| 质粒名称: | BPK2014-AaCas12b |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 121949 |
| 存储实验室: | Wei Li |
|
载体骨架基本信息 |
|
| 载体名称: | BPK2014-ccdB |
| 载体出品公司: | -- |
| 载体抗性: | 氯霉素 |
| 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: | -- |
| 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: | AaCas12b,AaC2c1 |
| 物种来源: | Synthetic |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | 10xHis tag （插入件上的C端子） |
| 克隆方法: | 限制性内切酶 |
| 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: | BPK2014-AaCas12b was a gift from Wei Li (Addgene plasmid # 121949 ; http://n2t.net/addgene:121949 ; RRID:Addgene_121949) |
| 参考文献部分: | Repurposing CRISPR-Cas12b for mammalian genome engineering. Teng F, Cui T, Feng G, Guo L, Xu K, Gao Q, Li T, Li J, Zhou Q, Li W. Cell Discov. 2018 Nov 27;4:63. doi: 10.1038/s41421-018-0069-3. eCollection 2018. 10.1038/s41421-018-0069-3 PubMed 30510770 |
买家导航
BPK2014-AaCas12b载体 BPK2014AaCas12b质粒基本信息
BPK2014-AaCas12b质粒图谱 BPK2014AaCas12b载体图谱和BPK2014-AaCas12b载体序列 BPK2014AaCas12b质粒序列多克隆位点信息
BPK2014-AaCas12b质粒 BPK2014AaCas12b载体简介
BPK2014-AaCas12b质粒序列 BPK2014AaCas12b载体序列
LOCUS BPK2014-AaCas12b 7180 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION E. coli expression, protein purification.
ACCESSION .
VERSION .
KEYWORDS BPK2014-AaCas12b
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7180)
AUTHORS Teng F, Cui T, Feng G, Guo L, Xu K, Gao Q, Li T, Li J, Zhou Q, Li W
TITLE Repurposing CRISPR-Cas12b for mammalian genome engineering.
JOURNAL Cell Discov. 2018 Nov 27;4:63. doi: 10.1038/s41421-018-0069-3.
eCollection 2018.
PUBMED 30510770
REFERENCE 2 (bases 1 to 7180)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7180
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 12..36
/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 83..103
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 3503..3532
/codon_start=1
/product="10xHis affinity tag"
/label=10xHis
/translation="HHHHHHHHHH"
promoter 3601..3619
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
terminator 3641..3688
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(3909..4568)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind 4483..4502
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(4569..4671)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
primer_bind complement(5063..5080)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5197..5742)
/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(5277..5296)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5967..7049)
/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 7011..7030
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(7050..7127)
/gene="lacI"
/label=lacI promoter
promoter 7173..11
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 ctcactatag gggaattgtg agcggataac aattcccctg tagaaataat tttgtttaac
61 tttaataagg agatatcata tgcccaagaa gaagaggaag gttcccggga tggccgtgaa
121 gagcatgaag gtgaagctgc gcctggacaa catgcccgag atccgcgccg gcctgtggaa
181 gctgcacacc gaggtgaacg ccggcgtgcg ctactacacc gagtggctga gcctgctgcg
241 ccaggagaac ctgtaccgcc gcagccccaa cggcgacggc gagcaggagt gctacaagac
301 cgccgaggag tgcaaggccg agctgctgga gcgcctgcgc gcccgccagg tggagaacgg
361 ccactgcggc cccgccggca gcgacgacga gctgctgcag ctggcccgcc agctgtacga
421 gctgctggtg ccccaggcca tcggcgccaa gggcgacgcc cagcagatcg cccgcaagtt
481 cctgagcccc ctggccgaca aggacgccgt gggcggcctg ggcatcgcca aggccggcaa
541 caagccccgc tgggtgcgca tgcgcgaggc cggcgagccc ggctgggagg aggagaaggc
601 caaggccgag gcccgcaaga gcaccgaccg caccgccgac gtgctgcgcg ccctggccga
661 cttcggcctg aagcccctga tgcgcgtgta caccgacagc gacatgagca gcgtgcagtg
721 gaagcccctg cgcaagggcc aggccgtgcg cacctgggac cgcgacatgt tccagcaggc
