| 质粒名称: | pDuRCC-N |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 81194 |
| 存储实验室: | Eckhard Boles |
|
载体骨架基本信息 |
|
| 载体名称: | pRS42N |
| 载体出品公司: | Euroscarf |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 酵母表达,CRISPR |
| 筛选标记: | Nourseothricin |
| 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: | Cas9 |
| 物种来源: | S. pyogenes |
| 插入大小: | -- |
| 基因突变: | WT - codon optimized |
| GenBank编号 | -- |
| 启动子 | ROX3 |
| 标签/融合蛋白: | NTS（主干上的C终端） |
| 克隆方法: | Gibson 克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pDuRCC-N was a gift from Eckhard Boles (Addgene plasmid # 81194 ; http://n2t.net/addgene:81194 ; RRID:Addgene_81194) |
| 参考文献部分: | Simplified CRISPR-Cas genome editing for Saccharomyces cerevisiae. Generoso WC, Gottardi M, Oreb M, Boles E. J Microbiol Methods. 2016 Aug;127:203-5. doi: 10.1016/j.mimet.2016.06.020. Epub 2016 Jun 17. 10.1016/j.mimet.2016.06.020 PubMed 27327211 |
买家导航
pDuRCCN载体基本信息
pDuRCCN载体图谱质粒图谱和多克隆位点信息
pDuRCCN载体简介
pDuRCCN载体序列
LOCUS pDuRCC-N 10325 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expression of Cas9 and two gRNA cassettes in S. cerevisiae;
Nourseothricin Resistance.
ACCESSION .
VERSION .
KEYWORDS pDuRCC-N
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10325)
AUTHORS Generoso WC, Gottardi M, Oreb M, Boles E
TITLE Simplified CRISPR-Cas genome editing for Saccharomyces cerevisiae.
JOURNAL J Microbiol Methods. 2016 Aug;127:203-5. doi:
10.1016/j.mimet.2016.06.020. Epub 2016 Jun 17.
PUBMED 27327211
REFERENCE 2 (bases 1 to 10325)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..10325
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(10..29)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 53..321
/label=SNR52 promoter
/note="promoter for the S. cerevisiae small nucleolar RNA
gene SNR52"
misc_RNA 345..419
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
terminator 424..443
/label=SUP4 terminator
/note="transcription terminator for the S. cerevisiae SUP4
tRNA gene"
primer_bind complement(496..514)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
rep_origin 555..1897
/label=2u ori
/note="yeast 2u plasmid origin of replication"
protein_bind 887..934
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
promoter 1924..2028
/gene="bla"
/label=AmpR promoter
CDS 2029..2889
/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(2247..2266)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 3060..3648
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3549..3568
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 3685..3953
/label=SNR52 promoter
/note="promoter for the S. cerevisiae small nucleolar RNA
gene SNR52"
misc_RNA 3977..4051
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
terminator 4056..4075
/label=SUP4 terminator
/note="transcription terminator for the S. cerevisiae SUP4
tRNA gene"
terminator 4096..4343
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
primer_bind 4309..4327
/label=CYC1
/note="CYC1 transcription termination signal, reverse
primer"
CDS complement(4359..4379)
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS complement(4392..8495)
