| 质粒名称: | pYLsgRNA-AtU3b/LacZ |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 66199 |
| 存储实验室: | Yao-Guang Liu |
|
载体骨架基本信息 |
|
| 载体名称: | based on pUC18 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | 人细胞色素c氧化酶亚基VIII（human cytochrome C oxidase subunit VIII） |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 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: | -- |
| -- | |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pYLsgRNA-AtU3b/LacZ was a gift from Yao-Guang Liu (Addgene plasmid # 66199 ; http://n2t.net/addgene:66199 ; RRID:Addgene_66199) |
| 参考文献部分: | A robust CRISPR/Cas9 system for convenient, high-efficiency multiplex genome editing in monocot and dicot plants. Ma X, Zhang Q, Zhu Q, Liu W, Chen Y, Qiu R, Wang B, Yang Z, Li H, Lin Y, Xie Y, Shen R, Chen S, Wang Z, Chen Y, Guo J, Chen L, Zhao X, Dong Z, Liu YG. Mol Plant. 2015 Apr 24. pii: S1674-2052(15)00204-X. doi: 10.1016/j.molp.2015.04.007. 10.1016/j.molp.2015.04.007 PubMed 25917172 |
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 pYLsgRNA-AtU3b/LacZ 3369 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION cloning of sgRNAs for expression in plants.
ACCESSION .
VERSION .
KEYWORDS pYLsgRNA-AtU3b/LacZ
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3369)
AUTHORS Ma X, Zhang Q, Zhu Q, Liu W, Chen Y, Qiu R, Wang B, Yang Z, Li H,
Lin Y, Xie Y, Shen R, Chen S, Wang Z, Chen Y, Guo J, Chen L, Zhao X,
Dong Z, Liu YG
TITLE A robust CRISPR/Cas9 system for convenient, high-efficiency
multiplex genome editing in monocot and dicot plants.
JOURNAL Mol Plant. 2015 Apr 24. pii: S1674-2052(15)00204-X. doi:
10.1016/j.molp.2015.04.007.
PUBMED 25917172
REFERENCE 2 (bases 1 to 3369)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3369
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_RNA 20..95
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
primer_bind complement(723..740)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(723..739)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(948..967)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 1067..1089
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(1127..1145)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 1213..1317
/gene="bla"
/label=AmpR promoter
CDS 1318..2178
/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(1536..1555)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2349..2937
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2838..2857
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3091..3108
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 3225..3246
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3261..3291
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3299..3315
/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 3304..3326
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 3323..3339
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3323..3339
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
ORIGIN
1 ggatccagcg tgggtctcgg ttttagagct agaaatagca agttaaaata aggctagtcc
61 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttcaagagct tggagtggat
121 ggaattttcc tccgttttac ctgtggaatc ggcagcaaag gacgcgttga cattgtagga
181 ctatattgct ctaataaagg aggcagctat gctggccgtc gttttacaac gtcgtgactg
241 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
301 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
361 ctaatttact ttaaattttt tcttatgcag cctgtgatgg ataactgaat caaacaaatg
421 gcgtctgggt ttaagaagat ctgttttggc tatgttggac gaaacaagtg aacttttagg
481 atcaacttca gtttatatat ggagcttata tcgagcaata agataagtgg gctttttatg
541 taatttaatg ggctatcgtc catagattca ctaataccca tgcccagtac ccatgtatgc
601 gtttcatata agctcctaat ttctcccaca tcgctcaaat ctaaacaaat cttgttgtat
661 atataacact gagggagcaa cattggtcaa gagacctctg aagataacat actaagcttg
721 gcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
781 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
841 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
901 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
961 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
1021 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
1081 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
1141 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
1201 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
1261 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
1321 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
1381 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
1441 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
1501 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
1561 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
1621 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
1681 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
1741 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
1801 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
1861 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
1921 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
1981 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
2041 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
2101 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
2161 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
2221 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
2281 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
2341 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
2401 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
2461 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
2521 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
2581 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
2641 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
2701 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
2761 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
2821 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
2881 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
2941 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
3001 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
3061 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
3121 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
3181 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
3241 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
3301 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacga attcgagctc
3361 ggtacccgg
//
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