| 質粒名稱: | PU6::klp-12_sgRNA |
|---|---|
| 出品公司: | 愛迪基因 |
| 目錄編號: | 46170 |
| 存儲實驗室: | John Calarco |
|
載體骨架基本信息 |
|
| 載體名稱: | pUC57 |
| 載體出品公司: | -- |
| 載體抗性: | 氨芐青霉素 |
| 空載體大小: | 2641 bp |
| 完整質粒大小: | 3481 bp |
| 載體修飾: | -- |
| 載體類型: | 蠕蟲表達，CRISPR |
| 篩選標記: | -- |
| 高拷貝/低拷貝: | 高拷貝 |
| 生長條件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名稱: | klp-12 targeting sgRNA,klp-12 |
| 物種來源: | Synthetic |
| 插入大小: | 840 bp |
| 基因突變: | -- |
| GenBank編號 | -- |
| 啟動子 | C. elegans U6 snRNA pol III promoter |
| 標簽/融合蛋白: | -- |
| 克隆方法: | 限制性內切酶 |
| 5' 克隆位點: | EcoRI (未破壞) |
| 3' 克隆位點: | HindIII (未破壞) |
| 5' 測序引物 | M13 (-40) forward |
| 3' 測序引物 | M13 (-48) reverse |
|
文獻引用方法 |
|
| 材料和方法部分: | PU6::klp-12_sgRNA was a gift from John Calarco (Addgene plasmid # 46170 ; //n2t.net/addgene:46170 ; RRID:Addgene_46170) |
| 參考文獻部分: | Heritable genome editing in C. elegans via a CRISPR-Cas9 system. Friedland AE, Tzur YB, Esvelt KM, Colaiacovo MP, Church GM, Calarco JA. Nat Methods. 2013 Jun 30. doi: 10.1038/nmeth.2532. 10.1038/nmeth.2532 PubMed 23817069 |
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PU6::klp-12_sgRNA載體質粒基本信息
PU6::klp-12_sgRNA質粒圖譜載體圖譜和PU6::klp-12_sgRNA載體序列質粒序列多克隆位點信息
PU6::klp-12_sgRNA質粒載體簡介
PU6::klp-12_sgRNA質粒序列載體序列
LOCUS PU6::klp-12_sgRNA 3481 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS PU6::klp-12_sgRNA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3481)
AUTHORS Friedland AE, Tzur YB, Esvelt KM, Colaiacovo MP, Church GM, Calarco
JA
TITLE Heritable genome editing in C. elegans via a CRISPR-Cas9 system.
JOURNAL Nat Methods. 2013 Jun 30. doi: 10.1038/nmeth.2532.
PUBMED 23817069
REFERENCE 2 (bases 1 to 3481)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
//www.snapgene.com
FEATURES Location/Qualifiers
source 1..3481
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(29..51)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 151..170
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 364..386
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 378..395
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 379..395
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 402..901
/label=CeU6 promoter
/note="promoter for the C. elegans K09B11.12 U6 snRNA gene
(Friedland et al., 2013)"
misc_RNA 922..997
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
misc_feature 1005..1241
/label=3' CeU6
/note="region downstream of the C. elegans K09B11.12 U6
snRNA gene"
primer_bind complement(1260..1276)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1260..1276)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1273..1295)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1284..1300
/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(1308..1338)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1353..1374
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1491..1508)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1662..2250)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1742..1761)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2421..3281)
/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 3044..3063
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3282..3386)
/gene="bla"
/label=AmpR promoter
primer_bind 3454..3472
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cctccaagaa ctcgtacaaa
421 aatgctctga agtaggtctc gagatcaata ctgggaagac gaaagtcttg cgaaaccgat
481 tcgctgaccc cagtaaagtc tacttcggta gcccttcccc caccacccag ctcgacgacg
541 tcgacgagta catctacctc ggtcgtcaaa tcaacgccca aaacaacttg atgccggaaa
601 tccaccgaag acgtcgagca gcctgggctg cattcaatgg aatcaagaat gccaccgact
661 ccatcaccga caagaagatt cgtgcgaatc tgttcgactc aattgtcctt ccagcgctca
721 cctacgtttc agaagcctgg acattcacca aagctctatc cgaacgagta cgaatcacac
781 atgcctcccc cacaaattat tgatctacat ccccatttgg tatagtgtct tacaggactc
841 tcaatatgtc agctgcatga taaatggtcc ctatataatt gaattgcaaa tctaaatgtt
901 tgatccacaa gttacaattg ggttttagag ctagaaatag caagttaaaa taaggctagt
961 ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt ttttcaataa tattctagat
1021 atcccttttt gaaactaaaa tgaaatactc taaatttttg agttcaatgt ttcaactaat
1081 cttggttaaa ggtaaattcc cggagggacg attttaaagt tgccagagtt gattaaattg
1141 gtccaaaaaa ttttattagg aaactcgaaa aaaaattgat tttttttgac aatattccaa
1201 agaaaattac aaaatctcga cgccaaacat agtcggctgt gaagcttggc gtaatcatgg
1261 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
1321 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
1381 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
1441 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
1501 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
1561 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
1621 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
1681 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
1741 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
1801 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
1861 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
1921 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
1981 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
2041 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
2101 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
2161 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
2221 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
2281 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
2341 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
2401 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
2461 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
2521 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
2581 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
2641 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
2701 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
2761 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
2821 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
2881 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
2941 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
3001 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
3061 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
3121 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
3181 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
3241 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
3301 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
3361 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
3421 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
3481 c
//
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