| 质粒名称: | pAct:dCas9-VPR |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 78898 |
| 存储实验室: | Norbert Perrimon |
|
载体骨架基本信息 |
|
| 载体名称: | pAWG |
| 载体抗性: | 氨苄霉素 |
| 载体类型: | 昆虫表达，CRISPR |
| 高拷贝/低拷贝: | 未知 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | dCas9-VPR |
| 插入大小: | 5733 bp |
| 启动子 | pActin (Drosophila) |
| 克隆方法: | Gibson Cloning |
| 5' 测序引物 | TTTGAACGCGACTTGAGAGC |
|
文献引用方法 |
|
| 材料和方法部分: | pAct:dCas9-VPR was a gift from Norbert Perrimon (爱迪基因 plasmid # 78898) |
| 参考文献部分: | Comparison of Cas9 activators in multiple species. Chavez A, Tuttle M, Pruitt BW, Ewen-Campen B, Chari R, Ter-Ovanesyan D, Haque SJ, Cecchi RJ, Kowal EJ, Buchthal J, Housden BE, Perrimon N, Collins JJ, Church G. Nat Methods. 2016 May 23. doi: 10.1038/nmeth.3871. 10.1038/nmeth.3871 PubMed 27214048 |
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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 pAct:dCas9-VPR 11968 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION Expresses dCas9-VPR under Actin promoter for CRISPRa in Drosophila
cell culture. Please note- this plasmid does not express GFP..
ACCESSION .
VERSION .
KEYWORDS pAct:dCas9-VPR
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11968)
AUTHORS Chavez A, Tuttle M, Pruitt BW, Ewen-Campen B, Chari R, Ter-Ovanesyan
D, Haque SJ, Cecchi RJ, Kowal EJ, Buchthal J, Housden BE, Perrimon
N, Collins JJ, Church G
TITLE Comparison of Cas9 activators in multiple species.
JOURNAL Nat Methods. 2016 May 23. doi: 10.1038/nmeth.3871.
PUBMED 27214048
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|pAct:dCas9-VPR|
FEATURES Location/Qualifiers
source 1..11968
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 140..2649
/label=Ac5 promoter
/note="Drosophila melanogaster actin 5C promoter"
protein_bind 2667..2691
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
regulatory 2697..2706
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2703..6806
/codon_start=1
/product="catalytically dead mutant of the Cas9
endonuclease from the Streptococcus pyogenes Type II
CRISPR/Cas system"
/label=Cas9m4
/note="RNA-guided DNA-binding protein that lacks
endonuclease activity due to the mutations D10A, D839A,
H840A, and N863A (Mali et al., 2013)"
/translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVAAIVPQSFLKDDSIDNKVLTRSDKARGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
CDS 6819..6839
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 6864..7013
/codon_start=1
/product="tetrameric repeat of the minimal activation
domain of herpes simplex virus VP16 (Beerli et al., 1998)"
/label=VP64
/translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
DLDML"
CDS 7038..7058
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7488..7844
/codon_start=1
/product="transcriptional activation domain of human RelA,
also known as p65 (O'Shea and Perkins, 2008)"
/label=RelA (p65) AD
/translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
EDFSSIADMDFSALL"
CDS 7863..8432
/codon_start=1
/product="transcriptional activation domain from the human
herpesvirus 4 (Epstein-Barr virus) replication and
transcription activator Rta/BRLF1 (Hardwick et al., 1992;
Chavez et al., 2015)"
/label=Rta AD
/translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
DTFLNDECLLHAMHISTGLSIFDTSLF"
protein_bind complement(8437..8461)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
regulatory 8469..8478
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 8475..9191
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(8520..8541)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(8781..8800)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 9128..9149
