| 质粒名称: | pC0075 TtCsm6 His6-TwinStrep-SUMO-BsaI |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 115270 |
| 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: | 氨苄青霉素 |
| 空载体大小: | 6084 bp |
| 完整质粒大小: | 7479 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: | TtCsm6,Csm6 from Thermus thermophilus |
| 物种来源: | Thermus thermophilus |
| 插入大小: | 1395 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: | pC0075 TtCsm6 His6-TwinStrep-SUMO-BsaI was a gift from Feng Zhang (Addgene plasmid # 115270 ; http://n2t.net/addgene:115270 ; RRID:Addgene_115270) |
| 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 pC0075 TtCsm6 His6-TwinStrep-SUMO-BsaI 7476 bp ds-DNA circular SYN 20-SEP-2018
DEFINITION Expresses TtCsm6 from a SUMO-tagged bacterial expression construct.
ACCESSION .
VERSION .
KEYWORDS pC0075 TtCsm6 His6-TwinStrep-SUMO-BsaI
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7476)
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 7476)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 20, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7476
/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(1958..1976)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 1972..2019
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter complement(2194..2222)
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
primer_bind complement(2249..2267)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 2335..2439
/gene="bla"
/label=AmpR promoter
CDS 2440..3300
/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(2658..2677)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 3471..4059
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3960..3979
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4213..4230
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 4245..4384
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(4310..4329)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4470..4492
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(4486..4677)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(5989..7071)
/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 7033..7052
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(7072..7149)
/gene="lacI"
/label=lacI promoter
primer_bind 7355..7374
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 7458..1
/label=T7
/note="T7 promoter, forward primer"
promoter 7458..7476
/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 ggaggacctt gatgcgttgt
541 gggagcgcta tcgtgaagct gttcgcgcgg gtgggaatcc ccaggctctg taccaggaga
601 tggtatggcc ggcgttgtta gcactgtggc gcgagaaacc tcgcgtttac cccttccctc
661 aagcatttgc ggtgtcagtg catacccttg gaacgagccc ggaggcaact gctcttgcga
721 tcttaggtgc aggcgccgag cgcgtttatg tgttacacac tccggagtca gcgcgcttct
781 tgccgcgctt acgtcaagac accggaaagg acttgtaccc agttgagatt ggaaagagtg
841 acgtagaggc gatctatcgt gaagtgaagc gcttattgga aaagcacccc gaagtccctg
901 tcgcattgga cctgacttcg gggaccaagg caatgtcggc agggttggcc gccgcaggat
961 tttttttcca acgtttttat cccaaagtcc gtgtggtgta tgtagataac gaagactatg
1021 acccagaatt gcgccgcccg cgcgccggga cggagaaact tcgcattttg cccaatccac
1081 acgaggcttt ggctgaagtc gacgcattgt ttgccaaaga gctttatggc aaaggagaat
1141 ttgggcaggc ggcggcttac tttcgcggaa tggttggtcg cactggtaat caagcatacg
1201 cattatatgc attgctggcg gagatgtacc gcgcctggcg cgccttggat ttcggtgaag
1261 cattaaaggc ggggcgtaag ctgctgggac aactgtctca aaatgtgtgg ttgaatcatc
1321 cactgaacgc ccgtcgcgaa gcgcttgagg cgcaggtcgc attattagag gccgtcgacc
1381 gctttttgaa ggcgcgtgac ttcgcattaa aggaaggcgt gtatggtctg gcccgcactc
1441 ttcttcatct ggcccaagag gcgaaagaag aagccgcggt acttgcagca ttgtatgcat
1501 accgcgcatt ggagcttctt ttacaggagc gcttagctct gcttggtcgt cgcgcggagg
1561 cgcctggtct gagcccagaa gaagccgagg cattgcgcaa ggctttagcc gagctgttag
1621 gagttcttcc tgaagaggtt cgtcttccag ctaaattagg tttattagat ttattggcct
1681 ttttgcgctt gaaaggagat gaggcgttgg gccgcctgtc tctggctgaa ctgcgtggct
1741 tagccggtgc tcttaaaggc cgtaattcag cccttcttgt acatgggttt gatgtgccca
1801 gtcctaaggc cgtcgagggc atcgcccgtc tggcgcaagg tcttctgcag gatcttgagg
1861 cacgtacggc tctgggcccg ctgtctccgg aacccgtacc cttggggttc taaggccgca
1921 ctcgaggccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
1981 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
2041 ggatatcccg caagaggccc ggcagtaccg gcataaccaa gcctatgcct acagcatcca
2101 gggtgacggt gccgaggatg acgatgagcg cattgttaga tttcatacac ggtgcctgac
2161 tgcgttagca atttaactgt gataaactac cgcattaaag cttatcgatg ataagctgtc
2221 aaacatgaga attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
2281 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
2341 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
2401 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
2461 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
2521 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
2581 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
2641 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
2701 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
2761 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
2821 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
2881 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
2941 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg
3001 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
3061 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
3121 tttattgctg ataaatctgg agccggtgag cgtgggtcac gcggtatcat tgcagcactg
3181 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
3241 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
3301 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
3361 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
3421 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
3481 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
3541 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
3601 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
3661 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
3721 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
3781 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
3841 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
3901 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
3961 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
4021 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
4081 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
4141 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
4201 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
4261 ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct
4321 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
4381 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
4441 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
4501 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga
4561 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa
4621 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga
4681 tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag
4741 gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg
4801 taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca
4861 gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca
4921 gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg
4981 gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct
5041 tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc
5101 tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct
5161 gcccgagatg cgccgcgtgc ggctgctgga gatggcggac gcgatggata tgttctgcca
5221 agggttggtt tgcgcattca cagttctccg caagaattga ttggctccaa ttcttggagt
5281 ggtgaatccg ttagcgaggt gccgccggct tccattcagg tcgaggtggc ccggctccat
5341 gcaccgcgac gcaacgcggg gaggcagaca aggtataggg cggcgcctac aatccatgcc
5401 aacccgttcc atgtgctcgc cgaggcggca taaatcgccg tgacgatcag cggtccaatg
5461 atcgaagtta ggctggtaag agccgcgagc gatccttgaa gctgtccctg atggtcgtca
5521 tctacctgcc tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc ggaagcgaga
5581 agaatcataa tggggaaggc catccagcct cgcgtcgcga acgccagcaa gacgtagccc
5641 agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg
5701 ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg
5761 ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc
5821 ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc
5881 atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga
5941 gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
6001 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
6061 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
6121 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
6181 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
6241 ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt
6301 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
6361 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
6421 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
6481 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
6541 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
6601 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
6661 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
6721 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
6781 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
6841 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
6901 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
6961 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
7021 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
7081 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
7141 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
7201 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
7261 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
7321 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
7381 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
7441 tctcgatccc gcgaaattaa tacgactcac tatagg
//
