| 质粒名称: | pDisCoTune |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 173713 |
| 存储实验室: | Morten Norholm |
|
载体骨架基本信息 |
|
| 载体名称: | pDisCoTune |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 载体抗性: | 氯霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | ERV1 |
| 物种来源: | S. cerevisiae (budding yeast) |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5’克隆位点: | -- |
| 3’克隆位点: | --- |
| 5’测序引物 | gccagcccaaatgcaatc |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pDisCoTune was a gift from Morten Norholm (Addgene plasmid # 173713 ; http://n2t.net/addgene:173713 ; RRID:Addgene_173713) |
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 pDisCoTune 7358 bp ds-DNA circular SYN 16-SEP-2021
DEFINITION Promotes expression of disulfide-rich protein in E. coli T7
expression system. The plasmid encodes expression of the folding
factors Erv1p and hPDI controlled by the Ptac promoter..
ACCESSION .
VERSION .
KEYWORDS pDisCoTune
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7358)
TITLE DisCoTune: versatile auxiliary plasmids for production of
disulfide-containing proteins and peptides in the E. coli T7 system
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 7358)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 16, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7358
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 85..540
/codon_start=1
/product="lysozyme from bacteriophage T7"
/label=T7 lysozyme
/note="inhibitor of T7 RNA polymerase"
/translation="MARVQFKQRESTDAIFVHCSATKPSQNVGVREIRQWHKEQGWLDV
GYHFIIKRDGTVEAGRDEMAVGSHAKGYNHNSIGVCLVGGIDDKGKFDANFTPAQMQSL
RSLLVTLLAKYEGAVLRAHHEVAPYACPSFDLKRWWEKNELVTSDRG"
terminator 610..657
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter 693..721
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 729..745
/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 734..756
/label=M13/pUC Reverse
/note="In lacZ gene"
RBS 1431..1453
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 2993..3010
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS complement(3233..3892)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind 3807..3826
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(3893..3995)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
primer_bind complement(4387..4404)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4521..5066)
/direction=LEFT
/label=p15A ori
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin.
"
primer_bind complement(4601..4620)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5397..6245)
/codon_start=1
/gene="E. coli rhaR"
/product="transcriptional activator of rhaR and rhaS"
/label=rhaR
/note="needed for rhamnose-dependent activation of the rhaB
promoter"
/translation="MAHQLKLLKDDFFASDQQAVAVADRYPQDVFAEHTHDFCELVIVW
RGNGLHVLNDRPYRITRGDLFYIHADDKHSYASVNDLVLQNIIYCPERLKLNLDWQGAI
PGFNASAGQPHWRLGSMGMAQARQVIGQLEHESSQHVPFANEMAELLFGQLVMLLNRHR
YTSDSLPPTSSETLLDKLITRLAASLKSPFALDKFCDEASCSERVLRQQFRQQTGMTIN
QYLRQVRVCHAQYLLQHSRLLISDISTECGFEDSNYFSVVFTRETGMTPSQWRHLNSQK
D"
CDS complement(6319..7155)
/codon_start=1
/gene="E. coli rhaS"
/product="positive regulator of the rhaB promoter
"
/label=rhaS
/note="activated by L-rhamnose"
/translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
GIHVFNGQPYTITCGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
promoter 7301..61
/label=rhaB promoter
/note="promoter of the E. coli rhaBAD operon, conferring
tight induction with L-rhamnose and repression with
D-glucose in the presence of RhaR and RhaS (Giacalone et
al., 2006)"
ORIGIN
1 gcccattttc ctgtcagtaa cgagaaggtc gcgaattcag gcgcttttta gactggtcgt
61 aatgaaattc aggaggtggt cgacatggct cgtgtacagt ttaaacaacg tgaatctact
121 gacgcaatct ttgttcactg ctcggctacc aagccaagtc agaatgttgg tgtccgtgag
181 attcgccagt ggcacaaaga gcagggttgg ctcgatgtgg gataccactt tatcatcaag
241 cgagacggta ctgtggaggc aggacgagat gagatggctg taggctctca cgctaagggt
301 tacaaccaca actctatcgg cgtctgcctt gttggtggta tcgacgataa aggtaagttc
361 gacgctaact ttacgccagc ccaaatgcaa tcccttcgct cactgcttgt cacactgctg
421 gctaagtacg aaggcgctgt gcttcgcgcc catcatgagg tggcgccgta cgcttgccct
