| 质粒名称: | 6xHis-MBP-BrCas12b-FNLDTA |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 195340 |
| 存储实验室: | Vladislav Verkhusha |
|
载体骨架基本信息 |
|
| 载体名称: | pET |
| 载体出品公司: | Scott Gradia Lab (Addgene plasmid # 29656) |
| 空载体大小: | 6410bp |
| 完整质粒大小: | 9670bp |
| 载体类型: | Bacterial Expression, CRISPR, Synthetic Biology |
| 载体抗性: | Kanamycin |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | BrCas12b |
| 物种来源: | Brevibacillus sp. SYP-B805 |
| 插入大小: | 3269bp |
| 基因突变: | F208W, N524V, L795I, D868V, T874S, A1015E |
| GenBank编号 | WP_165214399.1 |
| 启动子: | T7 Promoter |
| 融合： |
MBP, TEV cleavage site, 6x His Tag (N terminal on backbone)
6xHis Tag (C terminal on insert)
|
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | TAATACGACTCACTATAGGG |
| 3’测序引物 | GCTAGTTATTGCTCAGCGG |
|
文献引用方法 |
|
| 材料和方法部分: | 6xHis-MBP-BrCas12b-FNLDTA was a gift from Piyush Jain (Addgene plasmid # 195340 ; http://n2t.net/addgene:195340 ; RRID:Addgene_195340) |
| 参考文献部分: | Engineering highly thermostable Cas12b via de novo structural analyses for one-pot detection of nucleic acids. Nguyen LT, Rananaware SR, Yang LG, Macaluso NC, Ocana-Ortiz JE, Meister KS, Pizzano BLM, Sandoval LSW, Hautamaki RC, Fang ZR, Joseph SM, Shoemaker GM, Carman DR, Chang L, Rakestraw NR, Zachary JF, Guerra S, Perez A, Jain PK. Cell Rep Med. 2023 May 16;4(5):101037. doi: 10.1016/j.xcrm.2023.101037. Epub 2023 May 8. 10.1016/j.xcrm.2023.101037 PubMed 37160120 |
买家导航
6xHis-MBP-BrCas12b-FNLDTA载体质粒基本信息
6xHis-MBP-BrCas12b-FNLDTA质粒图谱载体图谱和6xHis-MBP-BrCas12b-FNLDTA载体序列质粒序列多克隆位点信息
6xHis-MBP-BrCas12b-FNLDTA质粒载体简介
6xHis-MBP-BrCas12b-FNLDTA质粒序列载体序列
LOCUS 6xHis-MBP-BrCas12b-FNLDTA 9670 bp ds-DNA circular SYN 27-FEB-2023
DEFINITION Expresses Engineered BrCas12b in Bacteria.
ACCESSION .
VERSION .
KEYWORDS 6xHis-MBP-BrCas12b-FNLDTA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9670)
TITLE Engineering Highly Thermostable Cas12b via De Novo Structural
Analyses for One-Pot Detection of Nucleic Acids
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 9670)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Feb 27, 2023 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..9670
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 106..125
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 209..228
/label=T7
/note="T7 promoter, forward primer"
promoter 209..227
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 228..252
/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 267..289
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 308..325
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 335..1435
/codon_start=1
/gene="malE (mutated)"
/product="maltose binding protein from E. coli"
/label=MBP
/note="This version of the gene does not encode a signal
sequence, so MBP will remain in the cytosol."
/translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
WYAVRTAVINAASGRQTVDEALKDAQT"
primer_bind 1409..1432
/label=MBP-F
/note="Maltose binding protein, forward primer"
CDS 1448..1468
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQS"
CDS 4742..4759
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(4812..4830)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 4826..4873
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 4910..5365
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4997..5016)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5207..5228
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(5458..6273)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 6181..6200
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 6265..6284
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 6395..6983
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6884..6903
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7137..7154
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 7169..7308
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(7234..7253)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 7394..7416
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(7410..7601)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(8410..9492)
/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 9454..9473
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(9493..9570)
/gene="lacI"
/label=lacI promoter
ORIGIN
1 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
61 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
121 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
181 gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg
241 gataacaatt cccctctaga aataattttg tttaacttta agaaggagat ataccatggg
301 ttcttctcac catcaccatc accatggttc ttctatgaaa atcgaagaag gtaaactggt
361 aatctggatt aacggcgata aaggctataa cggtctcgct gaagtcggta agaaattcga
421 gaaagatacc ggaattaaag tcaccgttga gcatccggat aaactggaag agaaattccc
481 acaggttgcg gcaactggcg atggccctga cattatcttc tgggcacacg accgctttgg
541 tggctacgct caatctggcc tgttggctga aatcaccccg gacaaagcgt tccaggacaa
601 gctgtatccg tttacctggg atgccgtacg ttacaacggc aagctgattg cttacccgat
