| 质粒名称: | pAV-U6+27-Tornado-F30-TAR Variant-1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 129406 |
| 存储实验室: | Samie Jaffrey |
|
载体骨架基本信息 |
|
| 载体名称: | pAV U6+27 |
| 载体出品公司: | Addgene plasmid # 25709 |
| 空载体大小: | -- |
| 完整质粒大小: | 6243 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄青霉素，100μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称: | 圆形F30-TAR变体-1 |
| 物种来源: | Synthetic |
| 插入大小: | 276 bp |
| 基因突变: | MN052909 MN052909 |
| GenBank编号 | JN247409 |
| 启动子: | U6 |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | Sal1(未破坏) |
| 3’克隆位点: | XbaI (未破坏) |
| 5’测序引物 | GACTATCATATGCTTACCGT |
| 3’测序引物 | GAAATTTGTGATGCTATTGC |
|
文献引用方法 |
|
| 材料和方法部分: | pAV-U6+27-Tornado-F30-TAR Variant-1 was a gift from Samie Jaffrey (Addgene plasmid # 129406 ; http://n2t.net/addgene:129406 ; RRID:Addgene_129406) |
| 参考文献部分: | Live imaging of mRNA using RNA-stabilized fluorogenic proteins. Wu J, Zaccara S, Khuperkar D, Kim H, Tanenbaum ME, Jaffrey SR. Nat Methods. 2019 Sep;16(9):862-865. doi: 10.1038/s41592-019-0531-7. Epub 2019 Aug 30. 10.1038/s41592-019-0531-7 PubMed 31471614 |
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 pAV-U6+27-Tornado-F30-TAR Variant-1 6243 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION circular TAR Variant-1.
ACCESSION .
VERSION .
KEYWORDS pAV-U6+27-Tornado-F30-TAR Variant-1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6243)
AUTHORS Wu J, Zaccara S, Khuperkar D, Kim H, Tanenbaum ME, Jaffrey SR
TITLE Live imaging of mRNA using RNA-stabilized fluorogenic proteins.
JOURNAL Nat Methods. 2019 Sep;16(9):862-865. doi: 10.1038/s41592-019-0531-7.
Epub 2019 Aug 30.
PUBMED 31471614
REFERENCE 2 (bases 1 to 6243)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6243
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 3..332
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 183..318
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 245..264
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 689..1483
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(743..762)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 1353..1372
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
intron 1903..1968
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 2098..2118
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 2543..2677
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2775..2792)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2946..3534)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3026..3045)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3705..4565)
/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 4328..4347
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4566..4670)
/gene="bla"
/label=AmpR promoter
rep_origin 4697..5152
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4784..4803)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4994..5015
/label=F1ori-F
/note="F1 origin, forward primer"
repeat_region 5246..5375
/label=AAV2 ITR
polyA_signal 5424..5545
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
promoter complement(5973..6213)
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind complement(6023..6042)
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
primer_bind complement(6193..6213)
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind complement(6241..18)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
1 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
61 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
121 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
181 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
241 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
301 gaggcttttt tggaggccta ggcttttgca aaaagcttca cgctgccgca agcactcagg
361 gcgcaagggc tgctaaagga agcggaacac gtagaaagcc agtccgcaga aacggtgctg
421 accccggatg aatgtcagct actgggctat ctggacaagg gaaaacgcaa gcgcaaagag
481 aaagcaggta gcttgcagtg ggcttacatg gcgatagcta gactgggcgg ttttatggac
541 agcaagcgaa ccggaattgc cagctggggc gccctctggt aaggttggga agccctgcaa
601 agtaaactgg atggctttct tgccgccaag gatctgatgg cgcaggggat caagatctga
661 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
721 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
781 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
841 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
901 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
961 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
1021 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
1081 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
1141 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
1201 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
1261 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
1321 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
1381 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
1441 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
1501 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
