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- 详细信息
- 文献和实验
- 技术资料
- 保存条件：
-20℃
- 保质期：
6个月
- 英文名：
pCas-Guide-EF1a-GFP
- 库存：
大量
- 供应商：
钰博生物
- 规格：
1ug/5ug
pCas-Guide-EF1a-GFP
载体基本信息
| 出品公司: | Ybscience |
|---|---|
| 载体名称: | pCas-Guide-EF1a-GFP |
| 质粒类型: | CRISPR/Cas9载体；基因敲除载体；哺乳动物细胞载体 |
| 高拷贝/低拷贝: | 高拷贝 |
| 插入位点: | 限制性内切酶，多克隆位点 |
| 启动子: | U6 |
| 载体大小: | 10493 bp |
| 5' 测序引物及序列: | 5’ gatcgATGGGAGGTGGTATGGGAGGg 3’ |
| 3' 测序引物及序列: | 5’ aaaacCCTCCCATACCACCTCCCATc 3’ |
| 载体标签: | Myc(C-ter） |
| 载体抗性: | 氨苄青霉素 |
| 筛选标记: | GFP |
| 克隆菌株: | Stbl3 |
| 宿主细胞（系）: | 哺乳动物细胞 |
| 备注: | -- |
| 稳定性: | 瞬表达或稳表达 |
| 组成型/诱导型: | 组成型 |
| 病毒/非病毒: | 非病毒 |
载体质粒图谱和多克隆位点信息
载体简介
pCas-Guide-EF1a-GFP All-in-one vector Expresses human codon-optimized Cas9, C-terminal Myc-DDK tagged BamH I and BsmB I can be used for genomic target sequence cloning Expresses gRNA containing the inserted target sequence and the gRNA scaffold GFP as a reporter for tracking or sorting transfected cells CF3 primer can be used to sequence the target sequence after cloning
载体序列
LOCUS Exported File 10493 bp ds-DNA circular SYN 23-七月-2016
DEFINITION Mammalian vector for co-expressing a guide RNA (gRNA) together with
Cas9 and TurboGFP, for genome editing using the S. pyogenes
CRISPR/Cas9 system.
ACCESSION .
VERSION .
KEYWORDS pCas-Guide-EF1a-GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10493)
AUTHORS OriGene
TITLE Direct Submission
JOURNAL Exported 2016-07-23 from SnapGene Viewer 1.5.3
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..10493
/organism="synthetic DNA construct"
/lab_host="Mammalian Cells"
/mol_type="other DNA"
promoter 90..330
/note="U6"
/note="RNA polymerase III promoter for human U6 snRNA"
/note="color: #000000; direction: RIGHT"
misc_RNA 384..459
/note="gRNA scaffold"
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
/note="color: #00ccff; direction: RIGHT"
terminator 460..465
/note="polIII terminator"
/note="RNA polymerase III transcription terminator"
/note="color: #ffffff"
promoter 480..1658
/note="EF-1-alpha promoter"
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
/note="color: #000000; direction: RIGHT"
intron 711..1649
/note="EF-1-alpha intron A"
/note="intron upstream of the start codon of human
EF-1-alpha"
/note="color: #------"
CDS 1737..2432
/codon_start=1
/product="improved green fluorescent protein from
Pontellina plumata?(Evdokimov et al., 2006)"
/note="TurboGFP"
/note="mammalian codon-optimized"
/note="color: #05fd14"
/translation="MESDESGLPAMEIECRITGTLNGVEFELVGGGEGTPEQGRMTNKM
KSTKGALTFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVL
HVSFSYRYEAGRVIGDFKVMGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNDLDGSFTR
TFSLRDGGYYSSVVDSHMHFKSAIHPSILQNGGPMFAFRRVEEDHSNTELGIVEYQHAF
KTPDADAGEE"
enhancer 3047..3350
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
/note="color: #000000"
promoter 3351..3554
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
/note="color: #000000; direction: RIGHT"
promoter 3580..3598
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
/note="color: #ffffff; direction: RIGHT"
CDS 3657..7760
/codon_start=1
/locus_tag="
"
/product="Cas9 (Csn1) endonuclease from the?Streptococcus
pyogenes?Type II CRISPR/Cas system"
/note="Cas9"
/note="generates RNA-guided double strand breaks in DNA"
/note="color: #993366"
/protein_id="
"
/translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
CDS 7761..7781
/codon_start=1
/locus_tag="
"
/product="nuclear localization signal of SV40 large T
antigen"
/note="SV40 NLS"
/note="
"
/note="color: #993366"
/protein_id="
"
/translation="PKKKRKV"
CDS 7788..7808
/codon_start=1
/locus_tag="
"
/product="nuclear localization signal of SV40 large T
antigen"
/note="SV40 NLS"
/note="
"
/note="color: #993366"
/protein_id="
"
/translation="PKKKRKV"
CDS 7830..7859
/codon_start=1
/product="Myc (human c-Myc oncogene) epitope tag"
/note="Myc"
/note="color: #cc99b2"
/translation="EQKLISEEDL"
CDS 7878..7901
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/note="FLAG"
/note="color: #cc99b2"
/translation="DYKDDDDK"
polyA_signal 7948..8570
/note="hGH poly(A) signal"