/label=EGFP-C
/note="EGFP, forward primer"
intron 9296..9361
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 9491..9511
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 9936..10070
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(9973..9992)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 10027..10046
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
CDS 10167..11027
/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(10385..10404)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 11198..11786
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 11687..11706
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 11940..11957
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcaggcaact
61 cgtgaaaggt aggcggatca gcggccgcag catgcaattc tatattctaa aaacacaaat
121 gatacttcta aaaaaaaatc atgaatggca tcaactctga atcaaatctt tgcagatgca
181 cctacttctc atttccactg tcacatcatt tttccagatc tcgctgcctg ttatgtggcc
241 cacaaaccaa gacacgtttt atggccatta aagctggctg atcgtcgcca aacaccaaat
301 acataatgaa tatgtacaca ttcgagaaag aagcgatcaa agaagcgtct tcgggcggag
361 taggagaatg cggaggagaa ggagaacgag ctgatctagt atctctccac aatccaatgc
421 caactgacca actggccata ttcggagcaa tttgaagcca atttccatcg cctggcgatc
481 gctccattct tggctatatg tttttcaccg ttacccgggg ccattttcaa agactcgtcg
541 gcaagataag attgtgtcac tcgctgtctc tcttcatttg tcgaagaatg ctgaggaatt
601 tcgcgatgac gtcggcgagt attttgaaga atgagaataa tttgtattta tacgaaaatc
661 agttagtgga attttctaca aaaacatgtt atctatagat aattttgttg caaaatatgt
721 tgactatgac aaagattgta tgtatatacc tttaatgtat tctcattttc ttatgtattt
781 ataatggcaa tgatgatact gatgatattt taagatgatg ccagaccaaa aggcttgaat
841 ttctgcgtct tttgccgaac gcagtgcatg tgcaattgtt gttttttgga atattcaatt
901 ttcggactgt ccgctttgat ttcagtttct tggcttattc aaaaagcaaa gtaaagccaa
961 aaaagcgaga tggcaatacc aaatgcggca aaacggtagt ggaaggaaag gggtgcgggg
1021 cagcggaagg aagggtgggg cggggcgtgg cggggtctgt ggctgggcgc gacgtcaccg
1081 acgttggagc cactcctttg accatgtgtg cgtgtgtgta ttattcgtgt ctcgccactc
1141 gccggttgtt tttttctttt tatgctgcgc tctctctagc gccatctcgc ttacgcatgc
1201 tcaacgcacc gcatgttgcc gtttcctttt atgcgtcatt ttggctcgaa ataggcaatt
1261 atttaaacaa agattagtca acgaaaacgc taaaataaat aagtctacaa tatggttact
1321 tattgccatg tgtgtgcagc caacgatagc aacaaaagca acaacacagg tggctttccc
1381 tctttcactt tttgtttgca agccgcgtgc gagcaagacg gcacgaccgg caaacgcaat
1441 tacgctgaca aagagcagac gaagttttgg cgaaaaacat caaggcgcct gatacgaatg
1501 catttgcaat aacaattgcg atatttaata ttgtttatga agctgtttga cttcaaaaca
1561 cacaaaaaaa aaaataaaac aaattatttg aaagagaatt aggaatcgga cgcttatcgt
1621 tagggtaaca acaagaaatg cttactgagt cacagcctct ggaaaactgc cgcaagccag
1681 agagagagag aaaaagaggg agagcagctt agaccgcatg tgcttgtgtg tgaggcgtct
1741 ctctcttcgt ctctgttgcg caaacgcata gactgcactg agaaaatcga ttacctattt
1801 tttatgaatg aatatttgca ctattactat tcaaaactat taagatagca atcacattca
1861 atagccaaat actataccac ctgagcgatg caacgaaatg atcaatttga gcaaaaatgc
1921 tgcatattta ggacggcatc attatagaaa tgcttcttgc tgtgtacttt tctctcgtct
1981 ggcagctgtt tcgccgttat tgttaaaacc ggcttaagtt aggtgtgttt tctacgacta
2041 gtgaatgccc tactagaaga tgtgtgttgc acaaaatgtc cctggaataa ccaatttgaa
2101 gtgcagatag cagtaaacgt aagctaatat gaatattatt taactgtaat gttttaatat
2161 cgctggacat tactaataaa cccactataa acacatgtac atatgtatgt tttggcatac
2221 aatgagtagt tggggaaaaa atgtgtaaaa gcaccgtgac catcacagca taaagataac
2281 cagctgaagt atcgaatatg agtaaccccc aaattgaatc acatgccgca actgatagga
2341 cccatggaag tacactcttc atggcgatat acaagacaca cacaagcacg aacacccagt
2401 tgcggaggaa attctccgta aatgaaaacc caatcggcga acaattcata cccatatatg
