| 质粒名称: | pC0073 EiCsm6 His6-TwinStrep-SUMO-BsaI |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 115268 |
| 存储实验室: | Feng Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | SUMO bacterial expression vector |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄霉素 |
| 空载体大小: | 6084 bp |
| 完整质粒大小: | 7377 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | EiCsm6 |
| 物种来源: | 意大利肠球菌 |
| 插入大小: | 1293 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | SUMO |
| 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: | pC0073 EiCsm6 His6-TwinStrep-SUMO-BsaI was a gift from Feng Zhang (Addgene plasmid # 115268 ; http://n2t.net/addgene:115268 ; RRID:Addgene_115268) |
| 参考文献部分: | Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6. Gootenberg JS, Abudayyeh OO, Kellner MJ, Joung J, Collins JJ, Zhang F. Science. 2018 Apr 27;360(6387):439-444. doi: 10.1126/science.aaq0179. Epub 2018 Feb 15. 10.1126/science.aaq0179 PubMed 29449508 |
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 pC0073 EiCsm6 His6-TwinStrep-SUMO-BsaI 7374 bp ds-DNA circular SYN 21-SEP-2018
DEFINITION Expresses EiCsm6 from a SUMO-tagged bacterial expression construct.
ACCESSION .
VERSION .
KEYWORDS pC0073 EiCsm6 His6-TwinStrep-SUMO-BsaI
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7374)
AUTHORS Gootenberg JS, Abudayyeh OO, Kellner MJ, Joung J, Collins JJ, Zhang
F
TITLE Multiplexed and portable nucleic acid detection platform with Cas13,
Cas12a, and Csm6.
JOURNAL Science. 2018 Apr 27;360(6387):439-444. doi:
10.1126/science.aaq0179. Epub 2018 Feb 15.
PUBMED 29449508
REFERENCE 2 (bases 1 to 7374)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 21, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7374
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 1..25
/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 40..62
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 81..98
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 108..125
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
CDS 138..161
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
CDS 198..221
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
CDS 222..512
/codon_start=1
/gene="S. cerevisiae SMT3 (truncated)"
/product="cleavable ubiquitin-like protein tag"
/label=SUMO
/translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIG"
primer_bind complement(1856..1874)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 1870..1917
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter complement(2092..2120)
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
primer_bind complement(2147..2165)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 2233..2337
/gene="bla"
/label=AmpR promoter
CDS 2338..3198
/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(2556..2575)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 3369..3957
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3858..3877
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4111..4128
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 4143..4282
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(4208..4227)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4368..4390
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(4384..4575)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(5887..6969)
/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 6931..6950
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(6970..7047)
/gene="lacI"
/label=lacI promoter
primer_bind 7253..7272
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 7356..1
/label=T7
/note="T7 promoter, forward primer"
promoter 7356..7374
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga
61 gatataccat gggcagcagc catcatcatc atcatcacag cagcggcctg gtgccgcgcg
121 gcagccatat ggctagctgg agccatccgc agtttgaaaa aggtggtggt agcggtggtg
181 gttcaggtgg tagtgcatgg tcacaccctc agtttgagaa aatgtcggac tcagaagtca
241 atcaagaagc taagccagag gtcaagccag aagtcaagcc tgagactcac atcaatttaa
301 aggtgtccga tggatcttca gagatcttct tcaagatcaa aaagaccact cctttaagaa
361 ggctgatgga agcgttcgct aaaagacagg gtaaggaaat ggactcctta agattcttgt
421 acgacggtat tagaatccaa gctgatcaga cccctgaaga tttggacatg gaggataacg
481 atattattga ggctcacaga gaacagattg gtggatctat gaagattctt tttagcccga
541 ttggcaacac agatccttgg cgcaatgacc gcgatggtgc gatgttgcac atcgttcgtc
601 actaccaacc tgaccgtgta gtgctttttt ttaccgagtc catctggcaa gggaatcaac
661 acttctcagg acaacaagcc tttgattggg tgaaaatcat tcagagtatt aacgagaact
721 gtcagattga aattaaatgt gatactattg aggtggagaa tgatttcgat gcctacaaag
781 atttgttcca tcagtactta gtcgaggaaa aacgtaaata tcctaatgct gaaattttcc
841 tgaatgtcac aagcgggact ccgcagatgg aaacaacttt gtgccttgaa tatgtgactt
901 accccgacaa aatgcgctgc attcaggtca gcaccccgct taagactagc aatgctaaaa
961 ccaaatacgc gcaagccgat tgccaggaag ttgaccttga aatcgttaat gaagaggagt
1021 ctcagcaacc ttcacgctgt cacaagatcg ctattctgag ctttcgtgaa gcaatcgtac
1081 gtaatcagat caaatcatta cttgataatt acgactacga agccgcgctg cagcttgttg
1141 catcacaaaa gagtttccgc aatgggaagg agattcgcaa aaagctgaaa gagctgatcg
1201 acgacattaa gatgcaccgt gtgttctcat acttaatcaa acagtatccc cgcaacgaga
1261 agttgcaaaa agctctgtta cacaccattc ttttagaaat gcgtcaccaa cgtggagaca
1321 ttgccgaaac tctgatccgt gtaaagagta ttgcagagta tatcgtggaa caatatatcc
1381 aaaagaacta tccatatctg atcatctaca aagaagacaa gccttacttc aacgttagct
1441 acagtcaaga gctgaccgag agttatttgg ccctgatgga cagccgcaac aagaagacta
1501 acaagaaaat gaccgtggac tccctggatc gtatcttggg gtttcccgct taccgcgatt
1561 tcttgcaact gttagaagcc agcaacgaga tgactaacga gatgaataaa gtaaatgaaa
1621 ttaataacct tcgcaataag gtcgcccata acttagacag tcttaattta gatcgtgaca
1681 agaatggtcg caaaatcacg aatgcggtaa ctgcggtgcg caccatgctg ttggcggtct
1741 tcccagaggt tcaggaaaac gacttccatt atttaaagca atttaatcag agcattaaag
1801 aacttttgta aggccgcact cgaggcccga aaggaagctg agttggctgc tgccaccgct
