| 质粒名称: | pST1374-GCN4-D10A |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 113022 |
| 存储实验室: | Xingxu Huang |
|
载体骨架基本信息 |
|
| 载体名称: | pST1374 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4968 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | GCN4-D10A,2xNLS-10xGCN4-NLS-D10A-NLS |
| 物种来源: | S. cerevisiae (budding yeast); S. pyogenes |
| 插入大小: | 5634 bp |
| 基因突变: | D10A |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | -- |
| 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 | CMV-fwd |
| 3' 测序引物 | BGH-rev |
|
文献引用方法 |
|
| 材料和方法部分: | pST1374-GCN4-D10A was a gift from Xingxu Huang (Addgene plasmid # 113022 ; http://n2t.net/addgene:113022 ; RRID:Addgene_113022) |
| 参考文献部分: | BE-PLUS: a new base editing tool with broadened editing window and enhanced fidelity. Jiang W, Feng S, Huang S, Yu W, Li G, Yang G, Liu Y, Zhang Y, Zhang L, Hou Y, Chen J, Chen J, Huang X. Cell Res. 2018 Jun 6. pii: 10.1038/s41422-018-0052-4. doi: 10.1038/s41422-018-0052-4. 10.1038/s41422-018-0052-4 [pii] PubMed 29875396 |
买家导航
pST1374-GCN4-D10A载体质粒基本信息
pST1374-GCN4-D10A质粒图谱载体图谱和pST1374-GCN4-D10A载体序列质粒序列多克隆位点信息
pST1374-GCN4-D10A质粒载体简介
pST1374-GCN4-D10A质粒序列载体序列
LOCUS pST1374-GCN4-D10A 10601 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expresses GCN4-D10A in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pST1374-GCN4-D10A
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10601)
AUTHORS Jiang W, Feng S, Huang S, Yu W, Li G, Yang G, Liu Y, Zhang Y, Zhang
L, Hou Y, Chen J, Chen J, Huang X
TITLE BE-PLUS: a new base editing tool with broadened editing window and
enhanced fidelity.
JOURNAL Cell Res. 2018 Jun 6. pii: 10.1038/s41422-018-0052-4. doi:
10.1038/s41422-018-0052-4.
PUBMED 29875396
REFERENCE 2 (bases 1 to 10601)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..10601
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 629..648
/label=T7
/note="T7 promoter, forward primer"
promoter 629..647
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 676..696
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 703..723
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 760..816
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 898..954
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1036..1092
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1174..1230
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1312..1368
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1450..1506
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1588..1644
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1726..1782
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1864..1920
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 2002..2058
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 2098..2118
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 2125..6225
/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 6256..6276
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 6283..6300
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(6323..6340)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 6329..6553
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 6599..7027
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6686..6705)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6896..6917
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(7036..7056)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 7041..7370
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 7221..7356
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 7283..7302
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 7418..7465
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 7484..7882
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/label=BSD
/note="confers resistance to blasticidin"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
polyA_signal 8040..8161
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(8077..8096)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 8131..8150
