| 质粒名称: | pDG02583 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 104129 |
| 存储实验室: | Dirk Görlich |
|
载体骨架基本信息 |
|
| 载体名称: | pQE80 derivative |
| 载体出品公司: | -- |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 6134 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | NEDD8-specific protease 1 (NEDP1),SENP8 |
| 物种来源: | 二穗短柄草 |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: |
14x Histidine tag（主干上的N末端）
Maltose binding protein（主链N端）
Small ubiquitin-like modifier （SUMO）
cleavage site from Brachypodium distachyon（骨干上的N末端） |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pDG02583 was a gift from Dirk Görlich (Addgene plasmid # 104129 ; http://n2t.net/addgene:104129 ; RRID:Addgene_104129) |
| 参考文献部分: | A toolbox of anti–mouse and anti–rabbit IgG secondary nanobodies. Pleiner T, Bates M, Görlich D. J Cell Biol Dec 2017, jcb.201709115 10.1083/jcb.201709115 |
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 pDG02583 6133 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Optimized bacterial expression plasmid of bdNEDP1 protease (Frey and
Goerlich, 2014).
ACCESSION .
VERSION .
KEYWORDS pDG02583
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6133)
AUTHORS Pleiner T, Bates M, Gorlich D
TITLE A toolbox of anti–mouse and anti–rabbit IgG secondary nanobodies
JOURNAL J Cell Biol Dec 2017, jcb.201709115
REFERENCE 2 (bases 1 to 6133)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6133
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..45
/label=T5 promoter
/note="bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator"
protein_bind 21..37
/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)."
protein_bind 53..69
/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 88..99
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 184..1281
/codon_start=1
/gene="malE (mutated)"
/product="maltose binding protein from E. coli"
/label=MBP
/note="This version of the gene does not encode a signal
sequence, so MBP will remain in the cytosol."
/translation="KTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
YAVRTAVINAASGRQTVDEALKDAQT"
regulatory 1138..1147
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
primer_bind 1255..1278
/label=MBP-F
/note="Maltose binding protein, forward primer"
terminator 2369..2463
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
primer_bind complement(2511..2530)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 2522..3337
/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 complement(2595..2614)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
terminator 3556..3642
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(3735..4817)
/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 4779..4798
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(4818..4895)
/gene="lacI (mutant)"
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
primer_bind complement(4979..5001)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 5087..5227
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 5142..5161
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(5242..5259)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5413..6001)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5493..5512)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6092..6110
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 tcataaaaaa tttatttgct ttgtgagcgg ataacaatta taatagattc aattgtgagc
61 ggataacaat ttcacacaga attcattaaa gaggagaaat taaccatgag caagcatcac
121 catcattcag gccatcacca taccggacac caccatcatt caggcagtca tcaccatacc
181 gggaaaactg aagaaggtaa actggtaatc tggattaacg gcgataaagg ctataacggt
241 ctcgctgaag tcggtaagaa attcgagaaa gataccggaa ttaaagtcac cgttgagcat
301 ccggataaac tggaagagaa attcccacag gttgcggcaa ctggcgatgg ccctgacatt
361 atcttctggg cacacgaccg ctttggtggc tacgctcaat ctggcctgtt ggctgaaatc
421 accccggaca aagcgttcca ggacaagctg tatccgttta cctgggatgc cgtacgttac
481 aacggcaagc tgattgctta cccgatcgct gttgaagcgt tatcgctgat ttataacaaa
541 gatctgctgc cgaacccgcc aaaaacctgg gaagagatcc cggcgctgga taaagaactg
601 aaagcgaaag gtaagagcgc gctgatgttc aacctgcaag aaccgtactt cacctggccg
661 ctgattgctg ctgacggggg ttatgcgttc aagtatgaaa acggcaagta cgacattaaa
721 gacgtgggcg tggataacgc tggcgcgaaa gcgggtctga ccttcctggt tgacctgatt
781 aaaaacaaac acatgaatgc agacactgat tactccatcg cagaagctgc ctttaataaa
841 ggcgaaacag cgatgaccat caacggcccg tgggcatggt ccaacatcga caccagcaaa
901 gtgaattatg gtgtaacggt actgccgacc ttcaagggtc aaccatccaa accgttcgtt
961 ggcgtgctga gcgcaggtat taacgccgcc agtccgaaca aagagctggc aaaagagttc
1021 ctcgaaaact atctgctgac tgatgaaggt ctggaagcgg ttaataaaga caaaccgctg
1081 ggtgccgtag cgctgaagtc ttacgaggaa gagttggcga aagatccacg tattgccgcc
1141 accatggaaa acgcccagaa aggtgaaatc atgccgaaca tcccgcagat gtccgctttc
1201 tggtatgccg tgcgtactgc ggtgatcaac gccgccagcg gtcgtcagac tgtcgatgaa
1261 gccctgaaag acgcgcagac taattcgagc tcagctgcgg gtggcgaaga agataagaaa
1321 ccggcaggtg gcgaaggtgg cggtgcccat atcaacctga aagtgaaagg tcaagacggc
1381 aacgaagtct ttttccgcat caaacgttct acccagctga aaaagctgat gaacgcatac
1441 tgtgaccgtc agtctgtaga catgaccgca attgctttcc tgtttgatgg tcgtcgcctg
