| 质粒名称: | pNW55 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 22988 |
| 存储实验室: | Detlef Weigel |
|
载体骨架基本信息 |
|
| 载体名称: | pBluescriptKS |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 2964 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 植物表达，RNAi；植物基因失活 |
| 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: | osa-MIR528 |
| 物种来源: | O. sativa (rice) |
| 插入大小: | 260 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | SmaI（克隆过程中销毁） |
| 3' 克隆位点: | SmaI（克隆过程中销毁） |
| 5' 测序引物 | T7 |
| 3' 测序引物 | T3 |
|
文献引用方法 |
|
| 材料和方法部分: | pNW55 was a gift from Detlef Weigel (Addgene plasmid # 22988 ; http://n2t.net/addgene:22988 ; RRID:Addgene_22988) |
| 参考文献部分: | Highly specific gene silencing by artificial miRNAs in rice. Warthmann N, Chen H, Ossowski S, Weigel D, Herve P. PLoS One. 2008 . 3(3):e1829. 10.1371/journal.pone.0001829 PubMed 18350165 |
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 pNW55 3224 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pNW55
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3224)
AUTHORS Warthmann N, Chen H, Ossowski S, Weigel D, Herve P
TITLE Highly specific gene silencing by artificial miRNAs in rice.
JOURNAL PLoS One. 2008 . 3(3):e1829.
PUBMED 18350165
REFERENCE 2 (bases 1 to 3224)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3224
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin complement(3..458)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(140..161)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 352..371
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 588..610
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 602..619
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 603..619
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 626..645
/label=T7
/note="T7 promoter, forward primer"
promoter 626..644
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind 677..693
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 677..693
/label=pBluescriptSK
/note="For pBluescript vector"
primer_bind complement(988..1004)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(989..1005)
/label=pBluescriptKS
/note="For pBluescript vector"
promoter complement(1033..1051)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(1072..1088)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1072..1088)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1085..1107)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1096..1112
/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)."
promoter complement(1120..1150)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1165..1186
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1303..1320)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1474..2062)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1554..1573)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2233..3093)
/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 2856..2875
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3094..3198)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ggaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcg cgccattcgc cattcaggct
481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tggagctcca
661 ccgcggtggc ggccgctcta gaactagtgg atccccctcg gatcccagca gcagccacag
721 caaaatttgg tttgggatag gtaggtgtta tgttaggtct ggttttttgg ctgtagcagc
781 agcagtggaa ggggcatgca gaggagcagg agattcagtt tgaagctgga cttcactttt
841 gcctctctct cctgtgcttg cctcttccat tcctgctgct aggctgttct gtggaagttt
901 gcagagttta tattatgggt ttaatcgtcc atggcatcag catcagcagc ggtaccgagg
961 gctgcaggaa ttcgatatca agcttatcga taccgtcgac ctcgaggggg ggcccggtac
1021 cagcttttgt tccctttagt gagggttaat ttcgagcttg gcgtaatcat ggtcatagct
1081 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
1141 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
1201 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
1261 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
1321 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
1381 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
1441 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
1501 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
1561 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
1621 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
1681 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
1741 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
1801 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
1861 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
1921 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
1981 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
2041 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
2101 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
2161 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
2221 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
2281 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
2341 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
2401 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
2461 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
2521 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
2581 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
2641 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
2701 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
2761 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
2821 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
2881 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
2941 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
3001 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
3061 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
3121 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
3181 acaaataggg gttccgcgca catttccccg aaaagtgcca cctg
//