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(8210..8226)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8210..8226)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(8223..8245)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 8234..8250
/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(8258..8288)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8303..8324
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(8441..8458)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(8612..9200)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(8692..8711)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(9371..10231)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 9994..10013
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(10232..10336)
/gene="bla"
/label=AmpR promoter
primer_bind complement(10411..10430)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
661 gctagcacca tgggacccaa gaaaaaacgc aaggtggaag atcctaagaa aaagcggaaa
721 gtggacggca ttggtagtgg gagcaacggc agcagcggag aggaactgct gagcaagaac
781 taccacctgg aaaacgaggt ggccagactg aagaaaggct ctggctctgg cggaagcggt
841 tctggcggat caggatctgg aagtggcggc tctggaagcg gaggttctgg atctggcgaa
901 gaactgctgt ctaagaatta tcacctcgag aacgaagtgg ctcggctcaa gaaaggcagt
961 ggcagcggag gaagtggatc cggcggtagt ggtagtggaa gcggcggatc cggctcaggc
1021 ggatctggtt caggggaaga actcctctcc aaaaactacc atctcgagaa tgaggtcgcc
1081 cgcctgaaaa aaggatcagg ttccggtggt tctggcagcg gtggttcagg ctcaggcagc
1141 ggcggtagcg gtagcggtgg aagcggaagc ggagaagaac ttctcagcaa aaattaccac
1201 ttggagaatg aagttgcaag actcaaaaaa ggttccggca gtggcggcag cggcagcggc
1261 ggatctggta gtggatctgg tggcagtggt tcaggcggaa gtggtagcgg agaggaattg
1321 ctctcaaaga actatcattt ggagaacgag gttgcacgct tgaagaaagg cagcggatca
1381 ggcggatctg gcagcggtgg atctggttct ggatccggcg gctccggtag tggtggaagt
1441 ggctctgggg aagaattgct tagcaagaat tatcatcttg aaaatgaggt tgccaggctt
1501 aaaaaaggca gtggctccgg cggatccgga agcggaggca gcggatctgg atctggtggt
1561 tcaggatctg gcggttctgg tagcggggaa gaactgttga gtaaaaacta tcaccttgag
1621 aacgaggtcg caaggttgaa aaaaggatcc ggctctggcg gctccggaag tggcggatct
1681 ggctccggta gcggaggatc aggatccggc ggaagcggat caggcgagga actgctttcc
1741 aaaaattacc accttgaaaa cgaagtcgcc cgcctcaaga aaggttctgg cagcggaggc
1801 tctggcagtg gtggtagcgg aagtggaagt ggtggcagtg gtagcggtgg atctggaagc
1861 ggcgaggaac tcctgtcaaa gaattaccat ctcgaaaacg aggtcgcaag gctcaagaaa
1921 ggctcaggat caggcggctc tggatccggc ggttctggtt ccggctcagg tggaagtgga
1981 tctggcggct caggttccgg cgaagaattg ctttccaaga actaccattt ggaaaatgaa
2041 gtcgctcgtt tgaagaaagg ttcaggctcc ggccagcggc cgcaaggtgg aggtggaccc
2101 aagaagaagc gcaaggtgtc tagagacaag aaatactcta ttggactggc tatcgggaca
2161 aactccgttg gctgggccgt cataaccgac gagtataagg tgccaagcaa gaaattcaag
2221 gtgctgggta atactgaccg ccattcaatc aagaagaacc tgatcggagc actcctcttc
2281 gactccggtg aaaccgctga agctactcgg ctgaagcgga ccgcaaggcg gagatacacc
2341 cgccgcaaga atcggatatg ttatctgcaa gagatcttta gcaacgaaat ggctaaggtg
2401 gacgactcct tctttcaccg cctggaagag agctttctgg tggaggagga taagaaacac
2461 gagaggcacc ctatattcgg aaatatcgtg gatgaggtgg cttaccatga aaagtatcct
2521 acaatctacc atctgaggaa gaagctggtg gacagcaccg ataaagcaga cctgaggctc
2581 atctatctgg ccctggctca tatgataaag tttagaggac actttctgat cgagggcgac
2641 ctgaatcccg ataattccga tgtggataaa ctcttcattc aactggtgca gacatataac
2701 caactgttcg aggagaatcc cataaacgct tctggtgtgg atgccaaggc tattctgtcc
2761 gctcggctgt ccaagtcacg cagactggag aatctgattg cccaactgcc aggagaaaag
2821 aagaacggcc tgtttgggaa cctcatcgcc ctgagcctgg gcctgacacc taacttcaag
2881 tccaattttg atctggccga agatgctaaa ctccagctct ccaaggacac ctatgacgat
2941 gatctggaca acctgctcgc acagataggc gaccagtacg ccgatctctt tctggctgct
3001 aagaatctct ccgacgccat tctgctgagc gacatactcc gggtcaacac tgagatcacc
3061 aaagcacctc tgagcgcctc catgataaaa cgctatgatg aacaccatca agacctgact
3121 ctgctcaaag ccctcgtgag gcaacagctg ccagagaagt acaaagagat attcttcgac
3181 cagagcaaga atggatatgc cggatacatc gatggcggag catcacagga agaattttac
3241 aagttcatca aaccaatcct cgagaagatg gacggtactg aagagctgct ggtgaagctg
