| 质粒名称: | pSUMO1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 52258 |
| 存储实验室: | Primo Schaer |
|
载体骨架基本信息 |
|
| 载体名称: | pCDFDuet-1 |
| 载体出品公司: | Novagen |
| 载体抗性: | 链霉素 |
| 空载体大小: | 3781 bp |
| 完整质粒大小: | 7815 bp |
| 载体修饰: | 带有T7终止子的连接子克隆到pCDFDuet1的两个表达单元之间的AflII位点 |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | SUMO-1,Ubl1,SMT3 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 342 bp |
| 基因突变: | C-terminal gly-gly |
| GenBank编号 | NM_003352.4 |
| 启动子 | T7 |
| 标签/融合蛋白: | 6His标签（主干上的N终端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BglII（克隆过程中销毁） |
| 3' 克隆位点: | PstI（未销毁） |
| 5' 测序引物 | AGGAGATATACCATGGGCAGC |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pSUMO1 was a gift from Primo Schaer (Addgene plasmid # 52258 ; http://n2t.net/addgene:52258 ; RRID:Addgene_52258) |
| 参考文献部分: | Versatile Recombinant SUMOylation System for the Production of SUMO-Modified Protein. Weber AR, Schuermann D, Schar P. PLoS One. 2014 Jul 9;9(7):e102157. doi: 10.1371/journal.pone.0102157. eCollection 2014. 10.1371/journal.pone.0102157 PubMed 25007328 |
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 pSUMO1 7815 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Bacterial expression of human 6His-tagged SUMO1, SUMO-E1 and
SUMO-E2.
ACCESSION .
VERSION .
KEYWORDS pSUMO1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7815)
AUTHORS Weber AR, Schuermann D, Schar P
TITLE Versatile Recombinant SUMOylation System for the Production of
SUMO-Modified Protein.
JOURNAL PLoS One. 2014 Jul 9;9(7):e102157. doi:
10.1371/journal.pone.0102157. eCollection 2014.
PUBMED 25007328
REFERENCE 2 (bases 1 to 7815)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7815
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 2..20
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 21..45
/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 101..118
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
promoter 579..597
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 598..622
/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 1154..1198
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/label=S-Tag
/translation="KETAAAKFERQHMDS"
terminator 1250..1297
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter 1382..1400
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 1401..1425
/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)."
terminator 4514..4561
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(4732..5523)
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
promoter complement(5524..5615)
/gene="bla"
/label=AmpR promoter
rep_origin complement(5663..6401)
/direction=LEFT
/label=CloDF13 ori
/note="Plasmids containing the CloDF13 (CDF) origin of
replication can be propagated in E. coli cells that contain
additional plasmids with compatible origins."
CDS complement(6611..7693)
/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 7655..7674
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(7694..7771)
/gene="lacI"
/label=lacI promoter
ORIGIN
1 ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctgt agaaataatt
61 ttgtttaact ttaataagga gatataccat gggcagcagc catcaccatc atcaccacag
121 ccaggatctg gttccgcgta tgtctgacca ggaggccaaa ccttcaactg aggacttggg
181 ggataagaag gaaggtgaat atattaaact caaagtcatt ggacaggata gcagtgagat
241 tcacttcaaa gtgaaaatga caacacatct caagaaactc aaagaatcat actgtcaaag
301 acagggtgtt ccaatgaatt cactcaggtt tctctttgag ggtcagagaa ttgctgataa
361 tcatactcca aaagaactgg gaatggagga agaagatgtg attgaagttt atcaggaaca
421 aacggggggt tagggatccc cgggaattca tcgtgactga ctgacgctgc aggtcgacaa
481 gcttgcggcc gcataatgct taaccccttg gggcctctaa acgggtcttg aggggttttt
541 tgcatgcatc gtattgtaca cggccgcata atcgaaatta atacgactca ctatagggga
601 attgtgagcg gataacaatt ccccatctta gtatattagt taagtataag aaggagatat
661 acatatgtcg gggatcgccc tcagcagact cgcccaggag aggaaagcat ggaggaaaga
721 ccacccattt ggtttcgtgg ctgtcccaac aaaaaatccc gatggcacga tgaacctcat
781 gaactgggag tgcgccattc cgggaaagaa agggactccg tgggaaggag gcttgtttaa