1861 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
1921 aaaggaggaa ctatatccgg atatcccgca agaggcccgg cagtaccggc ataaccaagc
1981 ctatgcctac agcatccagg gtgacggtgc cgaggatgac gatgagcgca ttgttagatt
2041 tcatacacgg tgcctgactg cgttagcaat ttaactgtga taaactaccg cattaaagct
2101 tatcgatgat aagctgtcaa acatgagaat tcttgaagac gaaagggcct cgtgatacgc
2161 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
2221 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
2281 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
2341 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
2401 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
2461 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
2521 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
2581 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
2641 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
2701 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
2761 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
2821 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
2881 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
2941 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
3001 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtcacgc
3061 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
3121 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
3181 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
3241 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
3301 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
3361 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
3421 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
3481 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
3541 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
3601 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
3661 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
3721 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
3781 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
3841 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
3901 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
3961 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
4021 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
4081 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
4141 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca
4201 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
4261 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
4321 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
4381 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
4441 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
4501 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
4561 ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat
4621 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
4681 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat
4741 cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca
4801 gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc
4861 cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt
4921 tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt
4981 aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg
5041 tggccaggac ccaacgctgc ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc
5101 gatggatatg ttctgccaag ggttggtttg cgcattcaca gttctccgca agaattgatt
5161 ggctccaatt cttggagtgg tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc
5221 gaggtggccc ggctccatgc accgcgacgc aacgcgggga ggcagacaag gtatagggcg
5281 gcgcctacaa tccatgccaa cccgttccat gtgctcgccg aggcggcata aatcgccgtg
5341 acgatcagcg gtccaatgat cgaagttagg ctggtaagag ccgcgagcga tccttgaagc
5401 tgtccctgat ggtcgtcatc tacctgcctg gacagcatgg cctgcaacgc gggcatcccg
5461 atgccgccgg aagcgagaag aatcataatg gggaaggcca tccagcctcg cgtcgcgaac
5521 gccagcaaga cgtagcccag cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg
5581 ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg
5641 aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa
5701 atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata
5761 agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
5821 ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg
5881 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
5941 ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac
6001 cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa
6061 gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg
6121 gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac
6181 gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac
6241 cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga
6301 catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata
6361 tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag
6421 cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc
6481 atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg
6541 aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat
6601 gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac
6661 gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt
6721 aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat
6781 cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc
6841 cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac
6901 cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac
6961 tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg
7021 aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct
7081 gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg
7141 gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
7201 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
7261 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
7321 cggcgtagag gatcgagatc tcgatcccgc gaaattaata cgactcacta tagg
//
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