/note="human growth hormone polyadenylation signal"
/note="color: #a6acb3"
rep_origin complement(8691..9279)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
/note="color: #ffff00"
CDS complement(9454..10314)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/note="This feature has 2 segments:
???1:??9454?..?10245?/?#cc99ff
???2:?10246?..?10314?/?#cc99ff?/?signal?sequence
Cleavage site after base 10245"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
ORIGIN
1 gaattcccca gtggaaagac gcgcaggcaa aacgcaccac gtgacggagc gtgaccgcgc
61 gccgagcgcg cgccaaggtc gggcaggaag agggcctatt tcccatgatt ccttcatatt
121 tgcatatacg atacaaggct gttagagaga taattagaat taatttgact gtaaacacaa
181 agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt
241 taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt
301 tcttgggttt atatatcttg tggaaaggac gcgggatcca ctggaccagg cagcagcgtc
361 agaagacttt tttggaacgt ctcgttttag agctagaaat agcaagttaa aataaggcta
421 gtccgttatc aacttgaaaa agtggcaccg agtcggtgct ttttttggtg tacacgtgag
481 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
541 gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
601 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
661 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg
721 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta
781 cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg
841 agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc
901 ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt
961 tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc
1021 aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg
1081 cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag
1141 cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg
1201 gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag
1261 ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga
1321 cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt
1381 cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt
1441 agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg
1501 agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat
1561 tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag
1621 tggttcaaag tttttttctt ccatttcagg tgtcgtgact atagggcggc cggacgtgac
1681 aaatggaagt agcacgcctc actaggctcg tgcagatgga cagcaccgct gcagccatgg
1741 agagcgacga gagcggcctg cccgccatgg agatcgagtg ccgcatcacc ggcaccctga
1801 acggcgtgga gttcgagctg gtgggcggcg gagagggcac ccccgagcag ggccgcatga
1861 ccaacaagat gaagagcacc aaaggcgccc tgaccttcag cccctacctg ctgagccacg
1921 tgatgggcta cggcttctac cacttcggca cctaccccag cggctacgag aaccccttcc
1981 tgcacgccat caacaacggc ggctacacca acacccgcat cgagaagtac gaggacggcg
2041 gcgtgctgca cgtgagcttc agctaccgct acgaggccgg ccgcgtgatc ggcgacttca
2101 aggtgatggg caccggcttc cccgaggaca gcgtgatctt caccgacaag atcatccgca
2161 gcaacgccac cgtggagcac ctgcacccca tgggcgataa cgatctggat ggcagcttca
2221 cccgcacctt cagcctgcgc gacggcggct actacagctc cgtggtggac agccacatgc
2281 acttcaagag cgccatccac cccagcatcc tgcagaacgg gggccccatg ttcgccttcc
2341 gccgcgtgga ggaggatcac agcaacaccg agctgggcat cgtggagtac cagcacgcct
2401 tcaagacccc ggatgcagat gccggtgaag aaagagttta atcgatgata tcagatcccc
2461 gggatgcaga aattgatgat ctattaaaca ataaagatgt ccactaaaat ggaagttttt
2521 cctgtcatac tttgttaaga agggtgagaa cagagtacct acattttgaa tggaaggatt
2581 ggagctacgg gggtgggggt ggggtgggat tagataaatg cctgctcttt actgaaggct
2641 ctttactatt gctttatgat aatgtttcat agttggatat cataatttaa acaagcaaaa
2701 ccaaattaag ggccagctca ttcctcccac tcatgatcta tagatctata gatctctcgt
2761 gggatcattg tttttctctt gattcccact ttgtggttct aagtactgtg gtttccaaat
2821 gtgtcagttt catagcctga agaacgagat cagcagcctc tgttccacat acacttcatt
2881 ctcagtattg ttttgccaag ttctaattcc atcagaagct ggtcgagatc cggaaccctt
2941 aatataactt cgtataatgt atgctatacg aagttattag gtccactagt tattaatagt
3001 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
3061 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
3121 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
3181 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt ccgcccccta
3241 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttacggg
