| 质粒名称: | pCAG-eCas9-GFP-U6-gRNA |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 79145 |
| 存储实验室: | Jizhong Zou |
|
载体骨架基本信息 |
|
| 载体名称: | eSpCas9(1.1) |
| 载体出品公司: | Addgene #71814 |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 10243 bp |
| 载体修饰: | 人细胞色素c氧化酶亚基VIII（human cytochrome C oxidase subunit VIII） |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 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: | eSpCas9(1.1) |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pCAG-eCas9-GFP-U6-gRNA was a gift from Jizhong Zou (Addgene plasmid # 79145 ; http://n2t.net/addgene:79145 ; RRID:Addgene_79145) |
| 参考文献部分: | -- |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pCAG-eCas9-GFP-U6-gRNA 10243 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION All-in-one CRISPR/Cas9 vector with high-fidelity eSpCas9 expression
in human pluripotent stem cells.
ACCESSION .
VERSION .
KEYWORDS pCAG-eCas9-GFP-U6-gRNA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10243)
TITLE pCAG-SpCas9-U6-gRNA
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 10243)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..10243
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..241
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 1..21
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 172..191
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 268..343
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
enhancer 523..826
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 828..1105
/label=chicken beta-actin promoter
intron 1107..2124
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 2048..2070
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 2132..2151
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
regulatory 2199..2208
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2208..2273
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 2280..2300
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 2325..6425
/codon_start=1
/product="Cas9 endonuclease from the Streptococcus pyogenes
Type II CRISPR/Cas system, mutated to improve targeting
specificity (Slaymaker et al., 2016)"
/label=eSpCas9(1.1)
/note="carries the mutations K848A, K1003A, and R1060A"
/translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLADDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPALESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKAPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 6426..6473
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 6489..6542
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS 6543..7256
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
primer_bind complement(6585..6606)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(6846..6865)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 7193..7214
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(7284..7301)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 7290..7497
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 7506..7646
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 7506..7635
/label=AAV2 ITR
rep_origin 7721..8176
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7808..7827)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 8018..8039
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(8193..8212)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 8312..8334
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(8372..8390)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 8458..8562
/gene="bla"
/label=AmpR promoter
CDS 8563..9423
/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 complement(8781..8800)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 9594..10182
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 10083..10102
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
241 cgaaacaccg ggtcttcgag aagacctgtt ttagagctag aaatagcaag ttaaaataag
301 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg ttttagagct
361 agaaatagca agttaaaata aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac
421 aaatggctct agtaacggcc gccagtgtgc tggaattcag gctagttatt aatagtaatc
481 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
541 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
601 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
661 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
721 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
781 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc
841 cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt
901 tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggggc gcgcgccagg
961 cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa
1021 tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc ggcggcccta
1081 taaaaagcga agcgcgcggc gggcggggag tcgctgcgac gctgccttcg ccccgtgccc
1141 cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag
1201 gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg
1261 cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc
1321 ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc
1381 tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc
1441 agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag
1501 gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg
1561 tcggtcgggc tgcaaccccc cctgcacccc cctccccgag ttgctgagca cggcccggct
1621 tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg
1681 cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg ccggggaggg ctcgggggag
1741 gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg cggcgagccg cagccattgc
1801 cttttatggt aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag
1861 ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc
1921 gccggcagga aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt
1981 ctccctctcc agcctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc
2041 agggcggggt tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt
2101 catgccttct tctttttcct acagctcctg ggcaacgtgc tggttattgt gctgtctcat
2161 cattttggca aagaattcct gaattctgca gatccggtgc caccatggac tataaggacc
2221 acgacggaga ctacaaggat catgatattg attacaaaga cgatgacgat aagatggccc
2281 caaagaagaa gcggaaggtc ggtatccacg gagtcccagc agccgacaag aagtacagca
2341 tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg
2401 tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc
2461 tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa
2521 ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca
2581 gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg
2641 tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg
2701 cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg
2761 acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc
2821 acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc
2881 agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg
2941 acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg
3001 cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg
3061 gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga
3121 gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg
3181 ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga
3241 gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg
3301 agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt
3361 acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag
3421 ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg
3481 aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca
3541 acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg
3601 aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc
3661 gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca
3721 gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt
3781 ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg
3841 tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag
3901 tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg
3961 ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg
4021 actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt
4081 tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc
4141 tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg
4201 aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag
4261 tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga
4321 tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg
4381 gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg
4441 acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc
4501 tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc
4561 tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga
4621 accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg
4681 gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc
4741 agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg
4801 aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc
4861 tggccgacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga
4921 gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc
4981 tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg
5041 gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga
5101 tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg
5161 acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc
5221 ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg
5281 cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctgcg ctggaaagcg
5341 agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc
5401 aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg aactttttca
5461 agaccgagat taccctggcc aacggcgaga tccggaaggc gcctctgatc gagacaaacg
5521 gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc
5581 tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca
5641 aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg
5701 accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg
5761 ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca
5821 ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct
5881 acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg
5941 aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg
6001 ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg
6061 gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg
6121 acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc
6181 tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg
6241 agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact
6301 ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc
6361 tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag
6421 gcgacaaaag gccggcggcc acgaaaaagg ccggccaggc aaaaaagaaa aaggaattcg
6481 gcagtggaga gggcagagga agtctgctaa catgcggtga cgtcgaggag aatcctggcc
6541 cagtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
6601 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
6661 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
6721 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
6781 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
6841 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
6901 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
6961 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
7021 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
7081 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
7141 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
7201 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaaggaat
7261 tctaactaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
7321 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
7381 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
7441 ggtggggcag gacagcaagg gggaggattg ggaagagaat agcaggcatg ctggggagcg
7501 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
7561 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
7621 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
7681 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
7741 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
7801 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
7861 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
7921 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
7981 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
8041 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
8101 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
8161 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
8221 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
8281 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
8341 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
8401 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
8461 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
8521 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
8581 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
8641 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
8701 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
8761 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
8821 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
8881 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
8941 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
9001 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
9061 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
9121 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
9181 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
9241 tggtttattg ctgataaatc tggagccggt gagcgtggaa gccgcggtat cattgcagca
9301 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
9361 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
9421 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
9481 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
9541 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
9601 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
9661 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
9721 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
9781 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
9841 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
9901 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
9961 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
10021 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
10081 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
10141 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
10201 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
