | 高拷貝 |
|
插入基因信息 |
|
| 插入基因名稱: | 無 |
|
文獻引用方法 |
|
| 材料和方法部分: | L4440_BioBrick-sgRNA was a gift from Michael Ristow (Addgene plasmid # 177783 ; //n2t.net/addgene:177783 ; RRID:Addgene_177783) |
| 參考文獻部分: | Ingestion of single guide RNAs induces gene overexpression and extends lifespan in C. elegans via CRISPR activation. Fischer F, Benner C, Goyala A, Grigolon G, Vitiello D, Wu J, Zarse K, Ewald CY, Ristow M. J Biol Chem. 2022 May 27:102085. doi: 10.1016/j.jbc.2022.102085. 10.1016/j.jbc.2022.102085 PubMed 35636511 |
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L4440_BioBrick-sgRNA載體質粒基本信息
L4440_BioBrick-sgRNA質粒圖譜載體圖譜和L4440_BioBrick-sgRNA載體序列質粒序列多克隆位點信息
L4440_BioBrick-sgRNA質粒載體簡介
L4440_BioBrick-sgRNA質粒序列載體序列
LOCUS L4440_BioBrick-sgRNA 2896 bp DNA circular SYN 14-APR-2025
DEFINITION Overexpression of sgRNAs in E. coli HT115, sgRNA overexpression
scaffold; for ingestion by C. elegans.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2896)
AUTHORS Fischer F, Benner C, Goyala A, Grigolon G, Vitiello D, Wu J, Zarse
K, Ewald CY, Ristow M
TITLE Ingestion of single guide RNAs induces gene overexpression and
extends lifespan in C. elegans via CRISPR activation.
JOURNAL J Biol Chem. 2022 May 27:102085. doi: 10.1016/j.jbc.2022.102085.
PUBMED 35636511
REFERENCE 2 (bases 1 to 2896)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
//gt12365.cn
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Biol
Chem. 2022 May 27:102085. doi: 10.1016/j.jbc.2022.102085."
COMMENT Sequence Label: L4440_BioBrick-sgRNA
FEATURES Location/Qualifiers
source 1..2896
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 217..236
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(455..559)
/gene="bla"
/label=AmpR promoter
rep_origin complement(585..1040)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(722..743)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 934..953
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1167..1189
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 1181..1198
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 1182..1198
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 1208..1229
/label=BioBrick prefix
/note="BioBrick prefix for parts that do not start with
""ATG"""
promoter 1230..1248
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_RNA 1270..1345
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter 1373..1391
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_RNA 1413..1488
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
misc_feature 1516..1536
/label=BioBrick suffix
/note="universal suffix for all parts"
primer_bind complement(1560..1577)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1731..2319)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1811..1830)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(2490..2896,1..385,386..454))
/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"
ORIGIN
1 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
61 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
121 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
181 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
241 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
301 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
361 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
421 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
481 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
541 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa
601 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
661 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
721 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
781 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
841 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
901 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
961 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
1021 tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg
1081 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
1141 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
1201 gcgcgcggaa ttcgcggccg cttctagagt aatacgactc actatagggc cgtcttcgtt
1261 agaagacctg ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact
1321 tgaaaaagtg gcaccgagtc ggtgcttttt ttaggttctg ttaagtaact gataatacga
1381 ctcactatag ggagagacca gtgtaggtct ctgttttaga gctagaaata gcaagttaaa
1441 ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt tttttaggtt
1501 ctgttaagta actgatacta gtagcggccg ctgcagattt cgataagcca ggttgcttcc
1561 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
1621 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
1681 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
1741 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
1801 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
1861 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
1921 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
1981 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
2041 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
2101 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
2161 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
2221 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
2281 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
2341 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
2401 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
2461 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
2521 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
2581 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg ggacccacgc
2641 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
2701 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
2761 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
2821 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
2881 acatgatccc ccatgt
//
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