| 质粒名称: | pEN_TmiRc3 |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 25748 |
| 存储实验室: | Iain Fraser |
|
载体骨架基本信息 |
|
| 载体名称: | pENTR1A-Gent |
| 载体出品公司: | ATCC 10326362 |
| 空载体大小: | 4608 bp |
| 完整质粒大小: | 4608 bp |
| 载体抗性: | Gentamicin, 10 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DB3.1 |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | attL1(未破坏) |
| 3’克隆位点: | attL2 (未破坏) |
| 5’测序引物 | rpCEP-fwd |
| 3’测序引物 | n/a |
|
文献引用方法 |
|
| 材料和方法部分: | pEN_TmiRc3 was a gift from Iain Fraser (Addgene plasmid # 25748 ; http://n2t.net/addgene:25748 ; RRID:Addgene_25748) |
| 参考文献部分: | A single lentiviral vector platform for microRNA-based conditional RNA interference and coordinated transgene expression. Shin KJ, Wall EA, Zavzavadjian JR, Santat LA, Liu J, Hwang JI, Rebres R, Roach T, Seaman W, Simon MI, Fraser ID. Proc Natl Acad Sci U S A. 2006 Sep 12. 103(37):13759-64. 10.1073/pnas.0606179103 PubMed 16945906 |
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 pEN_TmiRc3 4587 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Entry vector for cloning miR30-based shRNA driven by TRE promoter..
ACCESSION .
VERSION .
KEYWORDS pEN_TmiRc3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4587)
AUTHORS Shin KJ, Wall EA, Zavzavadjian JR, Santat LA, Liu J, Hwang JI,
Rebres R, Roach T, Seaman W, Simon MI, Fraser ID
TITLE A single lentiviral vector platform for microRNA-based conditional
RNA interference and coordinated transgene expression.
JOURNAL Proc Natl Acad Sci U S A. 2006 Sep 12. 103(37):13759-64.
PUBMED 16945906
REFERENCE 2 (bases 1 to 4587)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4587
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 25..48
/label=pENTR-F
/note="pENTR vectors, forward primer"
protein_bind 67..166
/gene="mutant version of attL"
/label=attL1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
protein_bind 205..475
/label=tetracycline response element
/bound_moiety="tetracycline repressor TetR"
/note="contains seven copies of the tetracycline operator
tetO"
promoter 508..546
/label=minimal CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 543..567
/label=LNCX
/note="Human CMV promoter, forward primer"
ncRNA 709..830
/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"
CDS 866..1171
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRIVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
primer_bind 1050..1069
/label=ccdB-fwd
/note="ccdB gene, forward primer"
ncRNA 1192..1318
/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(1357..1456)
/gene="mutant version of attL"
/label=attL2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(1568..1587)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
primer_bind complement(1646..1665)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
CDS complement(2239..2772)
/codon_start=1
/gene="aacC1"
/product="gentamycin acetyltransferase"
/label=GmR
/note="confers resistance to gentamycin"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
promoter complement(2961..2989)
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"
rep_origin 3646..4234
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4135..4154
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
terminator 4399..4485
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 4577..17
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
ORIGIN
1 ctgacggatg gcctttttgc gtttctacaa actcttcctg ttagttagtt acttaagctc
61 gggccccaaa taatgatttt attttgactg atagtgacct gttcgttgca acaaattgat
121 aagcaatgct tttttataat gccaactttg tacaaaaaag caggctttaa aggaaccaat
181 tcagtcgact ggatccttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt
241 taccactccc tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg
301 atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt
361 cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta
421 tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt
481 gaaagtcgag ctcggtaccc gggtcgaggt aggcgtgtac ggtgggaggc ctatataagc
541 agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc
601 catagaagac accgggaccg atccagcctc cgcggccccg aactaggcct aggcgtctga
661 tcactagtga ctctagtcct agtcgactag ggataacagg gtaattgttt gaatgaggct
721 tcagtacttt acagaatcgt tgcctgcaca tcttggaaac acttgctggg attacttctt
