| 质粒名称: | pYLsgRNA-OsU6a/LacZ |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 66195 |
| 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: | -- |
| 基因突变: | -- |
| 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-OsU6a/LacZ was a gift from Yao-Guang Liu (Addgene plasmid # 66195 ; http://n2t.net/addgene:66195 ; RRID:Addgene_66195) |
| 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-OsU6a/LacZ 3491 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION cloning of sgRNAs for expression in plants.
ACCESSION .
VERSION .
KEYWORDS pYLsgRNA-OsU6a/LacZ
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3491)
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 3491)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3491
/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(845..862)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(845..861)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1070..1089)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 1189..1211
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(1249..1267)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 1335..1439
/gene="bla"
/label=AmpR promoter
CDS 1440..2300
/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(1658..1677)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2471..3059
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2960..2979
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3213..3230
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 3347..3368
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3383..3413
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3421..3437
/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 3426..3448
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 3445..3461
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3445..3461
/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 ctaatttttt cctgtagttt tcccacaacc attttttacc atccgaatga taggatagga
421 aaaatatcca agtgaacagt attcctataa aattcccgta aaaagcctgc aatccgaatg
481 agccctgaag tctgaactag ccggtcacct gtacaggcta tcgagatgcc atacaagaga
541 cggtagtagg aactaggaag acgatggttg attcgtcagg cgaaatcgtc gtcctgcagt
601 cgcatctatg ggcctggacg gaatagggga aaaagttggc cggataggag ggaaaggccc
661 aggtgcttac gtgcgaggta ggcctgggct ctcagcactt cgattcgttg gcaccggggt
721 aggatgcaat agagagcaac gtttagtacc acctcgctta gctagagcaa actggactgc
781 cttatatgcg cgggtgctgg cttggctgcc gagagacctc tgaagataac atactaagct
841 tggcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
901 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
961 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc
1021 tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct
1081 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
1141 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
1201 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
1261 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
1321 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
1381 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
1441 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
1501 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
1561 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
1621 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
1681 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
1741 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
1801 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
1861 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
1921 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
1981 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
2041 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
2101 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
2161 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
2221 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
2281 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
2341 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
2401 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
2461 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
2521 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
2581 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
2641 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
2701 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
2761 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
2821 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
2881 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
2941 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
3001 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
3061 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
3121 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
3181 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
3241 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
3301 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
3361 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
3421 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gaattcgagc
3481 tcggtacccg g
//
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