| 质粒名称: | pFGET19_Ulp1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 64697 |
| 存储实验室: | Hideo Iwai |
|
载体骨架基本信息 |
|
| 载体名称: | pET-28b(+) |
| 载体出品公司: | Novagen |
| 空载体大小: | 5368 bp |
| 完整质粒大小: | 5979 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | |
| 载体抗性: | 卡那霉素50 μg/mL |
| 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: | ULP1,SUMO protease ULP1,NIB1 |
| 物种来源: | S. cerevisiae (budding yeast); Saccharomyces cerevisiae |
| 插入大小: | -- |
| 基因突变: | Ulp1(403-621) |
| GenBank编号 | YPL020CNIB1 |
| 启动子: | T7 |
| 标签/融合蛋白: | 6xHis（主干上的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: | EcoRI(未破坏) |
| 5’测序引物 | T7 |
| 3’测序引物 | T7 term |
|
文献引用方法 |
|
| 材料和方法部分: | pFGET19_Ulp1 was a gift from Hideo Iwai (Addgene plasmid # 64697 ; http://n2t.net/addgene:64697 ; RRID:Addgene_64697) |
| 参考文献部分: | Tandem SUMO fusion vectors for improving soluble protein expression and purification. Guerrero F, Ciragan A, Iwai H. Protein Expr Purif. 2015 Aug 20. pii: S1046-5928(15)30039-5. doi: 10.1016/j.pep.2015.08.019. 10.1016/j.pep.2015.08.019 PubMed 26297996 |
买家导航
pFGET19Ulp1载体基本信息
pFGET19Ulp1载体图谱质粒图谱和多克隆位点信息
pFGET19Ulp1载体简介
pFGET19Ulp1载体序列
LOCUS pFGET19_Ulp1 5975 bp ds-DNA circular SYN 17-JUN-2022
DEFINITION H6-Ulp1 from pHYRS52 into pET-28b(+).
ACCESSION .
VERSION .
KEYWORDS pFGET19_Ulp1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5975)
AUTHORS Guerrero F, Ciragan A, Iwai H
TITLE Tandem SUMO fusion vectors for improving soluble protein expression
and purification.
JOURNAL Protein Expr Purif. 2015 Aug 20. pii: S1046-5928(15)30039-5. doi:
10.1016/j.pep.2015.08.019.
PUBMED 26297996
REFERENCE 2 (bases 1 to 5975)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jun 17, 2022 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5975
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 1..20
/label=T7
/note="T7 promoter, forward primer"
promoter 1..19
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 20..44
/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)."
RBS 59..81
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 101..118
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 122..139
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
CDS 840..857
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(910..928)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 924..971
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 1008..1463
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1095..1114)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1305..1326
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(1555..2370)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 2278..2297
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 2362..2381
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 2492..3080
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2981..3000
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3234..3251
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 3266..3405
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(3331..3350)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 3491..3513
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(3507..3698)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(4507..5589)
/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 5551..5570
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(5590..5667)
/gene="lacI"
/label=lacI promoter
primer_bind 5873..5892
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
ORIGIN
1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
61 tgtttaactt taagaaggag atatacatat gcggggttct catcatcatc atcatcatgg
121 tctggttccg cgtggatccc ttgttcctga attaaatgaa aaagacgatg accaagtaca
181 aaaagctttg gcatctagag aaaatactca gttaatgaat agagataata tagagataac
241 agtacgtgat tttaagacct tggcaccacg aagatggcta aatgacacta tcattgagtt
301 ttttatgaaa tacattgaaa aatctacccc taatacagtg gcgtttaatt cgtttttcta
361 taccaattta tcagaaaggg gttatcaagg cgtccggagg tggatgaaga gaaagaagac
421 acaaattgat aaacttgata aaatctttac accaataaat ttgaaccaat cccactgggc
481 gttgggcata attgatttaa aaaagaaaac tataggttac gtagattcat tatcgaatgg
541 tccaaatgct atgagtttcg ctatactgac tgacttgcaa aaatatgtta tggaggaaag
601 taagcataca ataggagaag actttgattt gattcattta gattgtccgc agcaaccaaa
661 tggctacgac tgtggaatat atgtttgtat gaatactctc tatggaagtg cagatgcgcc
721 attggatttt gattataaag atgcgattag gatgagaaga tttattgccc atttgatttt
781 aaccgacgct ttaaaataag aattcgagct ccgtcgacaa gcttgcggcc gcactcgagc
841 accaccacca ccaccactga gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
901 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga
961 ggggtttttt gctgaaagga ggaactatat ccggattggc gaatgggacg cgccctgtag
1021 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
1081 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
1141 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
1201 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
1261 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
1321 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
1381 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
1441 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
1501 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta attcttagaa
1561 aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata
1621 tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat
1681 ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa
1741 tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc
1801 cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt
1861 acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg
1921 agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa
1981 ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc
2041 taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg
2101 agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct
2161 gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc
2221 tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc
2281 gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctaga
2341 gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc
2401 agacagtttt attgttcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
2461 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
2521 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
2581 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
2641 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
2701 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
2761 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
2821 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
2881 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
2941 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
3001 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
3061 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
3121 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
3181 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
3241 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
3301 gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag
3361 ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca
3421 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
3481 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
3541 aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt
3601 tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag
3661 cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga
3721 tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt
3781 actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg
3841 atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat
3901 gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg
3961 cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat
4021 gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc
4081 ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac
4141 aggagcacga tcatgcgcac ccgtggggcc gccatgccgg cgataatggc ctgcttctcg
4201 ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg
4261 aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa
4321 atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata
4381 agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
4441 ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg
4501 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
4561 ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac
4621 cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa
4681 gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg
4741 gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac
4801 gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac
4861 cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga
4921 catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata
4981 tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag
5041 cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc
5101 atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg
5161 aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat
5221 gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac
5281 gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt
5341 aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat
5401 cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc
5461 cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac
5521 cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac
5581 tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg
5641 aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct
5701 gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg
5761 gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
5821 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
5881 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
5941 cggcgtagag gatcgagatc tcgatcccgc gaaat
//
