| 质粒名称: | pXR004: CasRx pre-gRNA cloning backbone |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 109054 |
| 存储实验室: | Patrick Hsu |
|
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: | -- |
| 载体修饰: | 插入gRNA表达盒 |
| 载体类型: | 哺乳动物表达,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: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | hU6 |
| 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: | pXR004: CasRx pre-gRNA cloning backbone was a gift from Patrick Hsu (Addgene plasmid # 109054 ; http://n2t.net/addgene:109054 ; RRID:Addgene_109054) |
| 参考文献部分: | Transcriptome Engineering with RNA-Targeting Type VI-D CRISPR Effectors. Konermann S, Lotfy P, Brideau NJ, Oki J, Shokhirev MN, Hsu PD. Cell. 2018 Mar 8. pii: S0092-8674(18)30207-1. doi: 10.1016/j.cell.2018.02.033. 10.1016/j.cell.2018.02.033 PubMed 29551272 |
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 pXR004: CasRx pre-gRNA cloning backbone 2945 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION hU6-driven expression of guide RNAs compatible with CasRx. Contains
BbsI sites for guide cloning flanked by 5' and 3' full-length DRs.
ACCESSION .
VERSION .
KEYWORDS pXR004: CasRx pre-gRNA cloning backbone
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2945)
AUTHORS Konermann S, Lotfy P, Brideau NJ, Oki J, Shokhirev MN, Hsu PD
TITLE Transcriptome Engineering with RNA-Targeting Type VI-D CRISPR
Effectors.
JOURNAL Cell. 2018 Mar 8. pii: S0092-8674(18)30207-1. doi:
10.1016/j.cell.2018.02.033.
PUBMED 29551272
REFERENCE 2 (bases 1 to 2945)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..2945
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 7..247
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 7..27
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 178..197
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
rep_origin 428..883
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(515..534)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 725..746
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(900..919)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 1019..1041
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(1079..1097)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 1165..1269
/gene="bla"
/label=AmpR promoter
CDS 1270..2130
/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(1488..1507)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2301..2889
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2790..2809
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 gctagcgagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
61 agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
121 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
181 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
241 aaaggacgaa acaccgcaag taaaccccta ccaactggtc ggggtttgaa acgggtcttc
301 gagaagacct caagtaaacc cctaccaact ggtcggggtt tgaaactttt tttgaattct
361 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
421 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
481 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
541 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
601 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
661 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
721 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
781 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
841 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
901 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
961 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
1021 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
1081 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
1141 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
1201 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
1261 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
1321 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
1381 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
1441 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
1501 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
1561 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
1621 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
1681 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
1741 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
1801 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
1861 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
1921 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
1981 cgtggaagcc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
2041 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
2101 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
2161 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
2221 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
2281 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
2341 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
2401 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
2461 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
2521 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
2581 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
2641 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
2701 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
2761 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
2821 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
2881 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
2941 gctca
//
