| 质粒名称: | pYLsgRNA-AtU3b |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 66198 |
| 存储实验室: | Yao-Guang Liu |
|
载体骨架基本信息 |
|
| 载体名称: | based on pUC18 |
| 完整质粒大小: | 3167bp |
| 载体类型: | Cloning Vector |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH10B |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | NONE |
|
文献引用方法 |
|
| 材料和方法部分: |
pYLsgRNA-AtU3b was a gift from Yao-Guang Liu (Addgene plasmid # 66198 ; http://n2t.net/addgene:66198 ; RRID:Addgene_66198)
|
| 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 |
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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 3167 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION cloning of sgRNAs for expression in plants.
ACCESSION .
VERSION .
KEYWORDS pYLsgRNA-AtU3b
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3167)
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 3167)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..3167
/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(521..538)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(521..537)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(530..552)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(746..765)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 865..887
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(925..943)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 1011..1115
/gene="bla"
/label=AmpR promoter
CDS 1116..1976
/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(1334..1353)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2147..2735
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2636..2655
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2889..2906
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 3023..3044
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3059..3089
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3097..3113
/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 3102..3124
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 3121..3137
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3121..3137
/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 gatttacttt aaattttttc
181 ttatgcagcc tgtgatggat aactgaatca aacaaatggc gtctgggttt aagaagatct
241 gttttggcta tgttggacga aacaagtgaa cttttaggat caacttcagt ttatatatgg
301 agcttatatc gagcaataag ataagtgggc tttttatgta atttaatggg ctatcgtcca
361 tagattcact aatacccatg cccagtaccc atgtatgcgt ttcatataag ctcctaattt
421 ctcccacatc gctcaaatct aaacaaatct tgttgtatat ataacactga gggagcaaca
481 ttggtcaaga gacctctgaa gataacatac taagcttggc actggccgtc gttttacaac
541 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
601 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
661 gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
721 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
781 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
841 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
901 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
961 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
1021 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
1081 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
1141 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
1201 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
1261 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
1321 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
1381 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
1441 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
1501 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
1561 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
1621 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
1681 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
1741 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
1801 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
1861 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
1921 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
1981 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
2041 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
2101 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
2161 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
2221 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
2281 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
2341 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
2401 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
2461 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
2521 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
2581 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
2641 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
2701 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
2761 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
2821 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
2881 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
2941 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
3001 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
3061 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
3121 caggaaacag ctatgaccat gattacgaat tcgagctcgg tacccgg
//
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