质粒名称: | BPK2014-AaCas12b |
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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 |
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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 |
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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 |
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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
BPK2014AaCas12b载体基本信息
BPK2014AaCas12b载体图谱质粒图谱和多克隆位点信息
BPK2014AaCas12b载体简介
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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