| 質粒名稱: | pGEX-4T-2/Rabaptin-5:R5BD |
|---|---|
| 出品公司: | Addgene |
| 目錄編號: | 168808 |
| 存儲實驗室: | Guangpu Li |
|
載體骨架基本信息 |
|
| 載體名稱: | pGEX-4T-2 |
| 載體出品公司: | Pharmacia/GE Healthcare |
| 空載體大小: | 4970 bp |
| 完整質粒大小: | 5335 bp |
| 載體類型: | 細菌表達 |
| 載體抗性: | 氨芐霉素 |
| 生長溫度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷貝數: | 高拷貝 |
|
插入基因信息 |
|
| 插入基因名稱: | Rab5-binding domain (aa739-862) of Rabaptin-5 |
| 物種來源: | H. sapiens (human) |
| 插入大小: | 374bp |
| GenBank編號 | X91141.1 |
| 啟動子: | CMV |
| 標簽/融合蛋白: | GST (N terminal on backbone) |
| 克隆方法: | 限制性內切酶 |
| 5’克隆位點: | EcoRI (not destroyed) |
| 3’克隆位點: | XhoI (not destroyed) |
| 5’測序引物 | CACGTTTGGTGGTGG |
| 3’測序引物 |
ccgaaacgcgcgagg
(Common Sequencing Primers)
|
|
文獻引用方法 |
|
| 材料和方法部分: | pGEX-4T-2/Rabaptin-5:R5BD was a gift from Guangpu Li (Addgene plasmid # 168808 ; //n2t.net/addgene:168808 ; RRID:Addgene_168808) |
| 參考文獻部分: | Determination of Rab5 activity in the cell by effector pull-down assay. Qi Y, Liang Z, Wang Z, Lu G, Li G. Methods Mol Biol. 2015;1298:259-70. doi: 10.1007/978-1-4939-2569-8_22. 10.1007/978-1-4939-2569-8_22 PubMed 25800849 |
買家導航
pGEX-4T-2/Rabaptin-5:R5BD載體質粒基本信息
pGEX-4T-2/Rabaptin-5:R5BD質粒圖譜載體圖譜和pGEX-4T-2/Rabaptin-5:R5BD載體序列質粒序列多克隆位點信息
pGEX-4T-2/Rabaptin-5:R5BD質粒載體簡介
pGEX-4T-2/Rabaptin-5:R5BD質粒序列載體序列
LOCUS pGEX-4T-2/Rabaptin-5:R5BD 5338 bp DNA circular SYN 14-APR-2025
DEFINITION Production of recombinant GST-R5BD (Rab5-binding domain) in bacteria
for GST pulldown assays to identify binding partners for Rab5
subfamily members.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5338)
AUTHORS Qi Y, Liang Z, Wang Z, Lu G, Li G
TITLE Determination of Rab5 activity in the cell by effector pull-down
assay.
JOURNAL Methods Mol Biol. 2015;1298:259-70. doi:
10.1007/978-1-4939-2569-8_22.