/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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
gene complement(9184..10303)
/label=natMX6
/note="yeast selectable marker conferring nourseothricin
resistance"
terminator 9184..9381
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
CDS complement(9387..9959)
/codon_start=1
/gene="Streptomyces noursei nat1"
/product="nourseothricin acetyltransferase"
/label=NrsR
/note="confers resistance to nourseothricin"
/translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
LCGLDTALYDGTASDGEQALYMSMPCP"
promoter complement(9960..10303)
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
ORIGIN
1 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccgggta attctttgaa
61 aagataatgt atgattatgc tttcactcat atttatacag aaacttgatg ttttctttcg
121 agtatataca aggtgattac atgtacgttt gaagtacaac tctagatttt gtagtgccct
181 cttgggctag cggtaaaggt gcgcattttt tcacacccta caatgttctg ttcaaaagat
241 tttggtcaaa cgctgtagaa gtgaaagttg gtgcgcatgt ttcggcgttc gaaacttctc
301 cgcagtgaaa gataaatgat cggatccacg ttctctgcgg ccgcgtttta gagctagaaa
361 tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgg
421 tgcttttttt gttttttatg tctcggcgaa ttgggtgcgg ccgcgtttgt cacctttcat
481 aggcctgacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
541 tggtttctta gtatgatcca atatcaaagg aaatgatagc attgaaggat gagactaatc
601 caattgagga gtggcagcat atagaacagc taaagggtag tgctgaagga agcatacgat
661 accccgcatg gaatgggata atatcacagg aggtactaga ctacctttca tcctacataa
721 atagacgcat ataagtacgc atttaagcat aaacacgcac tatgccgttc ttctcatgta
781 tatatatata caggcaacac gcagatatag gtgcgacgtg aacagtgagc tgtatgtgcg
841 cagctcgcgt tgcattttcg gaagcgctcg ttttcggaaa cgctttgaag ttcctattcc
901 gaagttccta ttctctagaa agtataggaa cttcagagcg cttttgaaaa ccaaaagcgc
961 tctgaagacg cactttcaaa aaaccaaaaa cgcaccggac tgtaacgagc tactaaaata
1021 ttgcgaatac cgcttccaca aacattgctc aaaagtatct ctttgctata tatctctgtg
1081 ctatatccct atataaccta cccatccacc tttcgctcct tgaacttgca tctaaactcg
1141 acctctacat tttttatgtt tatctctagt attactcttt agacaaaaaa attgtagtaa
1201 gaactattca tagagtgaat cgaaaacaat acgaaaatgt aaacatttcc tatacgtagt
1261 atatagagac aaaatagaag aaaccgttca taattttctg accaatgaag aatcatcaac
1321 gctatcactt tctgttcaca aagtatgcgc aatccacatc ggtatagaat ataatcgggg
1381 atgcctttat cttgaaaaaa tgcacccgca gcttcgctag taatcagtaa acgcgggaag
1441 tggagtcagg ctttttttat ggaagagaaa atagacacca aagtagcctt cttctaacct
1501 taacggacct acagtgcaaa aagttatcaa gagactgcat tatagagcgc acaaaggaga
1561 aaaaaagtaa tctaagatgc tttgttagaa aaatagcgct ctcgggatgc atttttgtag
1621 aacaaaaaag aagtatagat tctttgttgg taaaatagcg ctctcgcgtt gcatttctgt
1681 tctgtaaaaa tgcagctcag attctttgtt tgaaaaatta gcgctctcgc gttgcatttt
1741 tgttttacaa aaatgaagca cagattcttc gttggtaaaa tagcgctttc gcgttgcatt
1801 tctgttctgt aaaaatgcag ctcagattct ttgtttgaaa aattagcgct ctcgcgttgc
1861 atttttgttc tacaaaatga agcacagatg cttcgttcag gtggcacttt tcggggaaat
1921 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
1981 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
2041 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
2101 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