3301 aacagggagg acctgctgag gaagcagagg acctttgata atggctccat tccacatcag
3361 atacacctgg gagagctgca tgcaatcctc cgcaggcagg aggatttcta tcctttcctg
3421 aaggataacc gggagaagat agagaagatc ctgaccttca ggatccctta ttacgtcggc
3481 cctctggcta gaggcaactc ccgcttcgct tggatgacca ggaaatctga ggagacaatt
3541 actccttgga acttcgaaga ggtcgtggat aagggcgcaa gcgcccagtc attcatcgaa
3601 cggatgacca atttcgataa gaacctgccc aacgagaagg tcctgcccaa acattcactc
3661 ctgtacgagt atttcaccgt ctataacgag ctgactaaag tgaagtacgt gaccgagggc
3721 atgaggaagc ctgccttcct gtccggagag cagaagaagg ctatcgttga tctgctcttc
3781 aagactaata gaaaggtgac agtgaagcag ctcaaggagg attactttaa gaagatcgaa
3841 tgctttgact cagtggaaat ctctggcgtg gaggaccgct ttaatgccag cctgggcact
3901 taccatgatc tgctgaagat aatcaaagac aaagatttcc tcgataatga ggagaacgag
3961 gacatcctgg aagatatcgt gctgaccctg actctgttcg aggatagaga gatgatcgaa
4021 gagcgcctga agacctatgc ccatctgttt gacgataaag tcatgaaaca gctcaagcgg
4081 cggcgctaca ctgggtgggg tagactctcc aggaaactca taaacggcat ccgcgacaaa
4141 cagagcggaa agaccatcct ggatttcctg aaatccgacg gattcgctaa caggaacttc
4201 atgcaactga ttcacgatga ctctctgaca tttaaagagg acatccagaa ggcacaggtg
4261 agcggtcaag gcgacagcct gcacgagcac atcgccaacc tcgctggatc acccgccata
4321 aagaagggaa tactgcagac agtcaaggtc gtggacgaac tcgtcaaagt gatgggtcgg
4381 cacaagccag agaatatcgt tatcgaaatg gcaagggaga accaaaccac ccagaagggc
4441 cagaagaact ctcgggaacg gatgaaaaga atcgaagagg gaattaagga gctgggatct
4501 cagatactga aggagcaccc tgtggagaat acacagctcc agaacgagaa actctacctg
4561 tactacctcc agaacgggcg ggacatgtac gttgaccagg aactcgacat caaccggctg
4621 tccgattatg acgtggacca tattgttcca cagtccttcc tcaaagatga ctccattgac
4681 aacaaggtgc tgaccagatc cgataagaat cgcggtaagt ctgacaatgt tccatcagaa
4741 gaggtggtca agaagatgaa gaattactgg cggcagctcc tcaacgccaa actgatcacc
4801 cagcggaagt ttgacaatct gactaaggca gaaagaggag gtctgagcga actcgacaag
4861 gccggcttta ttaagaggca actggtcgaa acacgccaga ttaccaaaca cgtggcacaa
4921 atcctcgact ctaggatgaa cactaagtac gatgagaacg ataagctgat cagggaagtg
4981 aaagtgataa ctctgaagag caagctggtg tctgacttcc ggaaggactt tcaattctac
5041 aaagttcgcg aaataaacaa ttaccatcat gctcacgatg cctatctcaa tgctgtcgtt
5101 ggcaccgccc tgatcaagaa ataccctaaa ctggagtctg agttcgtgta cggtgactat
5161 aaagtctacg atgtgaggaa gatgatagca aagtctgagc aagagattgg caaagccacc
5221 gccaagtact tcttctactc taatatcatg aatttcttta agactgagat aaccctggct
5281 aacggcgaaa tccggaagcg cccactgatc gaaacaaacg gagaaacagg agaaatcgtg
5341 tgggataaag gcagggactt cgcaactgtg cggaaggtgc tgtccatgcc acaagtcaat
5401 atcgtgaaga agaccgaagt gcagaccggc ggattctcaa aggagagcat cctgccaaag
5461 cggaactctg acaagctgat cgccaggaag aaagattggg acccaaagaa gtatggcggt
5521 ttcgattccc ctacagtggc ttattccgtt ctggtcgtgg caaaagtgga gaaaggcaag
5581 tccaagaaac tcaagtctgt taaggagctg ctcggaatta ctattatgga gagatccagc
5641 ttcgagaaga atccaatcga tttcctggaa gctaagggct ataaagaagt gaagaaagat
5701 ctcatcatca aactgcccaa gtactctctc tttgagctgg agaatggtag gaagcggatg
5761 ctggcctccg ccggagagct gcagaaagga aacgagctgg ctctgccctc caaatacgtg
5821 aacttcctgt atctggcctc ccactacgag aaactcaaag gtagccctga agacaatgag
5881 cagaagcaac tctttgttga gcaacataaa cactacctgg acgaaatcat tgaacagatt
5941 agcgagttca gcaagcgggt tattctggcc gatgcaaacc tcgataaagt gctgagcgca
6001 tataataagc acagggacaa gccaattcgc gaacaagcag agaatattat ccacctcttt
6061 actctgacta atctgggcgc tcctgctgcc ttcaagtatt tcgatacaac tattgacagg
6121 aagcggtaca cctctaccaa agaagttctc gatgccaccc tgatacacca gtcaattacc
6181 ggactgtacg agactcgcat cgacctgtct cagctcggcg gcgacggttc tgaattcagc
6241 ctgggcagcg gctcccccaa gaagaagcgc aaggtgaccg gtcatcatca ccatcaccat
6301 tgagtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
6361 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
6421 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
6481 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
6541 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
6601 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
6661 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
6721 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