661 cgctgttgaa gcgttatcgc tgatttataa caaagatctg ctgccgaacc cgccaaaaac
721 ctgggaagag atcccggcgc tggataaaga actgaaagcg aaaggtaaga gcgcgctgat
781 gttcaacctg caagaaccgt acttcacctg gccgctgatt gctgctgacg ggggttatgc
841 gttcaagtat gaaaacggca agtacgacat taaagacgtg ggcgtggata acgctggcgc
901 gaaagcgggt ctgaccttcc tggttgacct gattaaaaac aaacacatga atgcagacac
961 cgattactcc atcgcagaag ctgcctttaa taaaggcgaa acagcgatga ccatcaacgg
1021 cccgtgggca tggtccaaca tcgacaccag caaagtgaat tatggtgtaa cggtactgcc
1081 gaccttcaag ggtcaaccat ccaaaccgtt cgttggcgtg ctgagcgcag gtattaacgc
1141 cgccagtccg aacaaagagc tggcaaaaga gttcctcgaa aactatctgc tgactgatga
1201 aggtctggaa gcggttaata aagacaaacc gctgggtgcc gtagcgctga agtcttacga
1261 ggaagagttg gcgaaagatc cacgtattgc cgccactatg gaaaacgccc agaaaggtga
1321 aatcatgccg aacatcccgc agatgtccgc tttctggtat gccgtgcgta ctgcggtgat
1381 caacgccgcc agcggtcgtc agactgtcga tgaagccctg aaagacgcgc agactaatgg
1441 gatcgaggaa aacctgtact tccaatccaa tatgcccgta aggtcattta aagttaagct
1501 agtgactcgg tctggcgatg cggagcacat gctgcaactg cgtagggggc tgtggaaaac
1561 ccatgaaatc gtaaatcagg gcattgcgta ctacatgaac aagttggcgc tgatgcgtca
1621 agaaccgtat gctggtaaaa gccgtgaggt tgttcgtttg gagctgctgc atagcctgcg
1681 tgcacagcag aaacgtaata actggaccgg cgacgccggt acggatgatg aaattctgaa
1741 cctgagccgt cgcctgtacg agttgctcgt gccgagcgcg atcggcgaga agggcgatgc
1801 gcagatgtta tctcgcaagt ttctgtcgcc gctggttgat ccgaattctg aaggtggtaa
1861 aggcacggcg aagtccggcc gtaagccgcg ttggatgaaa atgcgggagg agggtcatcc
1921 ggattgggaa gctgaacgcg agaaggaccg cgcaaagaaa gcagcggacc cgactgcgtc
1981 tattctgaat gatttggaag cgtttggtct gcgtccgctg ttcccgctgt tcaccgacga
2041 gcaaaaaggc atccagtggc tgccgaaaca aaaacgtcag ttcgtgcgta cctgggatcg
2101 tgatatgttt cagcaggctc tggagcgtat gctgtcctgg gagtcctgga accgccgtgt
2161 cgccgaggaa tatcagaagc tgcaggctca acgtgacgag ttgtacgcaa aatatctggc
2221 cgatggtggt gcctggctgg aagcgttaca aagctttgag aagcagcgtg aagttgaact
2281 ggctgaggag tcttttgccg ctaaatccga atatctgatc acccgacgac aaattcgcgg
2341 ttggaaacag gtgtacgaaa agtggtcaca actgccagag cacgccgccc aagaacaatt
2401 ctggcaagtt gtagcggacg tacagacgtc tctgcctggt gcgtttggtg acccaaaagt
2461 ttaccagttt ctgagccagc cggagcatca ccacatctgg cgcggttacc cgaaccgact
2521 attccactat tctgattaca acggtgttcg caagaagctg caaagagcgc gtcacgatgc
2581 gacttttacc ttgccggacc cggttgaaca cccgctttgg atcagattcg atgcgcgtgg
2641 tggtaatatt catgattatg agatcagcca gaatggcaag caatatcaag ttaccttctc
2701 ccgcctcctg tggcctgaaa atgaaacctg ggtggagcgt gaaaacgtta ccgttgcaat
2761 cggggcaagc cagcagctta agcgtcagat tcggttggac ggctacgccg ataaaaagca
2821 gaaggtacgt tatcgtgact actctagcgg catcgagctg accggtgttc tgggtggtgc
2881 gaagatccag ttcgaccgtc gtcacctgcg caaagcaagt aatcgtttag ccgacggtga
2941 aaccggccct gtgtacctga acgtggtggt cgatatcgag ccgtttctgg ctatgcgcaa
3001 cggtcgtcta caaacgccga ttggccaagt gctgcaagtc gtgaccaagg actggccgaa
3061 ggtgacgggt tataaaccgg ctgagctgat cagctggatt caaaacagcc cgctggcggt
3121 gggtacaggc gtcaacacca ttgaggcggg tatgcgcgtt atgagtgttg atctgggtca
3181 aagaagcgca gctgctgtga gcattttcga ggtcatgcgc cagaaaccgg cggaacagga
3241 aaccaaactc ttctatccga ttgctgtgac cggtctgtat gcggtgcatc gtcgtagttt
3301 gctgctgcgt cttccaggcg agaaaattag cgacgaaatc gaacagcaac gtaaaatcag
3361 agcccatgct cgtagcttgg tgcgctacca gatccgtctg cttgcggacg tactgcgtct
3421 ccacacccgt ggcaccgcgg aacagagacg tgcgaaactg gatgaattgc tggctaccct
3481 gcaaacgaaa caagagttag atcagaagtt atggcaaact gagttggaaa agctgttcga
3541 ctacattcac gaaccggcgg agcgctggca gcaggcgctg gtggcggcgc accgtacctt
3601 ggagccggtt atcggtcagg cagttcgtca ttggcgtaag tccctccgca ttgatcgtaa
3661 gggcttggcg ggcatgagca tgtggaacat tgaagagttg gaggaaaccc ggaagttgtt
3721 gatcgcatgg agcaaacatt ccagggtccc cggcgagccg aaccgtctgg acaaagaaga
3781 aacgtttgca ccgcagcagc tccaacatat ccaaaacgtc aaagacgacc gcttgaagca
3841 gatggcaaat ctgattgtga tgaccgcctt gggctacaag tacgatgagg ccgagaaaca
3901 atggaaagag gcgtatccgg cgtgtcagat gattctgttc gaagatctaa gccgctaccg
3961 ctttgcgctg gaccgaccgc gtcgcgagaa caaccggtta atgaaatggg cacacagaag
4021 tatcccgcgt ctggtttatc tgcaaggtga attgttcggc atccaggtgg gtgtggttta
4081 ttcggcgtat agcagccgtt tccatgctaa aactggtgcg ccaggcattc gctgccacgc
4141 gctgaaagaa gaagacctgc aaccgaacag ctacgtggtg aagcaactga ttaaagacgg
4201 ctttatccgt gaggaccaga ccggttccct taagccgggt cagattgtac cgtggtctgg
4261 cggtgaactg ttcgttacct tagccgaccg ttccggcagc cgtttggccg tgatccacgc
4321 cgacatcaat gcagcgcaga atctgcaaaa gagattctgg caacagaaca ccgagatctt
4381 tcgtgtgccg tgcaaagtga ctacgtccgg tctgataccg gcatacgaca agatgaaaaa
4441 gttattcggc aaaggctatt tcgcaaagat caaccagacg gacaccagcg aagtgtacgt
4501 ttgggaacac agcgaaaaga tgaagggcaa aacgaccccg gcggacccag cagaagaagg
4561 agtttttgat gagtccctga ccgacgagat ggaagagctt gaagacagcc aagagggtta
4621 caagaccctg ttccgtgatc cgtcgggctt tttttggtcg agcgatcgct ggctgccgca
4681 gaaagagttc tggttttggg ttaaacgccg tatcgagaag aaactgcgcg agcagctcca
4741 gcaccaccac caccaccact gagatccggc tgctaacaaa gcccgaaagg aagctgagtt
4801 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
4861 gaggggtttt ttgctgaaag gaggaactat atccggattg gcgaatggga cgcgccctgt