841 actacggatg cttttcaaag atgattatcc atcttcgcca ccaaaatgta aattcgaacc
901 accattattt cacccgaatg tgtacccttc ggggacagtg tgcctgtcca tcttagagga
961 ggacaaggac tggaggccag ccatcacaat caaacagatc ctattaggaa tacaggaact
1021 tctaaatgaa ccaaatatcc aagacccagc tcaagcagag gcctacacga tttactgcca
1081 aaacagagtg gagtacgaga aaagggtccg agcacaagcc aagaagtttg cgccctcata
1141 actcgagtct ggtaaagaaa ccgctgctgc gaaatttgaa cgccagcaca tggactcgtc
1201 tactagcgca gcttaattaa cctaggctgc tgccaccgct gagcaataac tagcataacc
1261 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaacctcagg catttgagaa
1321 gcacggtacc attaagtcga acagaaagta atcgtattgt acacggccgc ataatcgaaa
1381 ttaatacgac tcactatagg ggaattgtga gcggataaca attccccatc ttagtatatt
1441 agttaagtat aagaaggaga tatacatatg gcagatctca attcccgggt cgacatggca
1501 ctgtcgcggg ggctgccccg ggagctggct gaggcggtgg ccgggggccg ggtgctggtg
1561 gtgggggcgg gcggcatcgg ctgcgagctc ctcaagaatc tcgtgctcac cggtttctcc
1621 cacatcgacc tgattgatct ggatactatt gatgtaagca acctcaacag acagtttttg
1681 tttcaaaaga aacatgttgg aagatcaaag gcacaggttg ccaaggaaag tgtactgcag
1741 ttttacccga aagctaatat cgttgcctac catgacagca tcatgaaccc tgactataat
1801 gtggaatttt tccgacagtt tatactggtt atgaatgctt tagataacag agctgcccga
1861 aaccatgtta atagaatgtg cctggcagct gatgttcctc ttattgaaag tggaacagct
1921 gggtatcttg gacaagtaac tactatcaaa aagggtgtga ccgagtgtta tgagtgtcat
1981 cctaagccga cccagagaac ctttcctggc tgtacaattc gtaacacacc ttcagaacct
2041 atacattgca tcgtttgggc aaagtacttg ttcaaccagt tgtttgggga agaagatgct
2101 gatcaagaag tatctcctga cagagctgac cctgaagctg cctgggaacc aacggaagcc
2161 gaagccagag ctagagcatc taatgaagat ggtgacatta aacgtatttc tactaaggaa
2221 tgggctaaat caactggata tgatccagtt aaacttttta ccaagctttt taaagatgac
2281 atcaggtatc tgttgacaat ggacaaacta tggcggaaaa ggaaacctcc agttccgttg
2341 gactgggctg aagtacaaag tcaaggagaa gaaacgaatg catcagatca acagaatgaa
2401 ccccagttag gcctgaaaga ccagcaggtt ctagatgtaa agagctatgc acgtcttttt
2461 tcaaagagca tcgagacttt gagagttcat ttagcagaaa agggggatgg agctgagctc
2521 atatgggata aggatgaccc atctgcaatg gattttgtca cctctgctgc aaacctcagg
2581 atgcatattt tcagtatgaa tatgaagagt agatttgata tcaaatcaat ggcagggaac
2641 attattcctg ctattgctac tactaatgca gtaattgctg ggttgatagt attggaagga
2701 ttgaagattt tatcaggaaa aatagaccag tgcagaacaa tttttttgaa taaacaacca
2761 aacccaagaa agaagcttct tgtgccttgt gcactggatc ctcccaaccc caattgttat
2821 gtatgtgcca gcaagccaga ggtgactgtg cggctgaatg tccataaagt gactgttctc
2881 accttacaag acaagatagt gaaagaaaaa tttgctatgg tagcaccaga tgtccaaatt
2941 gaagatggga aaggaacaat cctaatatct tccgaagagg gagagacgga agctaataat
3001 cacaagaagt tgtcagaatt tggaattaga aatggcagcc ggcttcaagc agatgacttc
3061 ctccaggact atactttatt gatcaacatc cttcatagtg aagacctagg aaaggacgtt
3121 gaatttgaag ttgttggtga tgctccggaa aaagtggggc ccaaacaagc tgaagatgct
3181 gccaaaagca taaccaatgg cagtgatgat ggagctcagc cctccacctc cacagctcaa
3241 gagcaagatg acgttctcat agttgattcg gatgaagaag attcttcaaa taatgccgac
3301 gtcagtgaag aagagagaag ccgcaagagg aaattagatg agaaagagaa tctcagtgca
3361 aagaggtcac gtatagaaca gaaggaagag cttgatgatg tcatagcatt agattgagcg
3421 gccgcattaa aaggaggaat taactatggt ggagaaggag gaggctggcg gcggcattag
3481 cgaggaggag gcggcacagt atgaccggca gatccgcctg tggggactgg aggcccagaa
3541 acggctgcgg gcctctcggg tgcttcttgt cggcttgaaa ggacttgggg ctgaaattgc
3601 caagaatctc atcttggcag gagtgaaagg actgaccatg ctggatcacg aacaggtaac
3661 tccagaagat cccggagctc agttcttgat tcgtactggg tctgttggcc gaaatagggc
3721 tgaagcctct ttggagcgag ctcagaatct caaccccatg gtggatgtga aggtggacac
3781 tgaggatata gagaagaaac cagagtcatt tttcactcaa ttcgatgctg tgtgtctgac
3841 ttgctgctcc agggatgtca tagttaaagt tgaccagatc tgtcacaaaa atagcatcaa
3901 gttctttaca ggagatgttt ttggctacca tggatacaca tttgccaatc taggagagca
3961 tgagtttgta gaggagaaaa ctaaagttgc caaagttagc caaggagtag aagatgggcc
4021 cgacaccaag agagcaaaac ttgattcttc tgagacaacg atggtcaaaa agaaggtggt
4081 cttctgccct gttaaagaag ccctggaggt ggactggagc agtgagaaag caaaggctgc
4141 tctgaagcgc acgacctccg actactttct ccttcaagtg ctcttaaagt tccgtacaga