781 catcgagcgc atgatgagct gggagagctg gaaccagcgc gtgggcgagg cctacgccaa
841 gctggtggag cagaagagcc gcttcgagca gaagaacttc gtgggccagg agcacctggt
901 gcagctggtg aaccagctgc agcaggacat gaaggaggcc agccacggcc tggagagcaa
961 ggagcagacc gcccactacc tgaccggccg cgccctgcgc ggcagcgaca aggtgttcga
1021 gaagtgggag aagctggacc ccgacgcccc cttcgacctg tacgacaccg agatcaagaa
1081 cgtgcagcgc cgcaacaccc gccgcttcgg cagccacgac ctgttcgcca agctggccga
1141 gcccaagtac caggccctgt ggcgcgagga cgccagcttc ctgacccgct acgccgtgta
1201 caacagcatc gtgcgcaagc tgaaccacgc caagatgttc gccaccttca ccctgcccga
1261 cgccaccgcc caccccatct ggacccgctt cgacaagctg ggcggcaacc tgcaccagta
1321 caccttcctg ttcaacgagt tcggcgaggg ccgccacgcc atccgcttcc agaagctgct
1381 gaccgtggag gacggcgtgg ccaaggaggt ggacgacgtg accgtgccca tcagcatgag
1441 cgcccagctg gacgacctgc tgccccgcga cccccacgag ctggtggccc tgtacttcca
1501 ggactacggc gccgagcagc acctggccgg cgagttcggc ggcgccaaga tccagtaccg
1561 ccgcgaccag ctgaaccacc tgcacgcccg ccgcggcgcc cgcgacgtgt acctgaacct
1621 gagcgtgcgc gtgcagagcc agagcgaggc ccgcggcgag cgccgccccc cctacgccgc
1681 cgtgttccgc ctggtgggcg acaaccaccg cgccttcgtg cacttcgaca agctgagcga
1741 ctacctggcc gagcaccccg acgacggcaa gctgggcagc gagggcctgc tgagcggcct
1801 gcgcgtgatg agcgtggacc tgggcctgcg caccagcgcc agcatcagcg tgttccgcgt
1861 ggcccgcaag gacgagctga agcccaacag cgagggccgc gtgcccttct gcttccccat
1921 cgagggcaac gagaacctgg tggccgtgca cgagcgcagc cagctgctga agctgcccgg
1981 cgagaccgag agcaaggacc tgcgcgccat ccgcgaggag cgccagcgca ccctgcgcca
2041 gctgcgcacc cagctggcct acctgcgcct gctggtgcgc tgcggcagcg aggacgtggg
2101 ccgccgcgag cgcagctggg ccaagctgat cgagcagccc atggacgcca accagatgac
2161 ccccgactgg cgcgaggcct tcgaggacga gctgcagaag ctgaagagcc tgtacggcat
2221 ctgcggcgac cgcgagtgga ccgaggccgt gtacgagagc gtgcgccgcg tgtggcgcca
2281 catgggcaag caggtgcgcg actggcgcaa ggacgtgcgc agcggcgagc gccccaagat
2341 ccgcggctac cagaaggacg tggtgggcgg caacagcatc gagcagatcg agtacctgga
2401 gcgccagtac aagttcctga agagctggag cttcttcggc aaggtgagcg gccaggtgat
2461 ccgcgccgag aagggcagcc gcttcgccat caccctgcgc gagcacatcg accacgccaa
2521 ggaggaccgc ctgaagaagc tggccgaccg catcatcatg gaggccctgg gctacgtgta
2581 cgccctggac gacgagcgcg gcaagggcaa gtgggtggcc aagtaccccc cctgccagct
2641 gatcctgctg gaggagctga gcgagtacca gttcaacaac gaccgccccc ccagcgagaa
2701 caaccagctg atgcagtgga gccaccgcgg cgtgttccag gagctgctga accaggccca
2761 ggtgcacgac ctgctggtgg gcaccatgta cgccgccttc agcagccgct tcgacgcccg
2821 caccggcgcc cccggcatcc gctgccgccg cgtgcccgcc cgctgcgccc gcgagcagaa
2881 ccccgagccc ttcccctggt ggctgaacaa gttcgtggcc gagcacaagc tggacggctg
2941 ccccctgcgc gccgacgacc tgatccccac cggcgagggc gagttcttcg tgagcccctt
3001 cagcgccgag gagggcgact tccaccagat ccacgccgac ctgaacgccg cccagaacct
3061 gcagcgccgc ctgtggagcg acttcgacat cagccagatc cgcctgcgct gcgactgggg
3121 cgaggtggac ggcgagcccg tgctgatccc ccgcaccacc ggcaagcgca ccgccgacag
3181 ctacggcaac aaggtgttct acaccaagac cggcgtgacc tactacgagc gcgagcgcgg
3241 caagaagcgc cgcaaggtgt tcgcccagga ggagctgagc gaggaggagg ccgagctgct
3301 ggtggaggcc gacgaggccc gcgagaagag cgtggtgctg atgcgcgacc ccagcggcat
3361 catcaaccgc ggcgactgga cccgccagaa ggagttctgg agcatggtga accagcgcat
3421 cgagggctac ctggtgaagc agatccgcag ccgcgtgcgc ctgcaggaga gcgcctgcga
3481 gaacaccggc gacatcgcta gtcatcacca tcaccaccat catcaccatc actaggcggc
3541 cgcataatgc ttaagtcgaa cagaaagtaa tcgtattgta cacggccgca taatcgaaat
3601 taatacgact cactataggg aattcggtac ctgagaataa ctagcataac cccttggggc
3661 ctctaaacgg gtcttgaggg gttttttgct gaaacctcag gcatttgaga agcacacggt
3721 cacactgctt ccggtagtca ataaaccggt aaaccagcaa tagacataag cggctattta
3781 acgaccctgc cctgaaccga cgaccgggtc gaatttgctt tcgaatttct gccattcatc
3841 cgcttattat cacttattca ggcgtagcaa ccaggcgttt aagggcacca ataactgcct
