| 质粒名称: | SuperFiCas9-tRNA-gRNA-all-in-One |
|---|---|
| 出品公司: | Wekwikgene |
| 目录编号: | 0002157 |
| 产品用途: | Mammalian expression of Cas9 and sgRNA scaffold |
| 存储实验室: | Westlake Genetech |
|
载体骨架基本信息 |
|
| 载体名称: | LV-SFCas9 |
| 表达系统: | 慢病毒，哺乳动物表达 |
| 启动子: |
CMV, U6
|
| 拷贝数: | 高拷贝 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | Stbl3 |
|
插入基因信息 |
|
| 插入基因名称: | superFiCas9, GFP |
| 物种来源: | H.sapiens |
| 密码子优化: | 未优化 |
|
文献引用方法 |
|
| 参考文献部分: |
Spacer Extension Restores Cleavage Activity by Enhancing PAM-Distal Interactions in High-Fidelity Cas9
Rong Zheng、Zhike Lu、Rongwei Wei et al.
Nature Structural & Molecular Biology
|
买家导航
SuperFiCas9-tRNA-gRNA-all-in-One载体质粒基本信息
SuperFiCas9-tRNA-gRNA-all-in-One质粒图谱载体图谱和SuperFiCas9-tRNA-gRNA-all-in-One载体序列质粒序列多克隆位点信息
SuperFiCas9-tRNA-gRNA-all-in-One质粒载体简介
SuperFiCas9-tRNA-gRNA-all-in-One质粒序列载体序列
LOCUS SuperFiCas9-tRNA-gRNA-all-in-One 14969 bp DNA circular SYN 06-NOV-2025
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14969)
AUTHORS Westlake University
TITLE Direct Submission
JOURNAL Exported Apr 24, 2026 from SnapGene 6.0.2
https://www.biofeng.com
FEATURES Location/Qualifiers
source 1..14969
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
enhancer 77..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
misc_feature 136..142
/label=cAMP/PKA
misc_feature complement(137..143)
/label=cAMP/PKA
misc_feature 177..186
/label=NFkBBinding
misc_feature 189..195
/label=cAMP/PKA
misc_feature complement(190..196)
/label=cAMP/PKA
misc_feature 272..278
/label=cAMP/PKA
misc_feature complement(273..279)
/label=cAMP/PKA
misc_feature 328..337
/label=NFkBBinding
promoter 381..584
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 382..580
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
misc_feature 425..431
/label=AP1
/note="AP1BindingSite"
misc_feature 458..464
/label=cAMP/PKA
misc_feature complement(459..465)
/label=cAMP/PKA
misc_feature 496..505
/label=NFkBBinding
primer_bind 535..555
/label=CMV-F
primer_bind 547..566
/label=pIRES2-EGFP-P5
misc_feature 570..576
/label=CMV-TATA
LTR 598..778
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 598..778
/label=5LTR-zhike
misc_feature 599..619
/label=5LTRRstart(+1mRNA)-Lenti
misc_feature 600..695
/label=HXB2-R
misc_feature 600..619
/label=5LTRRstart(+1mRNA)-Lenti
misc_feature 620..657
/label=TARbulge-loop-Lenti
misc_feature 671..676
/label=PolyAMotif
misc_feature 696..778
/label=HXB2-U5
misc_feature 696..778
/label=U5-Lenti
misc_feature complement(780..797)
/label=PBS(tRNALys3Human)
misc_feature 799..817
/label=InterferonResponseElement
misc_feature 825..950
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 888..911
/label=Intron5SplicingSite1
CDS 934..977
/codon_start=1
/label=HXB2-GagStart
/translation="MGARASVLSGGELD"
misc_feature complement(1167..1173)
/label=CMV-TATA
misc_feature 1168..1174
/label=CMV-TATA
misc_feature 1230..1235
/label=OriginTSS
misc_feature complement(1345..1351)
/label=CMV-TATA
misc_feature 1346..1352
/label=CMV-TATA
misc_feature 1443..1676
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 1677..2079
/label=RRE-con
misc_feature 1744..1749
/label=PolyAMotif
CDS 1861..1905
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 1939..1945
/label=CMV-TATA
misc_feature 1979..1986
/label=Stem-tetraV1
misc_feature complement(2031..2052)
/label=Intron3SplicingSite2
misc_feature 2046..2053
/label=Stem-tetraV2
misc_feature 2177..2202
/label=cPPT-con
misc_feature 2203..2320
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
misc_feature 2207..2221
/label=PPT(HIV1)
misc_feature 2321..2356
/label=cPPT-con2
misc_feature complement(2325..2330)
/label=PolyAMotif
primer_bind 2340..2390
/label=Primer 2
primer_bind complement(2342..2356)
/label=gRNA-R
promoter 2366..2614
/label=U6-zhike
primer_bind 2366..2386
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 2537..2556
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_feature 2575..3558
/label=W41G10-2_U6_TSS20241111-0571-04927-01_F11
misc_feature 2616..2687
/label=tRNA
misc_feature 2710..2717
/label=Stem-tetraV2
misc_feature 2718..2726
/label=Stem-tetraV1
misc_feature 2718..2723
/label=Stem-tetraV2
misc_feature complement(2733..2741)
/label=Stem-tetraV1
misc_feature complement(2736..2741)
/label=Stem-tetraV2
misc_feature 2742..2769
/label=Stem1
misc_feature 2770..2775
/label=Stem2V2
misc_feature complement(2782..2787)
/label=Stem2V2
misc_feature 2788..2803
/label=Stem3
primer_bind complement(2804..2847)
/label=Primer 1
misc_feature 2804..2811
/label=Termini
misc_feature 2804..2810
/label=Termini
misc_feature 2826..3499
/label=A2UCOE
primer_bind complement(3432..3441)
/label=Mit-7F
promoter 3581..4759
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
primer_bind complement(3699..3718)
/label=EF1a-R
misc_feature 3749..3755
/label=CMV-TATA
misc_feature 3779..3811
/label=TSS-EF1a
intron 3812..4750
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
misc_feature complement(3918..3923)
/label=OriginTSS
misc_feature complement(4447..4453)
/label=AP1
/note="AP1BindingSite"
primer_bind 4707..4727
/label=PEF-F(EF-1a-F)
CDS 4781..4834
/codon_start=1
/label=NLS(BE4Max1)
/translation="KRTADGSEFESPKKKRKV"
CDS 4814..4834
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 4835..8935
/codon_start=1
/product="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
/label=Cas9
/note="generates RNA-guided double strand breaks in DNA"
/translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVDGDDKVDDDDKMIAKSEDEIGDATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
misc_feature complement(5117..5122)
/label=OriginTSS
misc_feature 6004..6009
/label=OriginTSS
misc_feature complement(6299..6304)
/label=OriginTSS
primer_bind complement(6817..6827)
/label=pC1300-F
misc_feature 6823..6828
/label=OriginTSS
misc_feature 7258..7263
/label=OriginTSS
misc_feature 7349..7351
/label=840
mat_peptide 7859..7861
/label=Y1010D
mat_peptide 7868..7870
/label=Y1013D
mat_peptide 7878..7879
/label=Y1016D
mat_peptide 7883..7885
/label=V1018D
misc_feature 7886..7888
/label=R1019D
misc_feature 7910..7912