2161 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
2221 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
2281 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
2341 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
2401 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
2461 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
2521 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
2581 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
2641 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
2701 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
2761 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
2821 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
2881 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
2941 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
3001 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
3061 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
3121 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
3181 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
3241 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
3301 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
3361 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
3421 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
3481 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
3541 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
3601 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
3661 gcgggctagg gataacaggg taattctttg aaaagataat gtatgattat gctttcactc
3721 atatttatac agaaacttga tgttttcttt cgagtatata caaggtgatt acatgtacgt
3781 ttgaagtaca actctagatt ttgtagtgcc ctcttgggct agcggtaaag gtgcgcattt
3841 tttcacaccc tacaatgttc tgttcaaaag attttggtca aacgctgtag aagtgaaagt
3901 tggtgcgcat gtttcggcgt tcgaaacttc tccgcagtga aagataaatg atcggatcca
3961 cgttctctgc ggccgcgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt
4021 atcaacttga aaaagtggca ccgagtcggt ggtgcttttt ttgtttttta tgtctcggcg
4081 aattgggtac cggccgcaaa ttaaagcctt cgagcgtccc aaaaccttct caagcaaggt
4141 tttcagtata atgttacatg cgtacacgcg tctgtacaga aaaaaaagaa aaatttgaaa
4201 tataaataac gttcttaata ctaacataac tataaaaaaa taaataggga cctagacttc
4261 aggttgtcta actccttcct tttcggttag agcggatgtg gggggagggc gtgaatgtaa
4321 gcgtgacata actaattaca tgactcgagg tcgacttaaa cctttctctt cttctttggg
4381 tcagctctag agtcaccacc caattgagac aagtcaattc tggtttcgta caaaccggta
4441 atagattggt gaatcaaggt agcgtccaaa acttccttgg tagaggtgta tctctttctg
4501 tcaatggtgg tgtcgaagta cttgaaagca gctggagcac ccaagttggt caaggtgaac
4561 aagtgaataa tgttttcagc ttgttctcta attggcttgt ctctgtgctt gttgtaagca
4621 gacaaaacct tgtccaagtt agcgtcagcc aaaataactc tcttagagaa ttcagaaatt
4681 tgttcaataa tttcgtccaa gtagtgcttg tgttgttcaa cgaacaattg cttttgttcg
4741 ttgtcttctg gagaaccctt caacttttcg tagtgagaag ccaagtacaa gaagttaacg
4801 tacttagatg gcaaagccaa ttcgttaccc ttttgcaatt caccagcaga agccaacatt
4861 ctctttctac cgttttccaa ttcgaacaaa gagtactttg gcaacttaat aatcaagtcc
4921 ttcttaactt ccttgtaacc cttagcttcc aagaagtcaa ttgggttctt ttcgaaagaa
4981 gatctttcca taatggtaat acccaacaat tccttaacag acttcaactt cttagactta