3301 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
3361 tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
3421 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
3481 gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
3541 atataagcag agctcgttta gtgaaccgtc agaattttgt aatacgactc actatagggc
3601 ggccgggaat tcgtcgactg gaaccggtac cgaggagatc tgccgccgcg atcgccatgg
3661 ataagaaata ctcaatagga ctggatattg gcacaaatag cgtcggatgg gctgtgatca
3721 ctgatgaata taaggttcct tctaaaaagt tcaaggttct gggaaataca gaccgccaca
3781 gtatcaaaaa aaatcttata ggggctcttc tgtttgacag tggagagaca gccgaagcta
3841 ctagactcaa acggacagct aggagaaggt atacaagacg gaagaatagg atttgttatc
3901 tccaggagat tttttcaaat gagatggcca aagtggatga tagtttcttt catagacttg
3961 aagagtcttt tttggtggaa gaagacaaga agcatgaaag acatcctatt tttggaaata
4021 tagtggatga agttgcttat cacgagaaat atccaactat ctatcatctg agaaaaaaat
4081 tggtggattc tactgataaa gccgatttgc gcctgatcta tttggccctg gcccacatga
4141 ttaagtttag aggtcatttt ttgattgagg gcgatctgaa tcctgataat agtgatgtgg
4201 acaaactgtt tatccagttg gtgcaaacct acaatcaact gtttgaagaa aaccctatta
4261 acgcaagtgg agtggatgct aaagccattc tttctgcaag attgagtaaa tcaagaagac
4321 tggaaaatct cattgctcag ctccccggtg agaagaaaaa tggcctgttt gggaatctca
4381 ttgctttgtc attgggtttg acccctaatt ttaaatcaaa ttttgatttg gcagaagatg
4441 ctaaactcca gctttcaaaa gatacttacg atgatgatct ggataatctg ttggctcaaa
4501 ttggagatca atatgctgat ttgtttttgg cagctaagaa tctgtcagat gctattctgc
4561 tttcagacat cctgagagtg aatactgaaa taactaaggc tcccctgtca gcttcaatga
4621 ttaaacgcta cgatgaacat catcaagact tgactcttct gaaagccctg gttagacaac
4681 aacttccaga aaagtataaa gaaatctttt ttgatcaatc aaaaaacgga tatgcaggtt
4741 atattgatgg cggcgcaagc caagaagaat tttataaatt tatcaaacca attctggaaa
4801 aaatggatgg tactgaggaa ctgttggtga aactgaatag agaagatttg ctgcgcaagc
4861 aacggacctt tgacaacggc tctattcccc atcaaattca cttgggtgag ctgcatgcta
4921 ttttgagaag acaagaagac ttttatccat ttctgaaaga caatagagag aagattgaaa
4981 aaatcttgac ttttaggatt ccttattatg ttggtccatt ggccagaggc aatagtaggt
5041 ttgcatggat gactcggaag tctgaagaaa caattacccc atggaatttt gaagaagttg
5101 tcgataaagg tgcttcagct caatcattta ttgaacgcat gacaaacttt gataaaaatc
5161 ttccaaatga aaaagtgctg ccaaaacata gtttgcttta tgagtatttt accgtttata
5221 acgaattgac aaaggtcaaa tatgttactg aaggaatgag aaaaccagca tttctttcag
5281 gtgaacagaa gaaagccatt gttgatctgc tcttcaaaac aaataggaaa gtgaccgtta
5341 agcaactgaa agaagattat ttcaaaaaaa tagaatgttt tgatagtgtt gaaatttcag
5401 gagttgaaga tagatttaat gcttcactgg gtacatacca tgatttgctg aaaattatta
5461 aagataaaga ttttttggat aatgaagaaa atgaagacat cctggaggat attgttctga
5521 cattgaccct gtttgaagat agggagatga ttgaggaaag acttaaaaca tacgctcacc
5581 tctttgatga taaggtgatg aaacagctta aaagacgcag atatactggt tggggaaggt
5641 tgtccagaaa attgattaat ggtattaggg ataagcaatc tggcaaaaca atactggatt
5701 ttttgaaatc agatggtttt gccaatcgca attttatgca gctcatccat gatgatagtt
5761 tgacatttaa agaagacatc caaaaagcac aagtgtctgg acaaggcgat agtctgcatg
5821 aacatattgc aaatctggct ggtagccctg ctattaaaaa aggtattctc cagactgtga
5881 aagttgttga tgaattggtc aaagtgatgg ggcggcataa gccagaaaat atcgttattg
5941 aaatggcaag agaaaatcag acaactcaaa agggccagaa aaattccaga gagaggatga
6001 aaagaatcga agaaggtatc aaagaactgg gaagtcagat tcttaaagag catcctgttg
6061 aaaatactca attgcaaaat gaaaagctct atctctatta tctccaaaat ggaagagata
6121 tgtatgtgga ccaagaactg gatattaata ggctgagtga ttatgatgtc gatcacattg
6181 ttccacaaag tttccttaaa gacgattcaa tagacaataa ggtcctgacc aggtctgata
6241 aaaatagagg taaatccgat aacgttccaa gtgaagaagt ggtcaaaaag atgaaaaact
6301 attggagaca acttctgaac gccaagctga tcactcaaag gaagtttgat aatctgacca
6361 aagctgaaag aggaggtttg agtgaacttg ataaagctgg ttttatcaaa cgccaattgg
6421 ttgaaactcg ccaaatcact aagcatgtgg cacaaatttt ggatagtcgc atgaatacta
6481 aatacgatga aaatgataaa cttattagag aggttaaagt gattaccctg aaatctaaac
6541 tggtttctga cttcagaaaa gatttccaat tctataaagt gagagagatt aacaattacc
6601 atcatgccca tgatgcctat ctgaatgccg tcgttggaac tgctttgatt aagaaatatc
6661 caaaacttga aagcgagttt gtctatggtg attataaagt ttatgatgtt aggaaaatga
6721 ttgctaagtc tgagcaagaa ataggcaaag caaccgcaaa gtatttcttt tactctaata
6781 tcatgaactt cttcaaaaca gaaattacac ttgcaaatgg agagattcgc aaacgccctc
