| 质粒名称: | Flag-TRIM24 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 28138 |
| 存储实验室: | Michelle Barton |
|
载体骨架基本信息 |
|
| 载体名称: | pCMX-(Flag)x2 |
| 载体出品公司: | Barton Lab |
| 空载体大小: | 4500 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | TRIM24 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 3200 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | FLAGX2（插入件上的N端子） |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | XbaI(未破坏) |
| 3’克隆位点: | XhoI (未破坏) |
| 5’测序引物 | T7 |
| 3’测序引物 | 5â € ™ -GTCACACACAGAAGTAAGTAAGGTT-3â € |
|
文献引用方法 |
|
| 材料和方法部分: | Flag-TRIM24 was a gift from Michelle Barton (Addgene plasmid # 28138 ; http://n2t.net/addgene:28138 ; RRID:Addgene_28138) |
| 参考文献部分: | Trim24 targets endogenous p53 for degradation. Allton K, Jain AK, Herz HM, Tsai WW, Jung SY, Qin J, Bergmann A, Johnson RL, Barton MC. Proc Natl Acad Sci U S A. 2009 Jul 14. 106(28):11612-6. 10.1073/pnas.0813177106 PubMed 19556538 |
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 Flag-TRIM24 7617 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Flag-TRIM24
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7617)
AUTHORS Allton K, Jain AK, Herz HM, Tsai WW, Jung SY, Qin J, Bergmann A,
Johnson RL, Barton MC
TITLE Trim24 targets endogenous p53 for degradation.
JOURNAL Proc Natl Acad Sci U S A. 2009 Jul 14. 106(28):11612-6.
PUBMED 19556538
REFERENCE 2 (bases 1 to 7617)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7617
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 48..67
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 212..591
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 592..795
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 746..766
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 840..859
/label=T7
/note="T7 promoter, forward primer"
promoter 840..858
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 872..881
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 884..907
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 911..934
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
intron 4246..4311
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 4441..4461
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 4733..4867
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4770..4789)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4824..4843
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 5325..5459
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5387..5406
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(5524..5541)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5695..6283)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5775..5794)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6454..7314)
/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 7077..7096
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7315..7419)
/gene="bla"
/label=AmpR promoter
primer_bind 7487..7505
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(7543..7565)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
ORIGIN
1 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
61 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
121 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
181 gcgggcctct tcgctattac gccagcgcgt tgacattgat tattgactag ttattaatag
241 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
301 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
361 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
421 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
481 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
541 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
601 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
661 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
721 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
781 tatataagca gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt
841 aatacgactc actataggga gacccaagct tgccaccatg gcagactaca aggacgacga
901 tgacaaggca gactacaagg acgacgatga caagggtacc tctagaagat ctaggacaat
961 ggaggtggcg gtggagaagg cggtggcggc ggcggcagcg gcctcggctg cggcctccgg
1021 ggggccctcg gcggcgccga gcggggagaa cgaggccgag agtcggcagg gcccggactc
1081 ggagcgcggc ggcgaggcgg cccggctcaa cctgttggac acttgcgccg tgtgccacca
1141 gaacatccag agccgggcgc ccaagctgct gccctgcctg cactctttct gccagcgctg
1201 cctgcccgcg ccccagcgct acctcatgct gcccgcgccc atgctgggct cggccgagac
1261 cccgccaccc gtccctgccc ccggctcgcc ggtcagcggc tcgtcgccgt tcgccaccca
1321 agttggagtc attcgttgcc cagtttgcag ccaagaatgt gcagagagac acatcataga
1381 taactttttt gtgaaggaca ctactgaggt tcccagcagt acagtagaaa agtcaaatca
1441 ggtatgtaca agctgtgagg acaacgcaga agccaatggg ttttgtgtag agtgtgttga
1501 atggctctgc aagacgtgta tcagagctca tcagagggta aagttcacaa aagaccacac
1561 tgtcagacag aaagaggaag tatctccaga ggcagttggt gtcaccagcc agcgaccagt
1621 gttttgtcct tttcataaaa aggagcagct gaagctgtac tgtgagacat gtgacaaact
1681 gacatgtcga gactgtcagt tgttagaaca taaagagcat agataccaat ttatagaaga
1741 agcttttcag aatcagaaag tgatcataga tacactaatc accaaactga tggaaaaaac
1801 aaaatacata aaattcacag gaaatcagat ccaaaacaga attattgaag taaatcaaaa
1861 tcaaaagcag gtggaacagg atattaaagt tgctatattt acactgatgg tagaaataaa
1921 taaaaaagga aaagctctac tgcatcagtt agagagcctt gcaaaggacc atcgcatgaa
1981 acttatgcaa caacaacagg aagtggctgg actctctaaa caattggagc atgtcatgca
2041 tttttctaaa tgggcagttt ccagtggcag cagtacagca ttactttata gcaaacgact
2101 gattacatac cggttacggc acctccttcg tgcaaggtgt gatgcatccc cagtgaccaa
2161 caacaccatc caatttcact gtgatcctag tttctgggct caaaatatca tcaacttagg
2221 ttctttagta atcgaggata aagagagcca gccacaaatg cctaagcaga atcctgtcgt
2281 ggaacagaat tcacagccac caagtggttt atcatcaaac cagttatcca agttcccaac
2341 acagatcagc ctagctcaat tacggctcca gcatatgcag caacaggtaa tggctcagag