/label=Q1027D
misc_feature 7922..7924
/label=K1031D
misc_feature complement(8348..8353)
/label=OriginTSS
primer_bind complement(8438..8458)
/label=pEGFP-N3(EGFP-N)
misc_feature 8560..8565
/label=OriginTSS
CDS 8948..8998
/codon_start=1
/label=NLS(BE4Max2)
/translation="KRTADGSEFEPKKKRKV"
CDS 8948..8969
/codon_start=1
/label=NLS(BE4Max2)
/translation="KRTADGS"
CDS 8978..8998
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 8999..9022
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 9032..9088
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS 9092..9805
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
primer_bind complement(9134..9155)
/label=pEGFP-N3(EGFP-N)
primer_bind complement(9293..9312)
/label=GFP-R-jian
primer_bind complement(9296..9315)
/label=SP1305G-R
primer_bind 9742..9763
/label=pEGFP-C5（EGFP-C)
misc_feature 9824..10412
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(9877..9897)
/label=WPRE-R
CDS complement(10295..10306)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
misc_feature 10418..10483
/label=3LTRcon
misc_feature 10467..10491
/label=PolypurineTract-Lenti
misc_feature 10467..10481
/label=PPT(HIV1)
LTR 10484..10717
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
LTR 10537..10717
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 10537..10717
/label=5LTR-zhike
misc_feature 10539..10558
/label=5LTRRstart(+1mRNA)-Lenti
misc_feature 10559..10596
/label=TARbulge-loop-Lenti
misc_feature 10610..10615
/label=PolyAMotif
misc_feature 10635..10717
/label=U5-Lenti
primer_bind complement(10743..10760)
/label=BGH
polyA_signal 10749..10973
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
polyA_signal 10754..10973
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
misc_feature 10839..10844
/label=PolyAMotif
rep_origin 11019..11447
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(11456..11476)
/label=PBABE-R
promoter 11461..11790
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
misc_feature complement(11479..11488)
/label=NFkBBinding
primer_bind complement(11524..11543)
/label=pLXSN-R
misc_feature complement(11551..11560)
/label=NFkBBinding
primer_bind complement(11596..11615)
/label=pLXSN-R
rep_origin 11641..11776
/label=SV40 ori
/note="SV40 origin of replication"
misc_feature 11696..11703
/label=Termini
misc_feature complement(11701..11706)
/label=PolyAMotif
primer_bind 11703..11722
/label=SV40pro-F
promoter 11838..11885
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 11904..12278
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=BleoR
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
polyA_signal 12408..12537
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
polyA_signal 12408..12529
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
misc_feature complement(12414..12419)
/label=PolyAMotif
misc_feature 12439..12444
/label=PolyAMotif
primer_bind complement(12445..12464)
/label=SV40-PArev
misc_feature 12468..12473
/label=PolyAMotif
primer_bind 12499..12518
/label=EBV-R
primer_bind 12507..12526
/label=pCMV-R
primer_bind complement(12578..12594)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(12596..12615)
/label=pQE30-F
protein_bind 12602..12618
/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(12626..12656)
/label=lac promoter
/note="promoter for the E. coli lac operon"
primer_bind complement(12634..12653)
/label=pC1300-F
primer_bind complement(12643..12662)
/label=PCAMBIA1301-F
protein_bind 12671..12692
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
misc_feature complement(12741..12763)
/label=Test-500-F
misc_feature 12921..13637
/label=OriPromotor
rep_origin complement(12980..13568)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 12987..12999
/label=Origin-GC
misc_feature 13042..13058
/label=OriginTSS
misc_feature 13057..13064
/label=Origin-TATA
misc_feature 13084..13100
/label=OriginTSS
misc_feature 13084..13089
/label=OriginTSS
misc_feature complement(13106..13126)
/label=5SplicingSite
misc_feature complement(13271..13289)
/label=OriginTSS
CDS complement(13739..14599)
/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"
misc_feature 13855..13871
/label=Test-500-R2
misc_feature 13913..13918
/label=PolyAMotif
misc_feature 13998..14003
/label=OriginTSS
misc_feature complement(14351..14373)
/label=Test-500-F2
primer_bind 14362..14381
/label=AMP-R
promoter complement(14600..14704)
/gene="bla"
/label=AmpR promoter
primer_bind 14635..14656
/label=163-F
misc_feature 14682..14687
/label=PolyAMotif
misc_feature 14729..14735
/label=cAMP/PKA
misc_feature complement(14778..14798)
/label=Test-500-R
primer_bind 14926..14938
/label=Mit-24R
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 cgcgttttgc ctgtactggg
601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
2341 cagagatcca gtttggttaa ttagagaggg cctatttccc atgattcctt catatttgca
2401 tatacgatac aaggctgtta gagagataat tagaattaat ttgactgtaa acacaaagat
2461 attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa
2521 attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt
2581 ggctttatat atcttgtgga aaggacgaaa caccgggttc catggtgtaa tggttagcac
2641 tctggactct gaatccagcg atccgagttc aaatctcggt ggaacctatc ggcaggtgca
2701 cctgctacgg tttgagagct atgctgcggc cgcagcatag caagttcaaa taaggctagt
2761 ccgttatcaa cttggcggcc gccaagtggc accgagtcgg tgcttttttt tgcggccgcg
2821 gccgccgcgt gtggcatctg aagcaccacc agcgagcgag agctagagag aaggaaagcc
2881 accgacttca ccgcctccga gctgctccgg gtcgcgggtc tgcagcgtct ccggccctcc
2941 gcgcctacag ctcaagccac atccgaaggg ggagggagcc gggagctgcg cgcggggccg
3001 ccggggggag gggtggcacc gcccacgccg ggcggccacg aagggcgggg cagcgggcgc
3061 gcgcccggcg gggggagggg ccgcgcgccg cgcccgctgg gaattggggc cctaggggga
3121 gggcggaggc gccgacgacc gcggcactta ccgttcgcgg cgtggcgccc ggtggtcccc
3181 aaggggaggg aagggggagg cggggcgagg acagtgaccg gagtctcctc agcggtggct
3241 tttctgcttg gcagcctcag cggctggcgc caaaaccgga ctccgcccac ttcctcgccc
3301 ctgcggtgcg agggtgtgga atcctccaga cgctggggga gggggagttg ggagcttaaa
3361 aactagtacc cctttgggac cactttcagc agcgaactct cctgtacacc aggggtcagt
3421 tccacagacg cgggccaggg gtgggtcatt gcggcgtgaa caataatttg actagaagtt