481 tcgttcgacc ttaagcgttg gtgggagaag aacgaactgg tcacttctga ccgtggataa
541 ggatccccgc gccctcatcc gaaagggcgt attcatatgc cctaggctgc tgccaccgct
601 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgcat
661 aacggttctg gcaaatattc tgaaatgagc tgttgacaat taatcatcgg ctcgtataat
721 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agccagtccg tttaggtgtt
781 ttcacgagct ctagaaataa ttatcagcag gacgcactga ccaggaggta catatgaaag
841 ccatcgataa aatgaccgac aatccgcctc aagagggtct gagtggtcgt aaaatcatct
901 atgatgagga cggcaaacca tgtcgcagct gcaataccct gctggacttc cagtatgtta
961 ccgggaaaat ctcaaatggc ctgaaaaacc tgagcagcaa tggtaaactg gcgggtacag
1021 gtgctctgac tggggaagca tctgaactga tgccgggttc tcgtacctat cgtaaagtcg
1081 atcctccgga tgttgaacag ctgggacgtt cttcatggac actgctgcat agcgtagcag
1141 cctcttatcc agctcaacca accgatcagc aaaaaggcga aatgaaacag ttcctgaaca
1201 tcttcagcca catctatccg tgtaactggt gcgctaaaga cttcgaaaaa tatatccgtg
1261 agaatgcccc tcaagttgaa tcacgtgagg agctgggtcg ctggatgtgt gaggcccata
1321 acaaagtgaa caaaaagctg cgcaaaccga agttcgactg taacttctgg gaaaaacgct
1381 ggaaagatgg ttgggatgag taataaggat ccgaattcac tagaaataat tttgtttaac
1441 tttaagaagg agatacatat ggatgcccca gaggaggaag atcacgtcct ggttctgcgt
1501 aaaagcaact tcgctgaagc actggcagct cacaaatatc tgctggtcga gttctatgct
1561 ccgtggtgtg gtcattgcaa agccctggct ccggaatatg ctaaagcagc cggcaaactg
1621 aaagctgagg gcagtgaaat tcgtctggcc aaagtggacg ctaccgaaga atcagatctg
1681 gcacaacagt atggtgttcg tggttatccg actatcaaat ttttccgtaa cggcgataca
1741 gcaagcccta aagagtatac cgctggccgt gaagctgatg atatcgtgaa ctggctgaaa
1801 aaacgtacag gtccggcggc aacgactctg cctgatggtg ccgctgccga gtcactggta
1861 gaatcatccg aagtggccgt gattggcttc tttaaagacg tggagagcga ttcagcaaaa
1921 cagttcctgc aagcagctga agcgattgat gacatcccgt ttggtattac gagcaatagc
1981 gacgtgttct ccaaatatca actggacaaa gacggtgtgg ttctgttcaa aaaattcgac
2041 gaaggccgta acaactttga aggtgaggtg accaaagaaa acctgctgga ctttatcaaa
2101 cacaatcaac tgccgctggt gattgagttc accgaacaga cagctccgaa aatctttggc
2161 ggcgagatca aaacccacat tctgctgttt ctgcctaaaa gtgtgtccga ctatgacggc
2221 aaactgagca acttcaaaac cgccgctgag agctttaaag gcaaaatcct gttcatcttc
2281 atcgacagcg atcataccga caaccagcgt attctggagt tttttggcct gaaaaaagag
2341 gaatgtccgg ccgttcgcct gattacactg gaagaggaga tgacgaaata taaaccagaa
2401 agcgaggagc tgacagctga acgtattacc gagttctgcc accgttttct ggaaggcaaa
2461 atcaaacctc atctgatgtc ccaagaactg ccagaagatt gggataaaca gcctgtgaaa
2521 gtgctggtag gcaaaaactt cgaggatgtg gcgttcgacg agaaaaaaaa cgtgtttgtg
2581 gagttttatg ccccttggtg tggacactgc aaacagctgg caccgatttg ggataaactg
2641 ggcgaaacct ataaagatca tgaaaacatt gttattgcca aaatggacag caccgccaat
2701 gaagtcgaag ccgtgaaagt tcattcgttt ccgaccctga aattctttcc tgccagcgcc
2761 gatcgtactg tgatcgatta taacggggag cgtaccctgg atggttttaa aaaattcctg
2821 gagagcggtg gtcaagatgg tgccggtgat gatgacgatc tggaggatct ggaagaggct
2881 gaagaaccgg atatggagga ggatgacgat cagaaagcag tcaaagacga gctgtgataa
2941 ggatccgaat tcactagtga gctccgtcga caagcttgcg gccgcactcg agcaccacca
3001 ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
3061 caccgctgag caataactag cagataaaaa aaatccttag ctttcgctaa ggatgatttc
3121 tactggttat ggtcgaattt gctttcgaat ttctgccatt catccgctta ttatcactta
3181 ttcaggcgta gcaaccaggc gtttaagggc accaataact gccttaaaaa aattacgccc
3241 cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg acatggaagc
3301 catcacaaac ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg
3361 tataatattt gcccatagtg aaaacggggg cgaagaagtt gtccatattg gccacgttta
3421 aatcaaaact ggtgaaactc acccagggat tggctgagac gaaaaacata ttctcaataa
3481 accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt
3541 gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt
3601 gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt
3661 tcattgccat acggaactcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg
3721 ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagct
3781 gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac
3841 gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt
3901 ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat
3961 tatggtgaaa gttggaacct cttacgtgcc gatcaacgtc tcattttcgc caaaagttgg
4021 cccagggctt cccggtatca acagggacac caggatttat ttattctgcg aagtgatctt
4081 ccgtcacagg tatttattcg gcgcaaagtg cgtcgggtga tgctgccaac ttactgattt
4141 agtgtatgat ggtgtttttg aggtgctcca gtggcttctg tttctatcag ctgtccctcc
4201 tgttcagcta ctgacggggt ggtgcgtaac ggcaaaagca ccgccggaca tcagcgctag
4261 cggagtgtat actggcttac tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg
4321 tggcaggaga aaaaaggctg caccggtgcg tcagcagaat atgtgataca ggatatattc
4381 cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc
4441 ttacgaacgg ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag
4501 agggccgcgg caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc
4561 tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
4621 cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg
4681 ctgttatggc cgcgtttgtc tcattccacg cctgacactc agttccgggt aggcagttcg
4741 ctccaagctg gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg
4801 taactatcgt cttgagtcca acccggaaag acatgcaaaa gcaccactgg cagcagccac
4861 tggtaattga tttagaggag ttagtcttga agtcatgcgc cggttaaggc taaactgaaa
4921 ggacaagttt tggtgactgc gctcctccaa gccagttacc tcggttcaaa gagttggtag
4981 ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca gagcaagaga
5041 ttacgcgcag accaaaacga tctcaagaag atcatcttat taatcagata aaatatttct
5101 aggtttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata
5161 agttgtaatt ctcatgttag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
5221 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat
5281 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
5341 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgcccgataa gcttaattaa
5401 tctttctgcg aattgagatg acgccactgg ctgggcgtca tcccggtttc ccgggtaaac
5461 accaccgaaa aatagttact atcttcaaag ccacattcgg tcgaaatatc actgattaac
5521 aggcggctat gctggagaag atattgcgca tgacacactc tgacctgtcg cagatattga
5581 ttgatggtca ttccagtctg ctggcgaaat tgctgacgca aaacgcgctc actgcacgat
5641 gcctcatcac aaaatttatc cagcgcaaag ggacttttca ggctagccgc cagccgggta
5701 atcagcttat ccagcaacgt ttcgctggat gttggcggca acgaatcact ggtgtaacga
5761 tggcgattca gcaacatcac caactgcccg aacagcaact cagccatttc gttagcaaac
5821 ggcacatgct gactactttc atgctcaagc tgaccaataa cctgccgcgc ctgcgccatc
5881 cccatgctac ctaagcgcca gtgtggttgc cctgcgctgg cgttaaatcc cggaatcgcc
5941 ccctgccagt caagattcag cttcagacgc tccgggcaat aaataatatt ctgcaaaacc
6001 agatcgttaa cggaagcgta ggagtgttta tcatcagcat gaatgtaaaa gagatcgcca
6061 cgggtaatgc gataagggcg atcgttgagt acatgcaggc cattaccgcg ccagacaatc
6121 accagctcac aaaaatcatg tgtatgttca gcaaagacat cttgcggata acggtcagcc
6181 acagcgactg cctgctggtc gctggcaaaa aaatcatctt tgagaagttt taactgatgc
6241 gccaccgtgg ctacctcggc cagagaacga agttgattat tcgcaatatg gcgtacaaat
6301 acgttgagaa gattcgcgtt attgcagaaa gccatcccgt ccctggcgaa tatcacgcgg
6361 tgaccagtta aactctcggc gaaaaagcgt cgaaaagtgg ttactgtcgc tgaatccaca
6421 gcgataggcg atgtcagtaa cgctggcctc gctgtggcgt agcagatgtc gggctttcat
6481 cagtcgcagg cggttcaggt atcgctgagg cgtcagtccc gtttgctgct taagctgccg
6541 atgtagcgta cgcagtgaaa gagaaaattg atccgccacg gcatcccaat tcacctcatc
6601 ggcaaaatgg tcctccagcc aggccagaag caagttgaga cgtgatgcgc tgttttccag
6661 gttctcctgc aaactgcttt tacgcagcaa gagcagtaat tgcataaaca agatctcgcg
6721 actggcggtc gagggtaaat cattttcccc ttcctgctgt tccatctgtg caaccagctg
6781 tcgcacctgc tgcaatacgc tgtggttaac gcgccagtga gacggatact gcccatccag
6841 ctcttgtggc agcaactgat tcagcccggc gagaaactga aatcgatccg gcgagcgata
6901 cagcacattg gtcagacaca gattatcggt atgttcatac agatgccgat catgatcgcg
6961 tacgaaacag accgtgccac aggtgatggt atagggctgc ccattaaaca catgaatacc
7021 cgtgccatgt tcgacaatca caatttcatg aaaatcatga tgatgttcag gaaaatccgc
7081 ctgcgggagc cggggttcta tcgccacgga cgcgttacca gacggaaaaa aatccacact
7141 atgtaatacg gtcatactgg cctcctgatg tcgtcaacac ggcgaaatag taatcacgag
7201 gtcaggttct taccttaaat tttcgacgga aaaccacgta aaaaacgtcg atttttcaag
7261 atacagcgtg aattttcagg aaatgcggtg agcatcacat caccacaatt cagcaaattg
7321 tgaacatcat cacgttcatc tttccctggt tgccaatg
//
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