| 质粒名称: | HLTV-hTRIM21 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 104973 |
| 存储实验室: | Leo James |
|
载体骨架基本信息 |
|
| 载体名称: | HLTV |
| 载体出品公司: | Mark Allen (MRC LMB) |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 3113 bp |
| 完整质粒大小: | 4513 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Tripartite Motif containing 21 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1428 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | T7 |
| 标签/融合蛋白: | His-Lipoyl（插入物上的N端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BamHI/BglII（在克隆过程中被破坏） |
| 3' 克隆位点: | EcoRI(未破坏) |
| 5' 测序引物 | TAATACGACTCACTATAGGG |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | HLTV-hTRIM21 was a gift from Leo James (Addgene plasmid # 104973 ; http://n2t.net/addgene:104973 ; RRID:Addgene_104973) |
| 参考文献部分: | A Method for the Acute and Rapid Degradation of Endogenous Proteins. Clift D, McEwan WA, Labzin LI, Konieczny V, Mogessie B, James LC, Schuh M. Cell. 2017 Nov 13. pii: S0092-8674(17)31255-2. doi: 10.1016/j.cell.2017.10.033. 10.1016/j.cell.2017.10.033 PubMed 29153837 |
买家导航
HLTVhTRIM21载体基本信息
HLTVhTRIM21载体图谱质粒图谱和多克隆位点信息
HLTVhTRIM21载体简介
HLTVhTRIM21载体序列
LOCUS HLTV-hTRIM21 4456 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Purification of His-Lipoyl-hTRIM21 protein from bacteria.
ACCESSION .
VERSION .
KEYWORDS HLTV-hTRIM21
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4456)
AUTHORS Clift D, McEwan WA, Labzin LI, Konieczny V, Mogessie B, James LC,
Schuh M
TITLE A Method for the Acute and Rapid Degradation of Endogenous Proteins.
JOURNAL Cell. 2017 Nov 13. pii: S0092-8674(17)31255-2. doi:
10.1016/j.cell.2017.10.033.
PUBMED 29153837
REFERENCE 2 (bases 1 to 4456)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4456
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 37..56
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 290..307
/label=L4440
/note="L4440 vector, forward primer"
primer_bind 392..411
/label=T7
/note="T7 promoter, forward primer"
promoter 392..410
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
RBS 442..464
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 475..492
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 769..789
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
primer_bind complement(2229..2247)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 2243..2290
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 2387..2842
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2474..2493)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2684..2705
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 2868..2972
/gene="bla"
/label=AmpR promoter
CDS 2973..3833
/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(3191..3210)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4004..136
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
61 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
121 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
181 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
241 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
301 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
361 tcattaatgc aggatctcga tcccgcgaaa ttaatacgac tcactatagg gagaccacaa
421 cggtttccct ctagaaataa ttttgtttaa ctttaagaag gagatataca tatgcaccac
481 caccaccacc acagcggcgc ttttgaattt aagctgccgg acattggcga aggcatccac
541 gaaggtgaaa ttgtcaaatg gtttgtgaaa ccgggcgatg aagtgaacga agacgatgta
601 ttgtgcgaag tgcaaaatga caaggcggtt gtcgaaattc cctccccggt caaagggaaa
661 gtgcttgaaa tcctcgtccc ggagggaaca gtggcaacgg tcgggcaaac gctcatcacg
721 ctcgatgcgc cgggttatga aaacatgacg accggcagcg acaccggcga aaacctgtac
781 ttccagggtg gatctatggc ttcagcagca cgcttgacaa tgatgtggga ggaggtcaca
841 tgccctatct gcctggaccc cttcgtggag cctgtgagca tcgagtgtgg ccacagcttc
901 tgccaggaat gcatctctca ggttgggaaa ggtgggggca gcgtctgtcc tgtgtgccgg
961 cagcgctttc tgctcaagaa tctccggccc aatcgacagc tagccaacat ggtgaacaac
1021 cttaaagaaa tcagccagga ggccagagag ggcacacagg gggaacggtg tgcagtgcat
1081 ggagagagac ttcacctgtt ctgtgagaaa gatgggaagg ccctttgctg ggtatgtgcc
1141 cagtctcgga aacaccgtga ccacgccatg gtccctcttg aggaggctgc acaggagtac
1201 caggagaagc tccaggtggc attaggggaa ctgagaagaa agcaggagtt ggctgagaag
1261 ttggaagtgg aaattgcaat aaagagagca gactggaaga aaacagtgga aacacagaaa
1321 tctaggattc acgcagagtt tgtgcagcaa aaaaacttcc tggttgaaga agaacagagg
1381 cagctgcagg agctggagaa ggatgagagg gagcagctga gaatcctggg ggagaaagag
1441 gccaagctgg cccagcagag ccaggcccta caggagctca tctcagagct agatcgaagg
1501 tgccacagct cagcactgga actgctgcag gaggtgataa ttgtcctgga aaggagtgag
1561 tcctggaacc tgaaggacct ggatattacc tctccagaac tcaggagtgt gtgccatgtg
1621 ccagggctga agaagatgct gaggacatgt gcagtccaca tcactctgga tccagacaca
1681 gccaatccgt ggctgatact ttcagaagat cggagacaag tgaggcttgg agacacccag
1741 cagagcatac ctggaaatga agagagattt gatagttatc ctatggtcct gggtgcccag
1801 cactttcact ctggaaaaca ttactgggag gtagatgtga caggaaagga ggcctgggac
1861 ctgggtgtct gcagagactc tgtgcgcagg aaggggcact ttttgcttag ttccaagagt
1921 ggcttctgga caatttggtt gtggaacaaa caaaaatatg aggctggcac ctacccccag
1981 actcccctcc accttcaggt gcctccatgc caagttggga ttttcctgga ctatgaggct
2041 ggcatggtct ccttctacaa catcactgac catggctccc tcatctactc cttctctgaa
2101 tgtgccttta caggacctct gcggcccttc ttcagtcctg gtttcaatga tggaggaaaa
2161 aacacagccc ctctaaccct ctgtccactg aatattggat cacaaggatc cactgactat
2221 taagaattcc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
2281 gggttttttg ctgaaaggag gaactatatc cggatctggc gtaatagcga agaggcccgc
2341 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc
2401 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
2461 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
2521 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
2581 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
2641 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
2701 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
2761 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
2821 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
2881 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
2941 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
3001 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
3061 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
3121 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
3181 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
3241 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
3301 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
3361 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
3421 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
3481 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
3541 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
3601 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
3661 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
3721 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
3781 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
3841 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
3901 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
3961 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
4021 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
4081 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
4141 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
4201 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
4261 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
4321 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
4381 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
4441 atccggtaag cggcag
//
