| 质粒名称: | cBEST4 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 234660 |
| 存储实验室: | Mingzi M. Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | pMDY23 |
| 载体出品公司: | Clontech |
| 空载体大小: | 9821bp |
| 完整质粒大小: | 9821bp |
| 载体类型: | Bacterial Expression, CRISPR, Synthetic Biology |
| 载体抗性: | Chloramphenicol, 25 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称1: | spCas9 cytosine base editor、APOBEC1-spCas9n-UGI fusion protein |
| 物种来源: | Synthetic |
| 插入大小: | 5100bp |
| 启动子: | P3 |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | EcoRI (not destroyed) |
| 3’克隆位点: | SpeI (not destroyed) |
| 5’测序引物 | GCAATTAACCCTCACTAAAGG |
| 插入基因名称2: | Golden Gate compatible sgRNA insertion cassette |
| 物种来源: | Synthetic |
| 启动子: | tcp830 |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | SpeI (not destroyed) |
| 3’克隆位点: | NotI (not destroyed) |
| 5’测序引物 | GGGAAACGCCTGGTATCTTT |
|
文献引用方法 |
|
| 材料和方法部分: | cBEST4 was a gift from Mingzi M. Zhang (Addgene plasmid # 234660 ; http://n2t.net/addgene:234660 ; RRID:Addgene_234660) |
| 参考文献部分: | Highly efficient CRISPR-Cas9 base editing in Bifidobacterium with bypass of restriction modification systems. Lin H-C, Hsiao W-C, Hsu Y-C, Lin M-C, Hsu C-C, Zhang MM. Appl Environ Microbiol. 2025 Mar 10:e0198524. doi: 10.1128/aem.01985-24. 10.1128/aem.01985-24 PubMed 40062897 |
买家导航
cBEST4载体质粒基本信息
cBEST4质粒图谱载体图谱和cBEST4载体序列质粒序列多克隆位点信息
cBEST4质粒载体简介
cBEST4质粒序列载体序列
LOCUS cBEST4 9821 bp DNA circular SYN 15-DEC-2025
DEFINITION Expresses cytosine base editor - spCas9n (D10A) fused to APOBEC1 and
UGI, Include Golden Gate compatible cassette for sgRNA insertion.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9821)
AUTHORS Lin H-C, Hsiao W-C, Hsu Y-C, Lin M-C, Hsu C-C, Zhang MM
TITLE Highly efficient CRISPR-Cas9 base editing in Bifidobacterium with
bypass of restriction modification systems.
JOURNAL Appl Environ Microbiol. 2025 Mar 10:e0198524. doi:
10.1128/aem.01985-24.
PUBMED 40062897
REFERENCE 2 (bases 1 to 9821)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Dec 15, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Appl
Environ Microbiol. 2025 Mar 10:e0198524. doi: 10.1128/aem.01985-24."
COMMENT Sequence Label: cBEST4
FEATURES Location/Qualifiers
source 1..9821
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 4..22
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 49..333
/label=gapdh promoter
/note="glyceraldehyde-3-phosphate dehydrogenase promoter
from Eggerthella lenta"
CDS 334..1020
/codon_start=1
/product="cytidine deaminase (C to U editing enzyme) from
rat"
/label=APOBEC-1
/note="can use ssDNA as a substrate (Komor et al., 2016)"
/translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
LWATGLK"
CDS join(1069..1092,1093..1095,1096..5169)
/codon_start=1
/product="nickase mutant of the Cas9 endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system"
/label=Cas9(D10A)
/note="generates RNA-guided single strand nicks in DNA due
to the D10A mutation in the RuvC catalytic domain"
/translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 5182..5433
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
terminator complement(5614..5662)
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
protein_bind 5802..5820
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 5976..5998
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 5990..6007
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 5991..6007
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 6017..6036
/label=T7
/note="T7 promoter, forward primer"
promoter 6017..6035
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(6083..6101)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(6122..6138)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6122..6138)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6135..6157)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6146..6162
/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)."