5041 cccttttcaa ccttagcaac aaccaaaaca gagtaagcaa cggttggaga gtcgaaacca
5101 ccgtacttct ttgggtccca gtccttcttt ctagcaatca acttgtcaga gtttctcttt
5161 ggcaaaatag attccttaga gaaaccaccg gtttgaactt cggtcttctt aacaatgtta
5221 acttgtggca tagacaaaac ctttctaacg gtagcgaagt ctctaccctt gtcccaaaca
5281 atttcaccgg tttcaccgtt ggtttcaatc aatggtctct ttctaatttc accgttagcc
5341 aaggtaattt cggtcttgaa gaagttcata atgttagagt agaagaagta cttagcggta
5401 gccttaccaa tttcttgttc agacttagca atcatctttc taacgtcgta aaccttgtag
5461 tcaccgtaaa cgaattcaga ttccaacttt gggtacttct taatcaaagc ggtaccaaca
5521 acagcgttca agtaagcgtc gtgagcgtgg tggtagttgt taatttctct aaccttgtag
5581 aattggaagt cctttctgaa gtcagaaacc aacttagact tcaaggtaat aaccttaact
5641 tctctaatca acttgtcgtt ttcgtcgtac ttggtgttca ttctagagtc caaaatttga
5701 gcaacgtgct tggtaatttg tctggtttca accaattgtc tcttaatgaa accagccttg
5761 tccaattcag acaaaccacc tctttcagcc ttggtcaagt tgtcgaactt tctttgggta
5821 atcaacttag cgttcaacaa ttgtctccag tagttcttca tcttcttaac aacttcttca
5881 gatggaacgt tgtcagactt acctctgttc ttgtcagatc tggtcaaaac cttgttgtca
5941 atagagtcgt ccttcaagaa agattgtgga acaatgtggt caacgtcgta gtcagacaat
6001 ctgttaatgt ccaattcttg gtcaacgtac atgtctctac cgttttgcaa gtagtacaag
6061 tacaactttt cgttttgcaa ttgggtgttt tcaactgggt gttccttcaa aatttgagaa
6121 cccaattcct taataccttc ttcaattctc ttcattcttt ctctagagtt cttttgaccc
6181 ttttgggtgg tttggttttc tctagccatt tcaataacaa tgttttctgg cttgtgtcta
6241 cccataacct taaccaattc gtcaacaacc ttaacggttt gcaaaatacc cttcttaata
6301 gctggagaac cagccaagtt agcaatgtgt tcgtgcaaag agtcaccttg accagaaact
6361 tgagcctttt gaatgtcttc cttgaaggtc aaagagtcgt cgtgaatcaa ttgcatgaag
6421 tttctgttag cgaaaccgtc agacttcaag aagtccaaaa tggtcttacc agattgcttg
6481 tctctaatac cgttaatcaa ctttctagac aatctacccc aaccggtgta tcttcttctc
6541 ttcaattgct tcataacctt gtcgtcgaac aagtgagcgt aggtcttcaa tctttcttca
6601 atcatttctc tgtcttcgaa caaggtcaag gtcaaaacaa tgtcttccaa aatgtcttcg
6661 ttttcttcgt tgtccaagaa gtccttgtcc ttaataatct tcaacaagtc gtggtaggta
6721 cccaaagaag cgttgaatct gtcttcaaca ccagaaattt caacagagtc gaaacattca
6781 atcttcttga agtagtcttc cttcaattgc ttaacggtaa cctttctgtt ggtcttgaac
6841 aacaagtcaa caatagcctt cttttgttca ccagacaaga aagctggctt tctcatacct
6901 tcggtaacgt acttaacctt ggtcaattcg ttgtaaacgg tgaagtattc gtacaacaaa
6961 gagtgctttg gcaaaacctt ttcgtttggc aagttcttgt cgaagttggt cattctttca
7021 atgaaagatt gagcagaagc acccttgtca acaacttctt cgaagttcca tggggtaatg
7081 gtttcttcag actttctggt catccaagcg aatctagagt tacctctagc caatggacca
7141 acgtagtatg gaattctgaa ggtcaaaatc ttttcaatct tttctctgtt gtccttcaag
7201 aatgggtaga agtcttcttg tcttctcaaa atagcgtgca attcacccaa gtgaatttgg
7261 tgtggaatag aaccgttgtc gaaggttctt tgctttctca acaagtcttc tctgttcaac
7321 ttaaccaaca attcttcggt accgtccatc ttttccaaaa ttggcttaat gaacttgtag
7381 aattcttctt gagaagcacc accgtcaatg taaccagcgt aaccgttctt agattggtcg
7441 aagaaaattt ccttgtactt ttctggcaat tgttgtctaa ccaaagcctt caacaaggtc
7501 aagtcttggt ggtgttcgtc gtatctctta atcatagaag cagacaatgg agccttggta
7561 atttcggtgt taactctcaa aatgtcagac aacaaaatag cgtcagacaa gttcttagca
7621 gccaagaaca agtcagcgta ttggtcacca atttgagcca acaagttgtc caagtcgtcg