4201 taaaggaaga gatcccagtt ctgatacata tgaggaagat tctgagttgt tgctccagat
4261 acgaaatgat gtgcttgact cactgggtat tagtcctgac ctgcttcctg aggactttgt
4321 caggtactgc ttctccgaga tggccccagt gtgtgcggtg gttggaggga ttttggcaca
4381 ggaaattgtg aaggccctgt ctcagcggga ccctcctcac aacaacttct tcttcttcga
4441 tggcatgaag gggaatggga ttgtggagtg ccttggcccc aagggctagg ctgctgccac
4501 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
4561 gctgaaacct caggcatttg agaagcacac ggtcacactg cttccggtag tcaataaacc
4621 ggtaaaccag caatagacat aagcggctat ttaacgaccc tgccctgaac cgacgaccgg
4681 gtcatcgtgg ccggatcttg cggcccctcg gcttgaacga attgttagac attatttgcc
4741 gactaccttg gtgatctcgc ctttcacgta gtggacaaat tcttccaact gatctgcgcg
4801 cgaggccaag cgatcttctt cttgtccaag ataagcctgt ctagcttcaa gtatgacggg
4861 ctgatactgg gccggcaggc gctccattgc ccagtcggca gcgacatcct tcggcgcgat
4921 tttgccggtt actgcgctgt accaaatgcg ggacaacgta agcactacat ttcgctcatc
4981 gccagcccag tcgggcggcg agttccatag cgttaaggtt tcatttagcg cctcaaatag
5041 atcctgttca ggaaccggat caaagagttc ctccgccgct ggacctacca aggcaacgct
5101 atgttctctt gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa
5161 gatacctgca agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg
5221 ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg acttctacag cgcggagaat
5281 ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt
5341 tgtttcatca agccttacgg tcaccgtaac cagcaaatca atatcactgt gtggcttcag
5401 gccgccatcc actgcggagc cgtacaaatg tacggccagc aacgtcggtt cgagatggcg
5461 ctcgatgacg ccaactacct ctgatagttg agtcgatact tcggcgatca ccgcttccct
5521 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
5581 atacatattt gaatgtattt agaaaaataa acaaatagct agctcactcg gtcgctacgc
5641 tccgggcgtg agactgcggc gggcgctgcg gacacataca aagttaccca cagattccgt
5701 ggataagcag gggactaaca tgtgaggcaa aacagcaggg ccgcgccggt ggcgtttttc
5761 cataggctcc gccctcctgc cagagttcac ataaacagac gcttttccgg tgcatctgtg
5821 ggagccgtga ggctcaacca tgaatctgac agtacgggcg aaacccgaca ggacttaaag
5881 atccccaccg tttccggcgg gtcgctccct cttgcgctct cctgttccga ccctgccgtt
5941 taccggatac ctgttccgcc tttctccctt acgggaagtg tggcgctttc tcatagctca
6001 cacactggta tctcggctcg gtgtaggtcg ttcgctccaa gctgggctgt aagcaagaac
6061 tccccgttca gcccgactgc tgcgccttat ccggtaactg ttcacttgag tccaacccgg
6121 aaaagcacgg taaaacgcca ctggcagcag ccattggtaa ctgggagttc gcagaggatt
6181 tgtttagcta aacacgcggt tgctcttgaa gtgtgcgcca aagtccggct acactggaag
6241 gacagatttg gttgctgtgc tctgcgaaag ccagttacca cggttaagca gttccccaac
6301 tgacttaacc ttcgatcaaa ccacctcccc aggtggtttt ttcgtttaca gggcaaaaga
6361 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctact gaaccgctct
6421 agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata
6481 agttgtaatt ctcatgttag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
6541 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat
6601 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
6661 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
6721 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
6781 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
6841 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac
6901 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
6961 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
7021 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
7081 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
7141 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
7201 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
7261 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
7321 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
7381 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
7441 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
7501 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
7561 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
7621 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
7681 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
7741 cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac
7801 tcctgcatta ggaaa
//