2461 gtaaaagttt tgaacgcgac ttgagagcgg agagcattgc ggctgataag gttttagcgc
2521 taagcgggct ttataaaacg ggctgcggga ccagttttca tatcactacc gtttgagttc
2581 ttgtgctgtg tggatactcc tcccgacaca aagccgctcc atcagccagc agtcgtctaa
2641 tccagagacc ccggatccag atatcgacaa gtttgtacaa aaaagcaggc tacaaagcca
2701 ccatggacaa gaagtactcc attgggctcg ctatcggcac aaacagcgtc ggctgggccg
2761 tcattacgga cgagtacaag gtgccgagca aaaaattcaa agttctgggc aataccgatc
2821 gccacagcat aaagaagaac ctcattggcg ccctcctgtt cgactccggg gagacggccg
2881 aagccacgcg gctcaaaaga acagcacggc gcagatatac ccgcagaaag aatcggatct
2941 gctacctgca ggagatcttt agtaatgaga tggctaaggt ggatgactct ttcttccata
3001 ggctggagga gtcctttttg gtggaggagg ataaaaagca cgagcgccac ccaatctttg
3061 gcaatatcgt ggacgaggtg gcgtaccatg aaaagtaccc aaccatatat catctgagga
3121 agaagcttgt agacagtact gataaggctg acttgcggtt gatctatctc gcgctggcgc
3181 atatgatcaa atttcgggga cacttcctca tcgaggggga cctgaaccca gacaacagcg
3241 atgtcgataa actctttatc caactggttc agacttacaa tcagcttttc gaagagaacc
3301 cgatcaacgc atccggagtt gacgccaaag caatcctgag cgctaggctg tccaaatccc
3361 ggcggctcga aaacctcatc gcacagctcc ctggggagaa gaagaacggc ctgtttggta
3421 atcttatcgc cctgtcactc gggctgaccc ccaactttaa atctaacttc gacctggccg
3481 aagatgccaa gcttcaactg agcaaagaca cctacgatga tgatctcgac aatctgctgg
3541 cccagatcgg cgaccagtac gcagaccttt ttttggcggc aaagaacctg tcagacgcca
3601 ttctgctgag tgatattctg cgagtgaaca cggagatcac caaagctccg ctgagcgcta
3661 gtatgatcaa gcgctatgat gagcaccacc aagacttgac tttgctgaag gcccttgtca
3721 gacagcaact gcctgagaag tacaaggaaa ttttcttcga tcagtctaaa aatggctacg
3781 ccggatacat tgacggcgga gcaagccagg aggaatttta caaatttatt aagcccatct
3841 tggaaaaaat ggacggcacc gaggagctgc tggtaaagct taacagagaa gatctgttgc
3901 gcaaacagcg cactttcgac aatggaagca tcccccacca gattcacctg ggcgaactgc
3961 acgctatcct caggcggcaa gaggatttct accccttttt gaaagataac agggaaaaga
4021 ttgagaaaat cctcacattt cggataccct actatgtagg ccccctcgcc cggggaaatt
4081 ccagattcgc gtggatgact cgcaaatcag aagagaccat cactccctgg aacttcgagg
4141 aagtcgtgga taagggggcc tctgcccagt ccttcatcga aaggatgact aactttgata
4201 aaaatctgcc taacgaaaag gtgcttccta aacactctct gctgtacgag tacttcacag
4261 tttataacga gctcaccaag gtcaaatacg tcacagaagg gatgagaaag ccagcattcc
4321 tgtctggaga gcagaagaaa gctatcgtgg acctcctctt caagacgaac cggaaagtta
4381 ccgtgaaaca gctcaaagaa gactatttca aaaagattga atgtttcgac tctgttgaaa
4441 tcagcggagt ggaggatcgc ttcaacgcat ccctgggaac gtatcacgat ctcctgaaaa
4501 tcattaaaga caaggacttc ctggacaatg aggagaacga ggacattctt gaggacattg
4561 tcctcaccct tacgttgttt gaagataggg agatgattga agaacgcttg aaaacttacg
4621 ctcatctctt cgacgacaaa gtcatgaaac agctcaagag gcgccgatat acaggatggg
4681 ggcggctgtc aagaaaactg atcaatggga tccgagacaa gcagagtgga aagacaatcc
4741 tggattttct taagtccgat ggatttgcca accggaactt catgcagttg atccatgatg
4801 actctctcac ctttaaggag gacatccaga aagcacaagt ttctggccag ggggacagtc
4861 ttcacgagca catcgctaat cttgcaggta gcccagctat caaaaaggga atactgcaga
4921 ccgttaaggt cgtggatgaa ctcgtcaaag taatgggaag gcataagccc gagaatatcg
4981 ttatcgagat ggcccgagag aaccaaacta cccagaaggg acagaagaac agtagggaaa
5041 ggatgaagag gattgaagag ggtataaaag aactggggtc ccaaatcctt aaggaacacc
5101 cagttgaaaa cacccagctt cagaatgaga agctctacct gtactacctg cagaacggca
5161 gggacatgta cgtggatcag gaactggaca tcaatcggct ctccgactac gacgtggctg
5221 ctatcgtgcc ccagtctttt ctcaaagatg attctattga taataaagtg ttgacaagat
5281 ccgataaagc tagagggaag agtgataacg tcccctcaga agaagttgtc aagaaaatga
5341 aaaattattg gcggcagctg ctgaacgcca aactgatcac acaacggaag ttcgataatc
5401 tgactaaggc tgaacgaggt ggcctgtctg agttggataa agccggcttc atcaaaaggc
