| 质粒名称: | pUC57-Simple-gRNA backbone |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 51306 |
| 存储实验室: | Yonglong Chen |
|
载体骨架基本信息 |
|
| 载体名称: | pUC57-Simple |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 2710 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | CRISPR; 爪蟾表达 |
| 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 | T7 |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | M13R |
| 3' 测序引物 | M13F |
|
文献引用方法 |
|
| 材料和方法部分: | pUC57-Simple-gRNA backbone was a gift from Yonglong Chen (Addgene plasmid # 51306 ; http://n2t.net/addgene:51306 ; RRID:Addgene_51306) |
| 参考文献部分: | Efficient RNA/Cas9-mediated genome editing in Xenopus tropicalis. Guo X, Zhang T, Hu Z, Zhang Y, Shi Z, Wang Q, Cui Y, Wang F, Zhao H, Chen Y. Development. 2014 Jan 8. 10.1242/dev.099853 PubMed 24401372 |
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 pUC57-Simple-gRNA backbone 2845 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION gRNA expression plasmid.
ACCESSION .
VERSION .
KEYWORDS pUC57-Simple-gRNA backbone
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2845)
AUTHORS Guo X, Zhang T, Hu Z, Zhang Y, Shi Z, Wang Q, Cui Y, Wang F, Zhao H,
Chen Y
TITLE Efficient RNA/Cas9-mediated genome editing in Xenopus tropicalis.
JOURNAL Development. 2014 Jan 8.
PUBMED 24401372
REFERENCE 2 (bases 1 to 2845)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..2845
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 55..76
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 91..121
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 129..145
/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 134..156
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 153..169
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 153..169
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 234..252
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_RNA 271..346
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
primer_bind complement(398..415)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(398..414)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(407..429)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(623..642)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 742..764
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(802..820)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 888..992
/gene="bla"
/label=AmpR promoter
CDS 993..1853
/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(1211..1230)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2024..2612
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2513..2532
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2766..2783
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
61 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
121 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
181 caagctcgct tgcacgcctc tgcactcgtc ggtcccggca tccgataccc aattaatacg
241 actcactata ggtcgtcttc gagaagacct gttttagagc tagaaatagc aagttaaaat
301 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt aaagacccaa
361 tatctcgacg cattcgcgaa gtaccgatct ccaattcact ggccgtcgtt ttacaacgtc
421 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
481 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
541 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
601 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
661 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
721 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
781 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
841 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
901 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
961 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
1021 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
1081 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
1141 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
1201 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
1261 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
1321 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
1381 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
1441 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
1501 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
1561 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
1621 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
1681 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
1741 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
1801 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
1861 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
1921 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
1981 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
2041 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
2101 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
2161 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
2221 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
2281 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
2341 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
2401 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
2461 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
2521 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
2581 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
2641 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
2701 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
2761 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
2821 ccgcgcgttg gccgattcat taatg
//
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