7681 tcgtaggtgt ccttagacaa ttgcaactta gcgtcttcag ccaagtcgaa gttagacttg
7741 aagtttgggg tcaaacccaa agacaaagca atcaagttac cgaacaaacc gttcttcttt
7801 tcacctggca attgagcaat caagttttcc aatcttctag acttagacaa tctagcagac
7861 aaaatagcct tagcgtcaac accagaagcg ttaattgggt tttcttcgaa caattggttg
7921 taggtttgaa ccaattgaat gaacaacttg tcaacgtcag agttgtctgg gttcaagtca
7981 ccttcaatca agaagtgacc tctgaactta atcatgtgag ccaaagccaa gtaaatcaat
8041 ctcaagtcag ccttgtcggt agagtcaacc aacttctttc tcaagtggta aatggttggg
8101 tacttttcgt ggtaagcaac ttcgtcaaca atgttaccga aaattgggtg tctttcgtgc
8161 ttcttgtctt cttcaaccaa gaaagattct tccaatctgt ggaagaaaga gtcgtcaacc
8221 ttagccattt cgttagagaa aatttcttgc aagtaacaaa ttctgttctt tcttctggtg
8281 tatcttcttc tagcggttct cttcaatctg gtagcttcag cggtttcacc agagtcgaac
8341 aacaaagcac caatcaagtt cttcttaata gagtgtctgt cggtgttacc caaaaccttg
8401 aacttcttag atggaacctt gtattcgtcg gtaataacag cccaaccaac agagttggta
8461 ccaatgtcca aaccaataga gtacttcttg tccatatgta ttatcgtatt cctttttcgt
8521 tcacttacta ttgctattcg cactcactac agatctatat tttatgttcc accctgttgt
8581 cagatcagga aaggaatgaa gaggctctat ttaaaccaaa tacacctctt cgtttctcaa
8641 tattgttggg ctttcctttt ttttttcctt ccacgtctat ctattgtctc tatcaaaatt
8701 aggaaaaaga aatgaagaac gatgagccaa acgaggctga gacaatcatg catccgtaca
8761 ttctatgtat gtttgggtgt atctttccag attcggttct tttttttttt cagtccaatt
8821 gaaaaaaaaa aggaacagga acaacgggtc gaaatttgcc gtcacattca gagcgaagag
8881 cgacagggga attggtgggc taaccacctg ttgagcccag ctagcagctg tcggtaaacc
8941 gtcgttgggc ggggcgcgac aatgcagcca gcggaggcat tgtatggttt ccgtctggag
9001 agtgggctta gccggcctgg cactccttct ctaagcagag tgagaaggga atctttttac
9061 tggagaagag tgtttgaatc cagcagaagg taatacgctg cggtattttc tccttacgca
9121 tctgtgcggt atttcacacc gcgttttcga cactggatgg cggcgttagt atcgaatcga
9181 cagcagtata gcgaccagca ttcacatacg attgacgcat gatattactt tctgcgcact
9241 taacttcgca tctgggcaga tgatgtcgag gcgaaaaaaa atataaatca cgctaacatt
9301 tgattaaaat agaacaacta caatataaaa aaactataca aatgacaagt tcttgaaaac
9361 aagaatcttt ttattgtcag tactgattag gggcagggca tgctcatgta gagcgcctgc
9421 tcgccgtccg aggcggtgcc gtcgtacagg gcggtgtcca ggccgcagag ggtgaacccc
9481 atccgccggt acgcgtggat cgccggtgcg ttgacgttgg tgacctccag ccagaggtgc
9541 ccggcgcccc gctcgcgggc gaactccgtc gcgagcccca tcaacgcgcg cccgaccccg
9601 tgcccccggt gctccggggc gacctcgatg tcctcgacgg tcagccggcg gttccagccg
9661 gagtacgaga cgaccacgaa gcccgccagg tcgccgtcgt ccccgtacgc gacgaacgtc
9721 cgggagtccg ggtcgccgtc ctccccgtcg tccgattcgt cgtccgattc gtcgtcgggg
9781 aacaccttgg tcaggggcgg gtccaccggc acctcccgca gggtgaagcc gtccccggtg
9841 gcggtgacgc ggaagacggt gtcggtggtg aaggacccat ccagtgcctc gatggcctcg
9901 gcgtcccccg ggacactggt gcggtaccgg taagccgtgt cgtcaagagt ggtacccatg
9961 gttgtttatg ttcggatgtg atgtgagaac tgtatcctag caagatttta aaaggaagta
10021 tatgaaagaa gaacctcagt ggcaaatcct aaccttttat atttctctac aggggcgcgg
10081 cgtggggaca attcaacgcg tctgtgaggg gagcgtttcc ctgctcgcag gtctgcagcg
10141 aggagccgta atttttgctt cgcgccgtgc ggccatcaaa atgtatggat gcaaatgatt
10201 atacatgggg atgtatgggc taaatgtacg ggcgacagtc acatcatgcc cctgagctgc
10261 gcacgtcaag actgtcaagg agggtattct gggcctccat gtcgctgatt accctgttat
10321 cccta
//