1561 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
1621 cgcggggatc tcatgctgga gttcttcgcc caccccgggc tcgatcccct cgcgagttgg
1681 ttcagctgct gcctgaggct ggacgacctc gcggagttct accggcagtg caaatccgtc
1741 ggcatccagg aaaccagcag cggctatccg cgcatccatg cccccgaact gcaggagtgg
1801 ggaggcacga tggccgcttt ggtccggatc tttgtgaagg aaccttactt ctgtggtgtg
1861 acataattgg acaaactacc tacagagatt taaagctcta aggtaaatat aaaattttta
1921 agtgtataat gtgttaaact actgattcta attgtttgtg tattttagat tccaacctat
1981 ggaactgatg aatgggagca gtggtggaat gcctttaatg aggaaaacct gttttgctca
2041 gaagaaatgc catctagtga tgatgaggct actgctgact ctcaacattc tactcctcca
2101 aaaaagaaga gaaaggtaga agaccccaag gactttcctt cagaattgct aagttttttg
2161 agtcatgctg tgtttagtaa tagaactctt gcttgctttg ctatttacac cacaaaggaa
2221 aaagctgcac tgctatacaa gaaaattatg gaaaaatatt ctgtaacctt tataagtagg
2281 cataacagtt ataatcataa catactgttt tttcttactc cacacaggca tagagtgtct
2341 gctattaata actatgctca aaaattgtgt acctttagct ttttaatttg taaaggggtt
2401 aataaggaat atttgatgta tagtgccttg actagagatc ataatcagcc ataccacatt
2461 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
2521 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
2581 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
2641 gtccaaactc atcaatgtat cttatcatgt ctggatcgat ccaggacctc cctaacccta
2701 tgacgtaatt cacgtcacga ctccacccct ccaggaaccc ctagtgatgg agttggccac
2761 tccctctctg cgcgctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
2821 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
2881 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
2941 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
3001 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
3061 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
3121 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
3181 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
3241 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
3301 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
3361 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
3421 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
3481 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
3541 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
3601 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
3661 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
3721 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
3781 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
3841 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
3901 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
3961 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
4021 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
4081 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
4141 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
4201 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
4261 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
4321 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
4381 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
4441 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
4501 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
4561 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
4621 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
4681 cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
4741 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
4801 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
4861 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
4921 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
4981 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
5041 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
5101 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttccattcgc
5161 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
5221 agcagatctg cagatctgtc cctctctgcg cgctcgctcg ctcactgagg ccgcccgggc
5281 aaagcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag
5341 agagggagtg gccaactcca tcactagggg ttccttgtag ttaatgatta acccgccatg
5401 ctacttatct acgtaagaca tgataagata cattgatgag tttggacaaa ccacaactag
5461 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
5521 cattataagc tgcaataaac aagttactag tcctcggctc gagatcgatg cggtcgccat
5581 atggatatcg gtaaccgacg tcaagcttgc atgcctgcag gtcctagtaa aaagcggacc
5641 gaagtccgct ctagagcgtg gactgtaccc tccactttta tccgggcttg ggaccggcat
5701 tggcagtgtt ctacagtcca cgccgaccgc ggccgcttgc catgaatgat ccggtacctc
5761 gggctcggag ctaatgagct cagacgagcc tttgctagcg gatcatcaga gtatgtggga
5821 aacgtggcaa cgcgtcccac atacacatgg caagcggccg cactcggcat ggttaggcgg
5881 tttcccgccc cctcccattt tatccgggct tgggaccggc gagtgcggcc cgtcgactag
5941 tatatgtgct gccgaagcga gcacggtgtt tcgtcctttc cacaagatat ataaagccaa
6001 gaaatcgaaa tactttcaag ttacggtaag catatgatag tccattttaa aacataattt
6061 taaaactgca aactacccaa gaaattatta ctttctacgt cacgtatttt gtactaatat
6121 ctttgtgttt acagtcaaat taattctaat tatctctcta acagccttgt atcgtatatg
6181 caaatatgaa ggaatcatgg gaaataggcc ctcttcctgc ccgaccttgg atccggctgt
6241 gga
//