4921 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
4981 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
5041 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
5101 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
5161 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
5221 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
5281 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
5341 aacaaactag taacgtttac aatttcaggt ggcacttttc ggggaaatgt gcgcggaacc
5401 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgaa ttaattctta
5461 gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc
5521 atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag
5581 gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat
5641 taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga
5701 atccggtgag aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc
5761 attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc
5821 ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg
5881 caaccggcgc aggaacactg ccagcgcatc aacaatgttt tcacctgaat caggatattc
5941 ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc
6001 aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag
6061 tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa
6121 ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt
6181 atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta atcgcggcct
6241 agagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta
6301 agcagacagt tttattgttc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
6361 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
6421 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
6481 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
6541 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
6601 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
6661 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
6721 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
6781 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
6841 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
6901 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
6961 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
7021 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
7081 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
7141 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
7201 gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
7261 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
7321 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
7381 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
7441 gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc
7501 tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata
7561 aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg
7621 ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg
7681 gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta
7741 tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca
7801 gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg
7861 gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt
7921 catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt
7981 atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac
8041 gacaggagca cgatcatgcg cacccgtggg gccgccatgc cggcgataat ggcctgcttc
8101 tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt
8161 ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg
8221 aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc
8281 ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag
8341 gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt
8401 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
8461 atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt
8521 caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag
8581 caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg
8641 cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc
8701 aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc
8761 aaccagcatc gcagtgggaa cgataccctc attcagcatt tgcatggttt gttgaaaacc
8821 ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag
8881 atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa
8941 cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc
9001 ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc
9061 cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt
9121 aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc
9181 gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga
9241 tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc
9301 aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag
9361 ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt
9421 caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt
9481 tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc
9541 gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact
9601 cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga
9661 atggtgcatg
//
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6xHis-MBP-BrCas12b-FNLDTA质粒载体应用举例
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