1501 cgtgcggaac agaccccgga tgaactggag atggaagatg gcgacgaaat cgacgcaatg
1561 cttcatcaga ctggtggcgg tggaatggat gaacgtgtcc tgtcttatgg tgatgtggtg
1621 ctccttcgtt ccgatctggc tattctgcgt ggcccgcact tcctgaacga ccgtattatc
1681 gccttctatc tggcacacct gtccgcttcc ttccacggtg atggcgatct gctgctcctg
1741 ccgccgagca ttccgtacct gctgtccaac ctgccggacc cggaatctgt cgcagaacca
1801 ctgtgtctgg catcccgccg tctggtcctg ctgccggtta acgataaccc ggacgcctcg
1861 gtggcaaatg gcggttctca ctggactctg ctcgtcctgg acgctgcgac tactgacccg
1921 caggctccgc gtttcgtgca tcatgactcc ctgcgcggtt ccgcgaacgc ggctgctgcg
1981 cgtcgcctgg cccgtgctct gaccgctggt ggtgcgccgc tgcgcttcgt tgaggctcct
2041 accccgaccc agcgtaacgg ccacgactgc ggcgtctatg tcctggccgt agctcgtgct
2101 atttgtggtt ggtggcgttc ttctcgtcgc cgcgaaaacc agcagggcgg tggcggcgat
2161 tggtttgcaa ccatgatgga agaagtggat gctgaatccg taggcgctat gcgcgctgaa
2221 ctgctccagc tcattcaccg tctgattcag gacaaggaac aagaagaaga gaaaaagtcc
2281 aaagctggcg tggaggatac ctgcggccaa taactcgtag gctcgacacg aagcatagcg
2341 gccgcataag cttaattagc tgagcttgga ctcctgttga tagatccagt aatgacctca
2401 gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag
2461 aatccaagct agccatgaaa ataaactgtc tgcttacata aacagtaata caaggggtgt
2521 tatgagccat attcaacggg aaacgtcttg ctctaggccg cgattaaatt ccaacatgga
2581 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
2641 ctatcgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
2701 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
2761 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcttggttac tcacgactgc
2821 gatccccggc aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
2881 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
2941 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
3001 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
3061 agaaatgcac aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
3121 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
3181 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
3241 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
3301 attgcagttt catttgatgc tcgatgagtt tttctaagaa ttaattcatg ggcaaatatt
3361 atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg
3421 atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg
3481 gcggggcgta atttttttaa ggcagttatt ggtgccctta aacgcctggg gtaatgactc
3541 tctagcttga ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
3601 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccctc tagattacgt
3661 gcagtcgatg ataagctgtc aaacatgaga attgtgccta atgagtgagc taacttacat
3721 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
3781 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggggccag ggtggttttt
3841 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt
3901 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt
3961 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc
4021 gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg
4081 ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga
4141 aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga
4201 gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc
4261 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta
4321 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat
4381 aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga
4441 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag
4501 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg
4561 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca
4621 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa
4681 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc
4741 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat
4801 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc
4861 ataccgcgaa aggttttgca ccattcgatg gtgtcggaat ttcgggcagc gttgggtcct
4921 ggccacgggt gcgcatgatc tagagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc
4981 tctgacacat gcagctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagca
5041 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc
5101 agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt
5161 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
5221 catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
5281 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
5341 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
5401 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
5461 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
5521 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
5581 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
5641 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
5701 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
5761 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
5821 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
5881 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
5941 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
6001 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
6061 agggattttg gtcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
6121 tcacctcgag aaa
//