6841 tgatcgaaac taatggggaa actggagaaa ttgtctggga taaagggaga gattttgcca
6901 cagtgcgcaa agtgttgtcc atgccccaag tcaatatcgt caagaaaaca gaagtgcaga
6961 caggcggatt ctctaaggag tcaattctgc caaaaagaaa ttccgacaag ctgattgcta
7021 ggaaaaaaga ctgggaccca aaaaaatatg gtggttttga tagtccaacc gtggcttatt
7081 cagtcctggt ggttgctaag gtggaaaaag ggaaatccaa gaagctgaaa tccgttaaag
7141 agctgctggg gatcacaatt atggaaagaa gttcctttga aaaaaatccc attgactttc
7201 tggaagctaa aggatataag gaagttaaaa aagacctgat cattaaactg cctaaatata
7261 gtctttttga gctggaaaac ggtaggaaac ggatgctggc tagtgccgga gaactgcaaa
7321 aaggaaatga gctggctctg ccaagcaaat atgtgaattt tctgtatctg gctagtcatt
7381 atgaaaagtt gaagggtagt ccagaagata acgaacaaaa acaattgttt gtggagcagc
7441 ataagcatta tctggatgag attattgagc aaatcagtga attttctaag agagttattc
7501 tggcagatgc caatctggat aaagttctta gtgcatataa caaacataga gacaaaccaa
7561 taagagaaca agcagaaaat atcattcatc tgtttacctt gaccaatctt ggagcacccg
7621 ctgcttttaa atactttgat acaacaattg ataggaaaag atatacctct acaaaagaag
7681 ttctggatgc cactcttatc catcaatcca tcactggtct ttatgaaaca cgcattgatt
7741 tgagtcagct gggaggtgac cccaagaaaa aacgcaaggt ggaagatcct aagaaaaagc
7801 ggaaagtgga cacgcgtacg cggccgctcg agcagaaact catctcagaa gaggatctgg
7861 cagcaaatga tatcctggat tacaaggatg acgacgataa ggtttaaacg gccggccgcg
7921 gtcatagctg tttcctgaac agatcccggg tggcatccct gtgacccctc cccagtgcct
7981 ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt
8041 tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta
8101 tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca
8161 agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg
8221 attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc
8281 taatttttgt ttttttggta gaggcggggt ttcaccatat tggccaggct ggtctccaac
8341 tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga
8401 accactgctc ccttccctgt ccttctgatt ttaaaataac tataccagca ggaggacgtc
8461 cagacacagc ataggctacc tggccatgcc caaccggtgg gacatttgag ttgcttgctt
8521 ggcactgtcc tctcatgcgt tgggtccact cagtagatgc ctgttgaatt gggtacgcgg
8581 ccagcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
8641 ggataacgca ggaaagaaca tgtccgtaaa aaggccgcgt tgctggcgtt tttccatagg
8701 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
8761 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
8821 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
8881 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
8941 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
9001 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
9061 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
9121 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
9181 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
9241 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
9301 acggggtctg acgctcagtg gaacgacgcg taactcacgt taagggattt tggtcatgag
9361 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
9421 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
9481 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
9541 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
9601 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
9661 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
9721 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
9781 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
9841 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
9901 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
9961 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
10021 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
10081 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
10141 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
10201 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
10261 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
10321 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgatga tattatttta
10381 tcttgtgcaa tgtaacatca gagattttga gacacgggcc agagctgcca ggaaacagct
10441 atgaccatgt aatacgactc actatagggg atatcagctg gatggcagtt aac
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pCas-Guide-EF1a-GFP
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