PUBMED 25800849
REFERENCE 2 (bases 1 to 5338)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
//gt12365.cn
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Methods Mol
Biol."; date: "2015"; volume: "1298:259-70. doi"; pages: "
10.1007/978-1-4939-2569-8_22"
COMMENT Sequence Label: pGEX-4T-2/Rabaptin-5:R5BD
FEATURES Location/Qualifiers
source 1..5338
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter join(1..6,7..22,23..29)
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 37..53
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
CDS 76..729
/codon_start=1
/product="glutathione S-transferase from Schistosoma
japonicum"
/label=GST
/translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
primer_bind 687..709
/label=pGEX 5'
/note="pGEX vectors, Glutathione-S-transferase, forward
primer"
CDS 736..753
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
primer_bind complement(1206..1228)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(1373..1391)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 1459..1563
/gene="bla"
/label=AmpR promoter
CDS join(1564..1632,1633..2424)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(1782..1801)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2595..3183
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3084..3103
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3337..3354
/label=L4440
/note="L4440 vector, forward primer"
promoter 3427..3504
/gene="lacI"
/label=lacI promoter
CDS 3505..4587
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=lacI
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind complement(3524..3543)
/label=LacI-R
/note="LacI, reverse primer"
promoter join(4636..4641,4642..4659,4660..4666)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4674..4690
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 4698..4714
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 4698..4714
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
CDS 4710..5237
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
QPSEAVVWLCRS"
primer_bind complement(4730..4747)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(4730..4746)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4739..4761)
/label=M13/pUC Forward
/note="In lacZ gene"
ORIGIN
1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
61 caggaaacag tattcatgtc ccctatacta ggttattgga aaattaaggg ccttgtgcaa
121 cccactcgac ttcttttgga atatcttgaa gaaaaatatg aagagcattt gtatgagcgc
181 gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg gtttggagtt tcccaatctt
241 ccttattata ttgatggtga tgttaaatta acacagtcta tggccatcat acgttatata
301 gctgacaagc acaacatgtt gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt
361 gaaggagcgg ttttggatat tagatacggt gtttcgagaa ttgcatatag taaagacttt
421 gaaactctca aagttgattt tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat
481 cgtttatgtc ataaaacata tttaaatggt gatcatgtaa cccatcctga cttcatgttg
541 tatgacgctc ttgatgttgt tttatacatg gacccaatgt gcctggatgc gttcccaaaa
601 ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa ttgataagta cttgaaatcc
661 agcaagtata tagcatggcc tttgcagggc tggcaagcca cgtttggtgg tggcgaccat
721 cctccaaaat cggatctggt tccgcgtgga tccccaggaa ttcgagcttc tatttctagc
781 ctaaaagctg aattagaaag aataaaagtg gaaaaaggac agttggagtc cacattaaga
841 gagaagtctc aacagcttga gagtcttcag gaaataaaga tcagtttgga agagcagtta
901 aagaaagaga ctgctgctaa ggctaccgtt gaacagctaa tgtttgaaga gaagaacaaa
961 gctcagagat tacagacaga attagatgtc agtgagcaag tccagagaga ttttgtaaag
1021 ctttcacaga cccttcaggt gcagttagag cggatccggc aagctgactc cttggagaga
1081 atccgggcaa ttctgaatga tactaaactg acagacatta accagcttcc tgagacatga
1141 ctcgagcggc cgcatcgtga ctgactgacg atctgcctcg cgcgtttcgg tgatgacggt
1201 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
1261 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc
1321 atgacccagt cacgtagcga tagcggagtg tataattctt gaagacgaaa gggcctcgtg
1381 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
1441 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
1501 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
1561 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
1621 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
1681 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
1741 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
1801 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
1861 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
1921 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
1981 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
2041 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
2101 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
2161 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
2221 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
2281 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
2341 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
2401 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
2461 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
2521 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
2581 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
2641 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
2701 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
2761 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
2821 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
2881 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
2941 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
3001 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
3061 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
3121 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
3181 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
3241 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
3301 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
3361 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
3421 aattccgaca ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa
3481 gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat
3541 gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg
3601 aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg
3661 gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc
3721 ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc
3781 agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac
3841 aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat
3901 gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac
3961 cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag
4021 catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc
4081 tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg
4141 atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg
4201 ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc
4261 gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga
4321 tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat
4381 tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg
4441 gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc
4501 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
4561 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt agctcactca
4621 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
4681 cggataacaa tttcacacag gaaacagcta tgaccatgat tacggattca ctggccgtcg
4741 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
4801 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
4861 agttgcgcag cctgaatggc gaatggcgct ttgcctggtt tccggcacca gaagcggtgc
4921 cggaaagctg gctggagtgc gatcttcctg aggccgatac tgtcgtcgtc ccctcaaact
4981 ggcagatgca cggttacgat gcgcccatct acaccaacgt gacctatccc attacggtca
5041 atccgccgtt tgttcccacg gagaatccga cgggttgtta ctcgctcaca tttaatgttg
5101 atgaaagctg gctacaggaa ggccagacgc gaattatttt tgatggcgtt ggaattagct
5161 tatcgactgc acggtgcacc aatgcttctg gcgtcaggca gccatcggaa gctgtggtat
5221 ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct caaggcgcac tcccgttctg
5281 gataatgttt tttgcgccga catcataacg gttctggcaa atattctgaa atgagctg
//