| 質粒名稱: | pRS300 |
|---|---|
| 出品公司: | 愛迪基因 |
| 目錄編號: | 22846 |
| 存儲實驗室: | Detlef Weigel |
|
載體骨架基本信息 |
|
| 載體名稱: | pBluescriptSK+ |
| 載體出品公司: | -- |
| 載體抗性: | 氨芐青霉素 |
| 空載體大小: | 2964 bp |
| 完整質粒大小: | -- |
| 載體修飾: | -- |
| 載體類型: | 植物表達，RNAi；植物基因失活 |
| 篩選標記: | -- |
| 高拷貝/低拷貝: | 高拷貝 |
| 生長條件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名稱: | MIR319a |
| 物種來源: | A. thaliana (mustard weed) |
| 插入大小: | 404 bp |
| 基因突變: | -- |
| GenBank編號 | -- |
| 啟動子 | -- |
| 標簽/融合蛋白: | -- |
| 克隆方法: | 限制性內切酶 |
| 5' 克隆位點: | Sma1（克隆過程中銷毀） |
| 3' 克隆位點: | Sma1（克隆過程中銷毀） |
| 5' 測序引物 | T7 |
| 3' 測序引物 | T3 |
|
文獻引用方法 |
|
| 材料和方法部分: | pRS300 was a gift from Detlef Weigel (Addgene plasmid # 22846 ; //n2t.net/addgene:22846 ; RRID:Addgene_22846) |
| 參考文獻部分: | Highly specific gene silencing by artificial microRNAs in Arabidopsis. Schwab R, Ossowski S, Riester M, Warthmann N, Weigel D. Plant Cell. 2006 May . 18(5):1121-33. 10.1105/tpc.105.039834 PubMed 16531494 |
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pRS300載體質粒基本信息
pRS300質粒圖譜載體圖譜和pRS300載體序列質粒序列多克隆位點信息
pRS300質粒載體簡介
pRS300質粒序列載體序列
LOCUS pRS300 3368 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Backbone for expressing plant artificial miRNAs.
ACCESSION .
VERSION .
KEYWORDS pRS300
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3368)
AUTHORS Schwab R, Ossowski S, Riester M, Warthmann N, Weigel D
TITLE Highly specific gene silencing by artificial microRNAs in
Arabidopsis.
JOURNAL Plant Cell. 2006 May . 18(5):1121-33.
PUBMED 16531494
REFERENCE 2 (bases 1 to 3368)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
//www.snapgene.com
FEATURES Location/Qualifiers
source 1..3368
/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 669..685
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind 670..686
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1124..1140)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1124..1140)
/label=pBluescriptSK
/note="For pBluescript vector"
promoter complement(1177..1195)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(1216..1232)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1216..1232)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1229..1251)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1240..1256
/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(1264..1294)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1309..1330
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1447..1464)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1618..2206)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1698..1717)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2377..3237)
/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 3000..3019
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3238..3342)
/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 tgggtaccgg
661 gccccccctc gaggtcgacg gtatcgataa gcttgatatc gaattcctgc agccccaaac
721 acacgctcgg acgcatatta cacatgttca tacacttaat actcgctgtt ttgaattgat
781 gttttaggaa tatatatgta gagagagctt ccttgagtcc attcacaggt cgtgatatga
841 ttcaattagc ttccgactca ttcatccaaa taccgagtcg ccaaaattca aactagactc
901 gttaaatgaa tgaatgatgc ggtagacaaa ttggatcatt gattctcttt gattggactg
961 aagggagctc cctctctctt ttgtattcca attttcttga ttaatctttc ctgcacaaaa
1021 acatgcttga tccactaagt gacatatatg ctgccttcgt atatatagtt ctggtaaaat
1081 taacattttg ggtttatctt tatttaaggc atcgccatgg ggggatccac tagttctaga
1141 gcggccgcca ccgcggtgga gctccagctt ttgttccctt tagtgagggt taatttcgag
1201 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
1261 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
1321 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
1381 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
1441 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
1501 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
1561 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
1621 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
1681 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
1741 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
1801 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
1861 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
1921 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
1981 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
2041 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
2101 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
2161 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
2221 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
2281 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
2341 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
2401 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
2461 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
2521 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
2581 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
2641 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
2701 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
2761 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
2821 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
2881 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
2941 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
3001 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
3061 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
3121 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
3181 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
3241 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
3301 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
3361 gccacctg
//
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