2401 gcaacaggtg caacggaggc cagcacctgt gggtttacca aaccctagaa tgcaggggcc
2461 catccagcaa ccttccatct ctcatcagca accgcctcca cgtttgataa actttcagaa
2521 tcacagcccc aaacccaatg gaccagttct tcctcctcat cctcaacaac tgagatatcc
2581 accaaaccag aacataccac gacaagcaat aaagccaaac cccctacaga tggctttctt
2641 ggctcaacaa gccataaaac agtggcagat cagcagtgga cagggaaccc catcaactac
2701 caacagcaca tcctctactc cttccagccc cacgattact agtgcagcag gatatgatgg
2761 aaaggctttt ggttcaccta tgatcgattt gagctcacca gtgggagggt cttataatct
2821 tccctctctt ccggatattg actgttcaag tactattatg ctggacaata ttgtgaggaa
2881 agatactaat atagatcatg gccagccaag accaccctca aacagaacgg tccagtcacc
2941 aaattcatca gtgccatctc caggccttgc aggacctgtt actatgacta gtgtacaccc
3001 cccaatacgt tcacctagtg cctccagcgt tggaagccga ggaagctctg gctcttccag
3061 caaaccagca ggagctgact ctacacacaa agtcccagtg gtcatgctgg agccaattcg
3121 aataaaacaa gaaaacagtg gaccaccgga aaattatgat ttccctgttg ttatagtgaa
3181 gcaagaatca gatgaagaat ctaggcctca aaatgccaat tatccaagaa gcatactcac
3241 ctccctgctc ttaaatagca gtcagagctc tacttctgag gagactgtgc taagatcaga
3301 tgcccctgat agtacaggag atcaacctgg acttcaccag gacaattcct caaatggaaa
3361 gtctgaatgg ttggatcctt cccagaagtc acctcttcat gttggagaga caaggaaaga
3421 ggatgacccc aatgaggact ggtgtgcagt ttgtcaaaac ggaggggaac tcctctgctg
3481 tgaaaagtgc cccaaagtat tccatctttc ttgtcatgtg cccacattga caaattttcc
3541 aagtggagag tggatttgca ctttctgccg agacttatct aaaccagaag ttgaatatga
3601 ttgtgatgct cccagtcaca actcagaaaa aaagaaaact gaaggccttg ttaagttaac
3661 acctatagat aaaaggaagt gtgagcgcct acttttattt ctttactgcc atgaaatgag
3721 cctggctttt caagaccctg ttcctctaac tgtgcctgat tattacaaaa taattaaaaa
3781 tccaatggat ttgtcaacca tcaagaaaag actacaagaa gattattcca tgtactcaaa
3841 acctgaagat tttgtagctg attttagatt gatctttcaa aactgtgctg aattcaatga
3901 gcctgattca gaagtagcca atgctggtat aaaacttgaa aattattttg aagaacttct
3961 aaagaacctc tatccagaaa aaaggtttcc caaaccagaa ttcaggaatg aatcagaaga
4021 taataaattt agtgatgatt cagatgatga ctttgtacag ccccggaaga aacgcctcaa
4081 aagcattgaa gaacgccagt tgcttaaact cgagcggccg cccgatatca ccctgcaggg
4141 atcctggcca gctagctagg tagctagagg atctttgtga aggaacctta cttctgtggt
4201 gtgacataat tggacaaact acctacagag atttaaagct ctaaggtaaa tataaaattt
4261 ttaagtgtat aatgtgttaa actactgatt ctaattgttt gtgtatttta gattccaacc
4321 tatggaactg atgaatggga gcagtggtgg aatgccttta atgaggaaaa cctgttttgc
4381 tcagaagaaa tgccatctag tgatgatgag gctactgctg actctcaaca ttctactcct
4441 ccaaaaaaga agagaaaggt agaagacccc aaggactttc cttcagaatt gctaagtttt
4501 ttgagtcatg ctgtgtttag taatagaact cttgcttgct ttgctattta caccacaaag
4561 gaaaaagctg cactgctata caagaaaatt atggaaaaat atttgatgta tagtgccttg
4621 actagagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
4681 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
4741 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
4801 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
4861 ctggatcagc ttcagaagat ggcggagggc ctccaacaca gtaattttcc tcccgactct
4921 taaaatagaa aatgtcaagt cagttaagca ggaagtgact aactgaccgc agctggccgt
4981 gcgacatcct cttttaatta gttgctaggc aactgccctc cagagggcag tgtggttttg
5041 caagaggaag caaaaagcct ctccacccag gcctagaatg tttccaccca atcattacta
5101 tgacaacagc tgtttttttt agtattaagc agaggccggg ggcccctggc ctccgcttac
5161 tctggagaaa aagaagagag gcattgtaga ggcttccaga ggcaacttgt caaaacagga
5221 ctgcttctat ttctgtcaca ctgtctggcc ctgtcacaag gtccagcacc tccatacccc
5281 ctttaataag cagtttggga acgggtgcgg gtcttactcc gcccatcccg cccctaactc
5341 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
5401 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggct
5461 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
5521 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
5581 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
5641 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
5701 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
5761 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
5821 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
5881 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
5941 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
6001 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
6061 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
6121 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
6181 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
6241 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
6301 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
6361 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
6421 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
6481 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
6541 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
6601 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
6661 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
6721 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
6781 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
6841 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
6901 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
6961 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
7021 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
7081 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
7141 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
7201 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
7261 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
7321 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
7381 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
7441 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
7501 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
7561 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtca
//
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