3481 gattcgggtg tttccggaac tgcaaagatg gataaagttt taaacagaga ggaatctttg
3541 cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg cccgtcagtg
3601 ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac
3661 cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg
3721 cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct
3781 ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt cccgcgggcc
3841 tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct ggctgcagta
3901 cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc
3961 ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg
4021 cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt ctctagccat
4081 ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc
4141 gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg acggggcccg
4201 tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg
4261 acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat
4321 cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag cggaaagatg
4381 gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct cgggagagcg
4441 ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg
4501 tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga gcttttggag
4561 tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg
4621 ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttgccct
4681 ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct
4741 tccatttcag gtgtcgtgac gtacgagagc cgccaccatg aaacggacag ccgacggaag
4801 cgagttcgag tcaccaaaga agaagcggaa agtcgacaag aagtacagca tcggcctgga
4861 catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa
4921 gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc
4981 cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag
5041 aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat
5101 ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga
5161 taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga
5221 gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga
5281 cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat
5341 cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca
5401 gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc
5461 catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc
5521 cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc
5581 caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac
5641 ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt
5701 tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac
5761 cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca
5821 ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat
5881 tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga
5941 agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct
6001 cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat
6061 cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta
6121 cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta
6181 ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga
6241 ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag
6301 cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa
6361 gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt
6421 gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga
6481 cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa
6541 gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc
6601 cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga
6661 ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga
6721 gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca
6781 gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat
6841 ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa
6901 cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa
6961 agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag
7021 ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt
7081 gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac
7141 ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga
7201 gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa
7261 gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat
7321 caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc tgaaggacga
7381 ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt
7441 gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa
7501 gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga
7561 actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca
7621 cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat
7681 ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt
7741 ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa
7801 cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgga
7861 cggcgacgac aaggtggacg acgacgataa gatgatcgcc aagagcgagg acgaaatcgg
7921 cgacgctacc gccaagtact tcttctacag caacatcatg aactttttca agaccgagat
7981 taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg
8041 ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc
8101 ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat
8161 cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa
8221 gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga
8281 aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga
8341 aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt
8401 gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg
8461 gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc
8521 caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga
8581 ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat
8641 cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt
8701 gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat
8761 ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac
8821 catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca
8881 gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag gtgactctgg
8941 cggctcaaaa agaaccgccg acggcagcga attcgagccc aagaagaaga ggaaagtcga
9001 ttacaaagac gatgacgata agggatccgg cgcaacaaac ttctctctgc tgaaacaagc
9061 cggagatgtc gaagagaatc ctggaccgat ggtgtctaag ggcgaagagc tgttcaccgg
9121 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