promoter complement(join(6170..6176,6177..6194,6195..6200))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6215..6236
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
misc_RNA 6278..6353
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
terminator 6375..6409
/label=lambda t0 terminator
/note="minimal transcription terminator from phage lambda
(Scholtissek and Grosse, 1987)"
rep_origin complement(6451..7039)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6531..6550)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tgcaattaac cctcactaaa gggaacaaaa gctggagctc cagaattcgc tgctccttcg
61 gtcggacgtg cgtctacggg caccttaccg cagccgtcgg ctgtgcgaca cggacggatc
121 gggcgaactg gccgatgctg ggagaagcgc gctgctgtac ggcgcgcacc gggtgcggag
181 cccctcggcg agcggtgtga aacttctgtg aatggcctgt tcggttgctt tttttatacg
241 gctgccagat aaggcttgca gcatctgggc ggctaccgct atgatcgggg cgttcctgca
301 attcttagtg cgagtatctg aaaggggata cgcatgtcct ccgagaccgg ccccgtggcc
361 gtggacccga ccctgcgtcg ccgcatcgag ccgcacgagt tcgaggtgtt cttcgacccg
421 cgcgagctgc gcaaggagac ctgcctgctg tacgagatca actggggcgg ccggcactcc
481 atctggcgcc acacctcgca gaacaccaac aagcacgtcg aggtcaactt catcgagaag
541 ttcaccaccg agcggtactt ctgcccgaac acccgctgct ccatcacctg gttcctgtcc
601 tggtccccct gcggcgagtg ctcccgggcc atcaccgagt tcctgtcccg ctacccgcac
661 gtcaccctgt tcatctacat cgcccggctg taccaccacg ccgacccgcg caaccggcag
721 ggcctgcgcg acctgatctc ctccggcgtc accatccaga tcatgaccga gcaggagtcc
781 ggctactgct ggcggaactt cgtcaactac tccccgtcca acgaggccca ctggccccgc
841 tacccgcacc tgtgggtccg cctgtacgtc ctggagctgt actgcatcat cctgggcctg
901 ccgccctgcc tgaacatcct gcgtcgcaag cagccgcagc tgaccttctt caccatcgcc
961 ctccagtcct gccactacca gcgcctgccg ccccacatcc tgtgggccac cggcctgaag
1021 tccggctccg agaccccggg cacctcggag tccgccaccc cggagtccga caagaagtac
1081 tccatcggcc tcgcgatcgg caccaactcc gtgggctggg cggtcatcac cgacgagtac
1141 aaggtcccct ccaagaagtt caaggtcctg ggcaacaccg accggcactc gatcaagaag
1201 aacctgatcg gcgccctgct cttcgacagc ggcgagaccg ccgaggcgac ccgcctgaag
1261 cggaccgcgc gtcgccgcta cacccggcgc aagaaccgca tctgctacct gcaggaaatc
1321 ttctccaacg agatggccaa ggtggacgac tcgttcttcc accgcctgga ggagagcttc
1381 ctggtggagg aggacaagaa gcacgagcgc cacccgatct tcggcaacat cgtggacgag
1441 gtggcctacc acgagaagta ccccaccatc taccacctcc gcaagaagct ggtggactcg
1501 accgacaagg cggacctgcg gctcatctac ctggccctcg cgcacatgat caagttccgc
1561 ggccacttcc tcatcgaggg cgacctgaac ccggacaact ccgacgtgga caagctcttc
1621 atccagctgg tgcagaccta caaccagctg ttcgaggaga accccatcaa cgccagcggc
1681 gtggacgcca aggcgatcct ctccgcgcgc ctgagcaagt cccggcgcct ggagaacctc
1741 atcgcccagc tgccgggcga gaagaagaac ggcctcttcg gcaacctgat cgcgctgtcg
1801 ctcggcctga cccccaactt caagagcaac ttcgacctgg ccgaggacgc gaagctccag
1861 ctgtccaagg acacctacga cgacgacctg gacaacctgc tcgcccagat cggcgaccag
1921 tacgcggacc tcttcctggc cgcgaagaac ctctcggacg ccatcctgct cagcgacatc
1981 ctgcgggtca acaccgagat caccaaggcc ccgctgtcgg cgagcatgat caagcggtac
2041 gacgagcacc accaggacct gaccctgctc aaggccctcg tgcgccagca gctgcccgag