6781 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
6841 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
6901 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
6961 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
7021 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
7081 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
7141 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
7201 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
7261 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
7321 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
7381 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata
7441 gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca agcctttgtc
7501 tcaagaagaa tccaccctca ttgaaagagc aacggctaca atcaacagca tccccatctc
7561 tgaagactac agcgtcgcca gcgcagctct ctctagcgac ggccgcatct tcactggtgt
7621 caatgtatat cattttactg ggggaccttg tgcagaactc gtggtgctgg gcactgctgc
7681 tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat
7741 cttgagcccc tgcggacggt gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc
7801 catagtgaag gacagtgatg gacagccgac ggcagttggg attcgtgaat tgctgccctc
7861 tggttatgtg tgggagggct aagcacttcg tggccgagga gcaggactga cacgtgctac
7921 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
7981 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
8041 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
8101 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
8161 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
8221 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
8281 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
8341 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
8401 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
8461 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
8521 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
8581 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
8641 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
8701 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
8761 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
8821 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
8881 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
8941 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
9001 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
9061 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
9121 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
9181 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
9241 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
9301 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
9361 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
9421 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
9481 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
9541 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
9601 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
9661 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
9721 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
9781 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
9841 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
9901 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
9961 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
10021 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
10081 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
10141 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
10201 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
10261 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
10321 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
10381 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
10441 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
10501 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
10561 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt t
//
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pST1374-GCN4-D10A质粒载体应用举例
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