| 質粒名稱: | pENTR/pSM2(CMV/TO) (w55-1) |
|---|---|
| 出品公司: | Addgene |
| 目錄編號: | 17388 |
| 存儲實驗室: | Eric Campeau, Paul Kaufman |
|
載體骨架基本信息 |
|
| 載體名稱: | pENTR1A |
| 載體出品公司: | -- |
| 載體抗性: | 卡那霉素 |
| 空載體大小: | 3088 dp |
| 完整質粒大小: | -- |
| 載體修飾: | -- |
| 載體類型: | RNAi ; Entry vector |
| 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: | -- |
| 物種來源: | -- |
| 插入大小: | -- |
| 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: | -- |
| 3' 克隆位點: | -- |
| 5' 測序引物 | ENTRforw |
| 3' 測序引物 | ENTRrev |
|
文獻引用方法 |
|
| 材料和方法部分: | pENTR/pSM2(CMV/TO) (w55-1) was a gift from Eric Campeau & Paul Kaufman (Addgene plasmid # 17388 ; //n2t.net/addgene:17388 ; RRID:Addgene_17388) |
| 參考文獻部分: | A versatile viral system for expression and depletion of proteins in mammalian cells. Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO, Campisi J, Yaswen P, Cooper PK, Kaufman PD.. PLoS One. 2009 Aug 6;4(8):e6529. 10.1371/journal.pone.0006529 PubMed 19657394 |
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pENTR/pSM2(CMV/TO) (w55-1)載體質粒基本信息
pENTR/pSM2(CMV/TO) (w55-1)質粒圖譜載體圖譜和pENTR/pSM2(CMV/TO) (w55-1)載體序列質粒序列多克隆位點信息
(w55-1)質粒圖譜.png)
pENTR/pSM2(CMV/TO) (w55-1)質粒載體簡介
pENTR/pSM2(CMV/TO) (w55-1)質粒序列載體序列
LOCUS pENTR/pSM2(CMV/TO) (w55-1) 3088 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Gateway entry vector for miRNA expression under CMV/TO promoter.
ACCESSION .
VERSION .
KEYWORDS pENTR/pSM2(CMV/TO) (w55-1)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3088)
AUTHORS Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO,
Campisi J, Yaswen P, Cooper PK, Kaufman PD.
TITLE A versatile viral system for expression and depletion of proteins in
mammalian cells.
JOURNAL PLoS One. 2009 Aug 6;4(8):e6529.
PUBMED 19657394
REFERENCE 2 (bases 1 to 3088)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
//www.snapgene.com
FEATURES Location/Qualifiers
source 1..3088
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 103..189
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 281..308
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
primer_bind 316..339
/label=pENTR-F
/note="pENTR vectors, forward primer"
protein_bind 358..457
/gene="mutant version of attL"
/label=attL1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
enhancer 480..783
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 784..987
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 938..958
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
protein_bind 989..1007
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 1010..1028
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
ncRNA 1066..1159
/label=5' miR-30a
/note="sequence upstream of the 71-nt precursor of the
human miR-30a microRNA (Zeng et al., 2002)"
/ncRNA_class="miRNA"
ncRNA 1173..1285
/label=3' miR-30a
/note="sequence downstream of the 71-nt precursor of the
human miR-30a microRNA (Zeng et al., 2002)"
/ncRNA_class="miRNA"
protein_bind complement(1317..1416)
/gene="mutant version of attL"
/label=attL2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(1528..1547)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 1539..2348
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind complement(1606..1625)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
rep_origin 2438..3026
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2927..2946
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
421 ttttttataa tgccaacttt gtacaaaaaa gcaggcttta aaggaacctg ggagttccgc
481 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
541 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
601 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
661 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
721 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
781 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
841 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
901 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
961 cggtgggagg tctatataag cagagctctc cctatcagtg atagagatct ccctatcagt
1021 gatagagatc gtcgactagt cgactaggga taacagggta attgtttgaa tgaggcttca
1081 gtactttaca gaatcgttgc ctgcacatct tggaaacact tgctgggatt acttcttcag
1141 gttaacccaa cagaaggctc gagcaaccag aattcaaggg gctactttag gagcaattat
1201 cttgtttact aaaactgaat accttgctat ctctttgata catttttaca aagctgaatt
1261 aaaatggtat aaattaaatc acttttttca attggaagac taatgcgttt atctagaccc
1321 agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga
1381 acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgcag ctctggcccg
1441 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa
1501 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa
1561 cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
1621 gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc
1681 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
1741 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
1801 ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat
1861 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg tccctgcgcc
1921 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
1981 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
2041 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac
2101 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
2161 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
2221 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
2281 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
2341 ttttctaatc agaattggtt aattggttgt aacattattc agattgggcc ccgttccact
2401 gagcgtcaga cccggtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
2461 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
2521 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
2581 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
2641 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
2701 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
2761 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
2821 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
2881 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
2941 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
3001 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
3061 ccttttgctg gccttttgct cacatgtt
//