2101 aagtacaagg aaatcttctt cgaccagtcc aagaacggct acgccggcta catcgacggc
2161 ggcgcgtcgc aggaggagtt ctacaagttc atcaagccga tcctggagaa gatggacggc
2221 accgaggagc tgctcgtcaa gctgaaccgc gaggacctgc tccgcaagca gcggaccttc
2281 gacaacggct ccatcccgca ccagatccac ctgggcgagc tccacgccat cctccggcgc
2341 caggaggact tctacccctt cctgaaggac aaccgcgaga agatcgagaa gatcctgacc
2401 ttccgcatcc cgtactacgt cggccccctg gcccgcggca actcccggtt cgcgtggatg
2461 acccggaagt cggaggagac catcaccccg tggaacttcg aggaggtcgt ggacaagggc
2521 gcgtccgcgc agtcgttcat cgagcgcatg accaacttcg acaagaacct cccgaacgag
2581 aaggtcctgc ccaagcactc cctgctctac gagtacttca ccgtgtacaa cgagctgacc
2641 aaggtcaagt acgtgaccga gggcatgcgg aagccggcct tcctgtcggg cgagcagaag
2701 aaggcgatcg tggacctgct cttcaagacc aaccgcaagg tcaccgtgaa gcagctgaag
2761 gaggactact tcaagaagat cgagtgcttc gactccgtcg agatcagcgg cgtggaggac
2821 cgcttcaacg cctccctggg cacctaccac gacctgctca agatcatcaa ggacaaggac
2881 ttcctcgaca acgaggagaa cgaggacatc ctggaggaca tcgtcctcac cctgaccctc
2941 ttcgaggacc gcgagatgat cgaggagcgg ctcaagacct acgcccacct gttcgacgac
3001 aaggtgatga agcagctgaa gcgtcgccgc tacaccggct ggggccgcct ctcccggaag
3061 ctgatcaacg gcatccggga caagcagagc ggcaagacca tcctggactt cctcaagtcc
3121 gacggcttcg ccaaccgcaa cttcatgcag ctcatccacg acgacagcct gaccttcaag
3181 gaggacatcc agaaggccca ggtctcgggc cagggcgaca gcctccacga gcacatcgcc
3241 aacctggcgg gctccccggc gatcaagaag ggcatcctcc agaccgtcaa ggtcgtggac
3301 gagctggtca aggtgatggg ccgccacaag cccgagaaca tcgtgatcga gatggcccgg
3361 gagaaccaga ccacccagaa gggccagaag aactcgcgcg agcggatgaa gcggatcgag
3421 gagggcatca aggagctcgg cagccagatc ctgaaggagc acccggtcga gaacacccag
3481 ctgcagaacg agaagctgta cctctactac ctgcagaacg gccgcgacat gtacgtggac
3541 caggagctcg acatcaaccg gctgtccgac tacgacgtgg accacatcgt gccgcagtcc
3601 ttcctgaagg acgactcgat cgacaacaag gtcctgaccc gctcggacaa gaaccggggc
3661 aagtccgaca acgtgccctc ggaggaggtc gtgaagaaga tgaagaacta ctggcgccag
3721 ctgctcaacg ccaagctcat cacccagcgc aagttcgaca acctgaccaa ggccgagcgg
3781 ggcggcctga gcgagctcga caaggcgggc ttcatcaagc gccagctggt cgagacccgg
3841 cagatcacca agcacgtggc ccagatcctg gactcccgga tgaacaccaa gtacgacgag
3901 aacgacaagc tgatccgcga ggtcaaggtg atcaccctca agagcaagct ggtctccgac
3961 ttccgcaagg acttccagtt ctacaaggtc cgggagatca acaactacca ccacgcccac
4021 gacgcgtacc tgaacgccgt cgtgggcacc gcgctgatca agaagtaccc gaagctggag
4081 tccgagttcg tctacggcga ctacaaggtc tacgacgtgc gcaagatgat cgccaagagc
4141 gagcaggaga tcggcaaggc caccgcgaag tacttcttct actccaacat catgaacttc
4201 ttcaagaccg agatcaccct ggccaacggc gagatccgca agcggcccct gatcgagacc
4261 aacggcgaga ccggcgagat cgtctgggac aagggccgcg acttcgccac cgtccggaag
4321 gtgctgtcga tgccgcaggt caacatcgtg aagaaaaccg aggtgcagac cggcggcttc
4381 agcaaggagt ccatcctccc caagcgcaac agcgacaagc tgatcgcccg gaagaaggac
4441 tgggacccga agaagtacgg cggcttcgac agccccaccg tcgcctactc cgtgctggtc
4501 gtggcgaagg tcgagaaggg caagagcaag aagctgaagt ccgtgaagga gctgctcggc
4561 atcaccatca tggagcgctc ctcgttcgag aagaacccga tcgacttcct ggaggccaag
4621 ggctacaagg aggtcaagaa ggacctcatc atcaagctgc ccaagtacag cctgttcgag
