| 质粒名称: | pTNT-B18R-6His |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 58979 |
| 存储实验室: | Steven Dowdy |
|
载体骨架基本信息 |
|
| 载体名称: | pTNT |
| 载体出品公司: | Promega |
| 空载体大小: | 2871 bp |
| 完整质粒大小: | 3948 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称: | B18R,ankyrin repeat protein |
| 物种来源: | Vaccinia Virus |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | T7/Sp6 |
| 标签/融合蛋白: | His（插入件上的C端子） |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | MfeI（克隆期间销毁） |
| 3’克隆位点: | NotI (未破坏) |
| 5’测序引物 | SP6 |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pTNT-B18R-6His was a gift from Steven Dowdy (Addgene plasmid # 58979 ; http://n2t.net/addgene:58979 ; RRID:Addgene_58979) |
| 参考文献部分: | Efficient generation of human iPSCs by a synthetic self-replicative RNA. Yoshioka N, Gros E, Li HR, Kumar S, Deacon DC, Maron C, Muotri AR, Chi NC, Fu XD, Yu BD, Dowdy SF. Cell Stem Cell. 2013 Aug 1;13(2):246-54. doi: 10.1016/j.stem.2013.06.001. 10.1016/j.stem.2013.06.001 PubMed 23910086 |
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 pTNT-B18R-6His 3948 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION expresses His-tagged B18R protein from Western vaccinia virus that
binds to and neutralizes type I IFNs.
ACCESSION .
VERSION .
KEYWORDS pTNT-B18R-6His
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3948)
AUTHORS Yoshioka N, Gros E, Li HR, Kumar S, Deacon DC, Maron C, Muotri AR,
Chi NC, Fu XD, Yu BD, Dowdy SF
TITLE Efficient generation of human iPSCs by a synthetic self-replicative
RNA.
JOURNAL Cell Stem Cell. 2013 Aug 1;13(2):246-54. doi:
10.1016/j.stem.2013.06.001.
PUBMED 23910086
REFERENCE 2 (bases 1 to 3948)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3948
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 16..34
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 40..58
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 40..57
/label=SP6
/note="SP6 promoter, forward primer"
regulatory 94..103
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1177..1194
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
terminator 1238..1285
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 1364..1819
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1451..1470)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1661..1682
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(1886..1905)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2005..2027
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(2065..2083)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 2151..2255
/gene="bla"
/label=AmpR promoter
CDS 2256..3116
/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 complement(2474..2493)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 3287..3875
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3776..3795
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 taaggctaga gtacttaata cgactcacta taggctagca tttaggtgac actatagaat
61 acaagctact tgttcttttt gcactcgaga attgccacca tgggcacgat gaaaatgatg
121 gtacatatat atttcgtatc attattgtta ttgctattcc acagttacgc catagacatc
181 gaaaatgaaa tcacagaatt cttcaataaa atgagagata ctctaccagc taaagactct
241 aaatggttga atccagcatg tatgttcgga ggcacaatga atgatatagc cgctctagga
301 gagccattca gcgcaaagtg tcctcctatt gaagacagtc ttttatcgca cagatataaa
361 gactatgtgg ttaaatggga aaggctagaa aaaaatagac ggcgacaggt ttctaataaa
421 cgtgttaaac atggtgattt atggatagcc aactatacat ctaaattcag taaccgtagg
481 tatttgtgca ccgtaactac aaagaatggt gactgtgttc agggtatagt tagatctcat
541 attagaaaac ctccttcatg cattccaaaa acatatgaac taggtactca tgataagtat
601 ggcatagact tatactgtgg aattctttac gcaaaacatt ataataatat aacttggtat
661 aaagataata aggaaattaa tatcgacgac attaagtatt cacaaacggg aaaggaatta
721 attattcata atccagagtt agaagatagc ggaagatacg actgttacgt tcattacgac
781 gacgttagaa tcaagaatga tatcgtagta tcaagatgta aaatacttac ggttataccg
841 tcacaagacc acaggtttaa actaatacta gatccaaaaa tcaacgtaac gataggagaa
901 cctgccaata taacatgcac tgctgtgtca acgtcattat tgattgacga tgtactgatt
961 gaatgggaaa atccatccgg atggcttata ggattcgatt ttgatgtata ctctgtttta
1021 actagtagag gcggtattac cgaggcgacc ttgtactttg aaaatgttac tgaagaatat
1081 ataggtaata catataaatg tcgtggacac aactattatt ttgaaaaaac ccttacaact
1141 acagtagtat tggagaagct tgcggccggt ggtggccatc atcatcatca tcattgagag
1201 cggccgcaaa aaaaaaaaaa aaaaaaaaaa aaaaaaacta gcataacccc ttggggcctc
1261 taaacgggtc ttgaggggtt ttttggatcc gggctggcgt aatagcgaag aggcccgcac
1321 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc
1381 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
1441 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
1501 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
1561 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
1621 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
1681 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
1741 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
1801 atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt
1861 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
1921 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
1981 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
2041 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
2101 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
2161 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
2221 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
2281 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
2341 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
2401 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
2461 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
2521 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
2581 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
2641 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
2701 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
2761 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
2821 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
2881 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
2941 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
3001 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
3061 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
3121 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
3181 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
3241 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
3301 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
3361 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
3421 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
3481 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
3541 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
3601 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
3661 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
3721 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
3781 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
3841 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
3901 ggttcctggc cttttgctgg ccttttgctc acatggctcg acagatct
//
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