5461 agcttgttga gacacgccag atcaccaagc acgtggccca aattctcgat tcacgcatga
5521 acaccaagta cgatgaaaat gacaaactga ttcgagaggt gaaagttatt actctgaagt
5581 ctaagctggt ctcagatttc agaaaggact ttcagtttta taaggtgaga gagatcaaca
5641 attaccacca tgcgcatgat gcctacctga atgcagtggt aggcactgca cttatcaaaa
5701 aatatcccaa gcttgaatct gaatttgttt acggagacta taaagtgtac gatgttagga
5761 aaatgatcgc aaagtctgag caggaaatag gcaaggccac cgctaagtac ttcttttaca
5821 gcaatattat gaattttttc aagaccgaga ttacactggc caatggagag attcggaagc
5881 gaccacttat cgaaacaaac ggagaaacag gagaaatcgt gtgggacaag ggtagggatt
5941 tcgcgacagt ccggaaggtc ctgtccatgc cgcaggtgaa catcgttaaa aagaccgaag
6001 tacagaccgg aggcttctcc aaggaaagta tcctcccgaa aaggaacagc gacaagctga
6061 tcgcacgcaa aaaagattgg gaccccaaga aatacggcgg attcgattct cctacagtcg
6121 cttacagtgt actggttgtg gccaaagtgg agaaagggaa gtctaaaaaa ctcaaaagcg
6181 tcaaggaact gctgggcatc acaatcatgg agcgatcaag cttcgaaaaa aaccccatcg
6241 actttctcga ggcgaaagga tataaagagg tcaaaaaaga cctcatcatt aagcttccca
6301 agtactctct ctttgagctt gaaaacggcc ggaaacgaat gctcgctagt gcgggcgagc
6361 tgcagaaagg taacgagctg gcactgccct ctaaatacgt taatttcttg tatctggcca
6421 gccactatga aaagctcaaa gggtctcccg aagataatga gcagaagcag ctgttcgtgg
6481 aacaacacaa acactacctt gatgagatca tcgagcaaat aagcgaattc tccaaaagag
6541 tgatcctcgc cgacgctaac ctcgataagg tgctttctgc ttacaataag cacagggata
6601 agcccatcag ggagcaggca gaaaacatta tccacttgtt tactctgacc aacttgggcg
6661 cgcctgcagc cttcaagtac ttcgacacca ccatagacag aaagcggtac acctctacaa
6721 aggaggtcct ggacgccaca ctgattcatc agtcaattac ggggctctat gaaacaagaa
6781 tcgacctctc tcagctcggt ggagacagca gggctgaccc caagaagaag aggaaggtgg
6841 aggccagcgg ttccggacgg gctgacgcat tggacgattt tgatctggat atgctgggaa
6901 gtgacgccct cgatgatttt gaccttgaca tgcttggttc ggatgccctt gatgactttg
6961 acctcgacat gctcggcagt gacgcccttg atgatttcga cctggacatg ctgattaact
7021 ctagaagttc cggatctccg aaaaagaaac gcaaagttgg tagccagtac ctgcccgaca
7081 ccgacgaccg gcaccggatc gaggaaaagc ggaagcggac ctacgagaca ttcaagagca
7141 tcatgaagaa gtcccccttc agcggcccca ccgaccctag acctccacct agaagaatcg
7201 ccgtgcccag cagatccagc gccagcgtgc caaaacctgc cccccagcct taccccttca
7261 ccagcagcct gagcaccatc aactacgacg agttccctac catggtgttc cccagcggcc
7321 agatctctca ggcctctgct ctggctccag cccctcctca ggtgctgcct caggctcctg
7381 ctcctgcacc agctccagcc atggtgtctg cactggctca ggcaccagca cccgtgcctg
7441 tgctggctcc tggacctcca caggctgtgg ctccaccagc ccctaaacct acacaggccg
7501 gcgagggcac actgtctgaa gctctgctgc agctgcagtt cgacgacgag gatctgggag
7561 ccctgctggg aaacagcacc gatcctgccg tgttcaccga cctggccagc gtggacaaca
7621 gcgagttcca gcagctgctg aaccagggca tccctgtggc ccctcacacc accgagccca
7681 tgctgatgga ataccccgag gccatcaccc ggctcgtgac aggcgctcag aggcctcctg
7741 atccagctcc tgcccctctg ggagcaccag gcctgcctaa tggactgctg tctggcgacg
7801 aggacttcag ctctatcgcc gatatggatt tctcagcctt gctgggctct ggcagcggca
7861 gccgggattc cagggaaggg atgtttttgc cgaagcctga ggccggctcc gctattagtg
7921 acgtgtttga gggccgcgag gtgtgccagc caaaacgaat ccggccattt catcctccag
7981 gaagtccatg ggccaaccgc ccactccccg ccagcctcgc accaacacca accggtccag
8041 tacatgagcc agtcgggtca ctgaccccgg caccagtccc tcagccactg gatccagcgc
8101 ccgcagtgac tcccgaggcc agtcacctgt tggaggatcc cgatgaagag acgagccagg
8161 ctgtcaaagc ccttcgggag atggccgata ctgtgattcc ccagaaggaa gaggctgcaa
8221 tctgtggcca aatggacctt tcccatccgc ccccaagggg ccatctggat gagctgacaa
8281 ccacacttga gtccatgacc gaggatctga acctggactc acccctgacc ccggaattga
8341 acgagattct ggataccttc ctgaacgacg agtgcctctt gcatgccatg catatcagca
8401 caggactgtc catcttcgac acatctctgt tttgaaaccc agctttcttg tacaaagtgg
8461 tgagctccgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
