| 质粒名称: | pC0043-PspCas13b crRNA backbone |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 103854 |
| 存储实验室: | Feng Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | pUC19 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 2686 bp |
| 完整质粒大小: | 2962 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达,CRISPR |
| 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: | U6-PspCas13b DR-BbsI-BbsI-polyT |
| 物种来源: | -- |
| 插入大小: | 314 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 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: | pC0043-PspCas13b crRNA backbone was a gift from Feng Zhang (Addgene plasmid # 103854 ; http://n2t.net/addgene:103854 ; RRID:Addgene_103854) |
| 参考文献部分: | RNA editing with CRISPR-Cas13. Cox DBT, Gootenberg JS, Abudayyeh OO, Franklin B, Kellner MJ, Joung J, Zhang F. Science. 2017 Oct 25. pii: eaaq0180. doi: 10.1126/science.aaq0180. 10.1126/science.aaq0180 PubMed 29070703 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pC0043-PspCas13b crRNA backbone 2962 bp ds-DNA circular SYN 21-AUG-2019
DEFINITION For cloning of guide RNAs compatible with PspCas13b. Contains a 3'
direct repeat. Clone using BbsI (BpiI). F overhang cacc. R overhang
caac.
ACCESSION .
VERSION .
KEYWORDS pC0043-PspCas13b crRNA backbone
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2962)
AUTHORS Cox DBT, Gootenberg JS, Abudayyeh OO, Franklin B, Kellner MJ, Joung
J, Zhang F
TITLE RNA editing with CRISPR-Cas13.
JOURNAL Science. 2017 Oct 25. pii: eaaq0180. doi: 10.1126/science.aaq0180.
PUBMED 29070703
REFERENCE 2 (bases 1 to 2962)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 21, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..2962
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 259..278
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 512..529
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 646..667
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 682..712
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 720..736
/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)."
primer_bind 744..760
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 744..760
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 774..1014
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 774..794
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 945..964
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
primer_bind complement(1106..1123)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(1106..1122)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1115..1137)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(1331..1350)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 1450..1472
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(1510..1528)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 1596..1700
/gene="bla"
/label=AmpR promoter
CDS 1701..2561
/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 complement(1919..1938)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2732..358
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
61 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
121 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
181 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
241 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
301 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
361 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
421 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
481 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
541 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
601 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
661 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
721 tgtgagcgga taacaatttc acgcaggaaa cagctatgac catgattacg ccagagggcc
781 tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga gagataattg
841 gaattaattt gactgtaaac acaaagatat tagtacaaaa tacgtgacgt agaaagtaat
901 aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat catatgctta
961 ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttgtggaaa ggacgaaaca
1021 ccatgtcttc ctgggacgaa gacaagttgt ggaaggtcca gttttgaggg gctattacaa
1081 cttttttggt accgagctcg aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
1141 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
1201 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
1261 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt
1321 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
1381 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
1441 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
1501 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
1561 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
1621 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
1681 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
1741 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
1801 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
1861 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
1921 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
1981 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
2041 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
2101 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
2161 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
2221 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
2281 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
2341 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
2401 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
2461 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
2521 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
2581 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
2641 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
2701 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
2761 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
2821 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
2881 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
2941 tcgctctgct aatcctgtta cc
//