9181 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
9241 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
9301 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
9361 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
9421 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
9481 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
9541 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
9601 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
9661 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
9721 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
9781 tctcggcatg gacgagctgt acaagtaaac gcgttaagtc gacaatcaac ctctggatta
9841 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
9901 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
9961 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
10021 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
10081 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
10141 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
10201 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
10261 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
10321 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
10381 gagtcggatc tccctttggg ccgcctcccc gcgtcgactt taagaccaat gacttacaag
10441 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
10501 tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
10561 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
10621 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
10681 ctcagaccct tttagtcagt gtggaaaatc tctagcaggg cccgtttaaa cccgctgatc
10741 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
10801 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
10861 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
10921 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
10981 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
11041 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
11101 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
11161 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
11221 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
11281 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
11341 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
11401 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
11461 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
11521 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
11581 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
11641 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
11701 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
11761 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
11821 ggatctgatc agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat
11881 acgacaaggt gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg
11941 cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact
12001 tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg
12061 tccaggacca ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg
12121 agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg
12181 ccatgaccga gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg
12241 gcaactgcgt gcacttcgtg gccgaggagc aggactgaca cgtgctacga gatttcgatt
12301 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
12361 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
12421 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
12481 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
12541 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
12601 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
12661 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
12721 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
12781 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
12841 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
12901 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
12961 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
13021 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
13081 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
13141 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
13201 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
13261 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
13321 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
13381 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
13441 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
13501 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
13561 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
13621 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
13681 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
13741 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
13801 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
13861 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
13921 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
13981 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
14041 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
14101 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
14161 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
14221 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
14281 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
14341 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
14401 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
14461 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
14521 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
14581 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
14641 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
14701 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
14761 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
14821 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
14881 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct
14941 tcgcgatgta cgggccagat atacgcgtt
//
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SuperFiCas9-tRNA-gRNA-all-in-One质粒载体应用举例
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