8521 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
8581 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
8641 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
8701 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
8761 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
8821 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
8881 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
8941 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
9001 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
9061 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
9121 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
9181 agctgtacaa gcaccggtcc acgtgacgta agctagcagg atctttgtga aggaacctta
9241 cttctgtggt gtgacataat tggacaaact acctacagag atttaaagct ctaaggtaaa
9301 tataaaattt ttaagtgtat aatgtgttaa actactgatt ctaattgttt gtgtatttta
9361 gattccaacc tatggaactg atgaatggga gcagtggtgg aatgccttta atgaggaaaa
9421 cctgttttgc tcagaagaaa tgccatctag tgatgatgag gctactgctg actctcaaca
9481 ttctactcct ccaaaaaaga agagaaaggt agaagacccc aaggactttc cttcagaatt
9541 gctaagtttt ttgagtcatg ctgtgtttag taatagaact cttgcttgct ttgctattta
9601 caccacaaag gaaaaagctg cactgctata caagaaaatt atggaaaaat attctgtaac
9661 ctttataagt aggcataaca gttataatca taacatactg ttttttctta ctccacacag
9721 gcatagagtg tctgctatta ataactatgc tcaaaaattg tgtaccttta gctttttaat
9781 ttgtaaaggg gttaataagg aatatttgat gtatagtgcc ttgactagag atcataatca
9841 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
9901 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
9961 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
10021 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc ccgtttaaac
10081 tacgcgtaat tcaaacaggg ttctggcgtc gttctcgtac tgttttcccc aggccagtgc
10141 tttagcgtta ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
10201 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
10261 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
10321 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
10381 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
10441 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
10501 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
10561 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
10621 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
10681 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
10741 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
10801 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
10861 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
10921 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
10981 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
11041 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
11101 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
11161 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
11221 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
11281 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
11341 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
11401 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
11461 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
11521 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
11581 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
11641 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
11701 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
11761 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
11821 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
11881 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
11941 gcgagtcagt gagcgaggaa gcggaaga
//