781 caggttaacc caacagaagg ctcgagaagg tatattgctg ttgacagtga gcgtgctgca
841 ggtaagcttg aggtgtgctt ccagaatgca gtttaaggtt tacacctata aaagagagag
901 ccgttatcgt ctgtttgtgg atgtacagag tgatattatt gacacgcccg ggcgacggat
961 agtgatcccc ctggccagtg cacgtctgct gtcagataaa gtctcccgtg aactttaccc
1021 ggtggtgcat atcggggatg aaagctggcg catgatgacc accgatatgg ccagtgtgcc
1081 ggtctccgtt atcggggaag aagtggctga tctcagccac cgcgaaaatg acatcaaaaa
1141 cgccattaac ctgatgttct ggggaatata gaattccaac ctgctgcttg cctactgcct
1201 cggaattcaa ggggctactt taggagcaat tatcttgttt actaaaactg aataccttgc
1261 tatctctttg atacattttt acaaagctga attaaaatgg tataaattaa atcacttttt
1321 tcaattggaa gactaatgcg ttttcgagat atctagaccc agctttcttg tacaaagttg
1381 gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa
1441 ataaaatcat tatttgccat ccagctgcag ctctggcccg tgtctcaaaa tctctgatgt
1501 tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac
1561 agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc gcgattaaat
1621 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
1681 ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
1741 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
1801 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
1861 ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata tcctgattca
1921 ggtgaaaata ttgttgatgc gctggcagtg tccctgcgcc ggttgcattc gattcctgtt
1981 tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg
2041 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
2101 caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat
2161 ggtgatttct cacttgataa ccttattttt gacgagggga aattaggccg ggaagccgat
2221 ctcggcttga acgaattgtt aggtggcggt acttgggtcg atatcaaagt gcatcacttc
2281 ttcccgtatg cccaactttg tatagagagc cactgcggga tcgtcaccgt aatctgcttg
2341 cacgtagatc acataagcac caagcgcgtt ggcctcatgc ttgaggagat tgatgagcgc
2401 ggtggcaatg ccctgcctcc ggtgctcgcc ggagactgcg agatcataga tatagatctc
2461 actacgcggc tgctcaaacc tgggcagaac gtaagccgcg agagcgccaa caaccgcttc
2521 ttggtcgaag gcagcaagcg cgatgaatgt cttactacgg agcaagttcc cgaggtaatc
2581 ggagtccggc tgatgttggg agtaggtggc tacgtctccg aactcacgac cgaaaagatc
2641 aagagcagcc cgcatggatt tgacttggtc agggccgagc ctacatgtgc gaatgatgcc
2701 catacttgag ccacctaact ttgttttagg gcgactgccc tgctgcgtaa catcgttgct
2761 gctgcgtaac atcgttgctg ctccataaca tcaaacatcg acccacggcg taacgcgctt
2821 gctgcttgga tgcccgaggc atagactgta caaaaaaaca gtcataacaa gccatgaaaa
2881 ccgccactgc gccgttacca ccgctgcgtt cggtcaaggt tctggaccag ttgcgtgagc
2941 gcatacgcta cttgcattac agtttacgaa ccgaacaggc ttatgtcaac tgggttcgtg
3001 ccttcatccg tttccacggt gtgcgtcacc cggcaacctt gggcagcagc gaagtcgagg
3061 catttctgtc ctggctggcg aacgagcgca aggtttcggt ctccacgcat cgtcaggcat
3121 tggcggcctt gctgttcttc tacggcaagg tgctgtgcac ggatctgccc tggcttcagg
3181 agatcggaag acctcggccg tcgcggcgct tgccggtggt gctgaccccg gatgaagtgg
3241 ttcgcatcct cggttttctg gaaggcgagc atcgtttgtt cgcccaggac tctagctata
3301 gttctagtgg ttggctacta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
3361 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
3421 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
3481 tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca
3541 ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttttc
3601 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
3661 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
3721 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
3781 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
3841 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
3901 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
3961 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
4021 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
4081 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
4141 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
4201 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
4261 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
4321 tgtggataac cgtattaccg ctagcatgga tctcggggac gtctaactac taagcgagag
4381 tagggaactg ccaggcatca aataaaacga aaggctcagt cggaagactg ggcctttcgt
4441 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat
4501 ttgaacgttg tgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc
4561 aggcatcaaa ctaagcagaa ggccatc
//
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