4681 ctggagaacg gccgcaagcg gatgctcgcc tccgcgggcg agctgcagaa gggcaacgag
4741 ctggccctcc cgtcgaagta cgtcaacttc ctgtacctcg cgtcccacta cgagaagctg
4801 aagggctcgc ccgaggacaa cgagcagaag cagctcttcg tggagcagca caagcactac
4861 ctggacgaga tcatcgagca gatcagcgag ttcagcaagc gcgtcatcct ggccgacgcg
4921 aacctcgaca aggtgctgtc cgcctacaac aagcaccgcg acaagccgat ccgggagcag
4981 gcggagaaca tcatccacct gttcaccctc accaacctgg gcgcccccgc cgcgttcaag
5041 tacttcgaca ccaccatcga ccgcaagcgg tacacctcca ccaaggaggt cctcgacgcg
5101 accctgatcc accagagcat caccggcctg tacgagaccc gcatcgacct gtcccagctc
5161 ggcggcgact ccggcggctc caccaacctg tccgacatca tcgagaagga gaccggcaag
5221 cagctggtca tccaggagtc catcctgatg ctgccggagg aagtcgagga agtcatcggc
5281 aacaagcccg agtccgacat cctggtccac accgcctacg acgagtccac cgacgagaac
5341 gtcatgctgc tgacctcgga cgcccccgag tacaagccct gggccctggt catccaggac
5401 tccaacggcg agaacaagat caagatgctg tgatctagag gatccccggg taccgagctc
5461 ggattcccca gatctaaagt tttgtcgtct ttccagacgt tagtaaatga attttctgta
5521 tgaggttttg ctaaacaact ttcaacagtt tcagcggagt gagaatagaa aggaacaact
5581 aaaggaattg cgaataataa ttttttcacg ttgaaaatct ccaaaaaaaa aggctccaaa
5641 aggagccttt aattgtatcg gtttatcagc ttgctttcga ggtgaatttc ttaaacagct
5701 tgataccgat agttgcgccg acaatgacaa caaccatcgc ccacgcataa ccgatatatt
5761 cgactagtct gttggctact ctatcattga tagtggtagg atccctatca gtgatagaga
5821 taagtcttct cagccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg
5881 ggaaggggga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
5941 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
6001 ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttgggtaccg ggccccccct
6061 cgaggtcctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat
6121 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
6181 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
6241 cgttgcgctc actgcccgct tccaccggtg aagacatgtt ttagagctag aaatagcaag
6301 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgctttttac
6361 tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatc tagaggccag
6421 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
6481 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
6541 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
6601 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
6661 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg acccccccgt
6721 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
6781 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
6841 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
6901 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
6961 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
7021 cagaaaaaaa ggatctcaag cggccgcatc ccattatgct ttggcagttt attcttgaca
7081 tgtagtgagg gggctggtat aatcacatac ggccgataaa gcaagcatat aatattgcgt
7141 ttcatcttta gaagcgaatt tcgccaatat tataattatc aaagagaggg gtggcaaacg
7201 gtatttggca ttattaggtt aaaaaatgta gaaggagagt gaaacccatg aactttaata
7261 aaattgattt agacaattgg aagagaaaag agatatttaa tcattatttg aaccaacaaa
7321 cgacttttag tataaccaca gaaattgata ttagtgtttt ataccgaaac ataaaacaag