3901 taaaaaaatt acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt
3961 ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc
4021 accttgtcgc cttgcgtata atatttgccc atagtgaaaa cgggggcgaa gaagttgtcc
4081 atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa
4141 aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca
4201 tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat
4261 gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc
4321 accagctcac cgtctttcat tgccatacgg aactccggat gagcattcat caggcgggca
4381 agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag
4441 gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc
4501 tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt
4561 ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt
4621 agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgccgatc aacgtctcat
4681 tttcgccaaa agttggccca gggcttcccg gtatcaacag ggacaccagg atttatttat
4741 tctgcgaagt gatcttccgt cacaggtatt tattcggcgc aaagtgcgtc gggtgatgct
4801 gccaacttac tgatttagtg tatgatggtg tttttgaggt gctccagtgg cttctgtttc
4861 tatcagctgt ccctcctgtt cagctactga cggggtggtg cgtaacggca aaagcaccgc
4921 cggacatcag cgctagcgga gtgtatactg gcttactatg ttggcactga tgagggtgtc
4981 agtgaagtgc ttcatgtggc aggagaaaaa aggctgcacc ggtgcgtcag cagaatatgt
5041 gatacaggat atattccgct tcctcgctca ctgactcgct acgctcggtc gttcgactgc
5101 ggcgagcgga aatggcttac gaacggggcg gagatttcct ggaagatgcc aggaagatac
5161 ttaacaggga agtgagaggg ccgcggcaaa gccgtttttc cataggctcc gcccccctga
5221 caagcatcac gaaatctgac gctcaaatca gtggtggcga aacccgacag gactataaag
5281 ataccaggcg tttccccctg gcggctccct cgtgcgctct cctgttcctg cctttcggtt
5341 taccggtgtc attccgctgt tatggccgcg tttgtctcat tccacgcctg acactcagtt
5401 ccgggtaggc agttcgctcc aagctggact gtatgcacga accccccgtt cagtccgacc
5461 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggaaagacat gcaaaagcac
5521 cactggcagc agccactggt aattgattta gaggagttag tcttgaagtc atgcgccggt
5581 taaggctaaa ctgaaaggac aagttttggt gactgcgctc ctccaagcca gttacctcgg
5641 ttcaaagagt tggtagctca gagaaccttc gaaaaaccgc cctgcaaggc ggttttttcg
5701 ttttcagagc aagagattac gcgcagacca aaacgatctc aagaagatca tcttattaat
5761 cagataaaat atttctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc
5821 agccccatac gatataagtt gtaattctca tgttagtcat gccccgcgcc caccggaagg
5881 agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga
5941 gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
6001 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc
6061 agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg
6121 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt
6181 ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact
6241 accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc
6301 gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc
6361 atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga
6421 atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa
6481 cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg
6541 cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag
6601 acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg
6661 tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc
6721 gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc
6781 agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga
6841 ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg
6901 ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa
6961 acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct
7021 gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg
7081 cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg
7141 ctctccctta tgcgactcct gcattaggaa attaatacga
//
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