7381 aaggatataa attttaccct gcatttattt tcttagtgac aagggtgata aactcaaata
7441 cagcttttag aactggttac aatagcgacg gagagttagg ttattgggat aagttagagc
7501 cactttatac aatttttgat ggtgtatcta aaacattctc tggtatttgg actcctgtaa
7561 agaatgactt caaagagttt tatgatttat acctttctga tgtagagaaa tataatggtt
7621 cggggaaatt gtttcccaaa acacctatac ctgaaaatgc tttttctctt tctattattc
7681 catggacttc atttactggg tttaacttaa atatcaataa taatagtaat taccttctac
7741 ccattattac agcaggaaaa ttcattaata aaggtaattc aatatattta ccgctatctt
7801 tacaggtaca tcattctgtt tgtgatggtt atcatgcagg attgtttatg aactctattc
7861 aggaattgtc agataggcct aatgactggc ttttataata tgagataatg ccgactgtac
7921 tttttacagt cggttttcta atgtcactaa cctgccccgt tagttgatgc atgatcatca
7981 taattctgtc tcattatcta gagtcgccat tggggaagaa gagcaggatt tgcctgcggt
8041 tgaccgagac ggagtgggcg acgatgaacc ggatggcgtt cgcgtgcggt cagatggata
8101 tctccatgga cacccacatg gatatctcca tggacaccca catggatatc tccatggaca
8161 cccacatgga tatctccatc aaatcgaagg taagcaagct agacggttca tgtacatgaa
8221 ccgaacgcag cacctccctc gccttgcggc gctggagcaa caacccgagt gccgaaggcc
8281 gatggaagag gaaaaagccg caaggcgagg gaggtgctgc gttcgcgttt gggacaacgt
8341 ttagcgtcag tcgaacagtg gaatctcgcc gggtagatgc ttcgatttct ccgccgcctc
8401 gtcctcgcgc tccttgcgag ccctgtagca gcgttgacgc acggcaccct ggctcatgtt
8461 gaaatgctcg gcggtctgcg gccatgtatg gccctcgtca tcgtgatatg agcggactgc
8521 ttcgcgcata gcggcttgtt catcaatcca atcctcgcgc ttcattgcca cccagctctg
8581 aatggtgcgc gtggagacgc cgtatgcttc agctaattca cgtgcagaca ccgggcgctt
8641 cttcctcaaa gtcgtcctta ccattccagt gcctccttca tgacctgctg atatttgttc
8701 tcaccgtgtt tgtgtcctcg tgcggattgg atggcgatga atgtggctgc gttggcaatg
8761 gcaccgtcga gccacatgcg gctgcgggtg acgatccatt tgtggatgct cttggcgatg
8821 tcctcgacct cacgcgcgtg cagcggatcg gggaacagcg agacgttgag ttcgtggcag
8881 gtgcggcgca cgtataggcg caatgcctcg ctgctggcct ccgaacggtc gggcagccgc
8941 cgcacgtagc ggtaggcgag cgtgcgggcc ttgtcgaaga gcgtgcagtt gcgtcccagc
9001 ccctgcgtgc gggcgcgctt ggtgcgcttc cagctgacgg gcggcatgtc cccgtgttcc
9061 tggagcgact gcacgagctg ttccaggtcg taggtgtccc actcgatgat gtccgacgcc
9121 cacgccggat gctcggggtt cttcatcaga agtcccgaat agcccatgtc cccgtcgcag
9181 gagcggcgca gtccctccgt ggtggcgtgc aggagcttca acggcttgag acgcgccaga
9241 tcggttctgg gcaccggaaa ggtgagcacc cagcccgcgt gagcgtgccc gttggccggg
9301 ttctccgcga tccagttcgg ccgcatgccc tcgtgctccc acagggccat cgccttggcg
9361 ttcgcgtcgt cgatgtccac cacgatcgtg ttcaccagcg cggcggggtt cgcctcgatg
9421 tgcttgcatt tgagcgcgtc ggcgcggctc ctgcggtgca gctgggagta gtcgtcgtcg
9481 cagcacagcg ggcgttcggg cagccaccag ccttcgaacg tgcgcgacag ctcaagcgtc
9541 ggctgcgaat actcatcgga catggctaca gcatacatcg tatttgtaca gtgaacaaat
9601 acgcgggaac caagtcagat atttcagccc agcgaagcgt cggctgaacc cagcccagca
9661 catcctgcac gaaaagccgc tggacggccc tcagccgctt ctccacgccg ctcacgtatg
9721 cgcgagcctc tctgacctgt tcggccgcgt ggaggccact cacggacgtt tctgagccgt
9781 ctgagccgtc ggcgtccgct gcgcgttccg caggctgagg c
//
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cBEST4质粒载体应用举例
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