| 质粒名称: | pSAG1::CAS9-U6::sgUPRT |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 54467 |
| 存储实验室: | David Sibley |
|
载体骨架基本信息 |
|
| 载体名称: | pBlueScript |
| 载体出品公司: | Agilent |
| 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: | CRISPR,弓形虫表达 |
| 筛选标记: | -- |
| 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: | SAG1 5' UTR |
| 物种来源: | Toxoplasma gondii |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 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: | pSAG1::CAS9-U6::sgUPRT was a gift from David Sibley (Addgene plasmid # 54467 ; http://n2t.net/addgene:54467 ; RRID:Addgene_54467) |
| 参考文献部分: | Efficient Gene Disruption in Diverse Strains of Toxoplasma gondii Using CRISPR/CAS9. Shen B, Brown KM, Lee TD, Sibley LD. MBio. 2014 May 13;5(3). pii: e01114-14. doi: 10.1128/mBio.01114-14. 10.1128/mBio.01114-14 PubMed 24825012 |
买家导航
pSAG1::CAS9-U6::sgUPRT载体质粒基本信息
pSAG1::CAS9-U6::sgUPRT质粒图谱载体图谱和pSAG1::CAS9-U6::sgUPRT载体序列质粒序列多克隆位点信息
pSAG1::CAS9-U6::sgUPRT质粒载体简介
pSAG1::CAS9-U6::sgUPRT质粒序列载体序列
LOCUS pSAG1::CAS9-U6::sgUPRT 9674 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Encodes CAS9 nuclease (GFP fusion) under control of the Toxoplasma
gondii SAG1 promotor. Vector also provides U6 controlled expression
of a single guide RNA for disruption of genes by CRISPR..
ACCESSION .
VERSION .
KEYWORDS pSAG1::CAS9-U6::sgUPRT
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9674)
AUTHORS Shen B, Brown KM, Lee TD, Sibley LD
TITLE Efficient Gene Disruption in Diverse Strains of Toxoplasma gondii
Using CRISPR/CAS9.
JOURNAL MBio. 2014 May 13;5(3). pii: e01114-14. doi: 10.1128/mBio.01114-14.
PUBMED 24825012
REFERENCE 2 (bases 1 to 9674)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9674
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_RNA 559..633
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter complement(658..676)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(697..713)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(697..713)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(710..732)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 721..737
/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(745..775)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 790..811
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(928..945)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1099..1687)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1179..1198)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1858..2718)
/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 2481..2500
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2719..2823)
/gene="bla"
/label=AmpR promoter
rep_origin complement(2849..3304)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2986..3007)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3198..3217
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3431..3453
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 3445..3462
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 3446..3462
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3472..3491
/label=T7
/note="T7 promoter, forward primer"
promoter 3472..3490
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind 3515..3531
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind 3516..3532
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
CDS 4358..8461
/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"
CDS 8465..8491
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 8510..8530
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 8537..8557
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 8597..9313
/codon_start=1
/product="enhanced GFP with monomerizing A206K mutation
(Zacharias et al., 2002)"
/label=mEGFP
/note="mammalian codon-optimized"
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
primer_bind complement(8639..8660)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind 9247..9268
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(9635..9651)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(9635..9651)
/label=pBluescriptSK
/note="For pBluescript vector"
ORIGIN
1 caagtaagca gaagcacgct gtatttccgg gagggtgcga cgagacaaag tgcgcgagtt
61 gaaatcgtcg tggggacgat tgcaccgcgg ccacatgttg gagacactga ggacacacgg
121 gaaacgcgaa agatttcaaa ttaacgtacc caagcgcgaa agcttgcgca gcatacactc
181 gaagcgaaca tcccgaacca tcgagaggcg gagagcgata agtctttcac gctgcgaagt
241 gttgcgacgg ctgcgccgct gcactgtgaa ttgggcgcca atattgcatc ctaggcctga
301 cgcgcctcct gcagaacgcg agacactggg atatgtagag ccaaggggga aaccttcgaa
361 ctctcgaatg tcttctctga caagaatcat atttccatca gttctgtcag attttcaaat
421 ggcgacctgc agaggcctgc ttcctccctg tgcgctcttc gaaggggctt tctgtcgcgc
481 agggtcacct cgtccccgaa gggggtgttt gccttctggt aaatggggat gtcaagttgg
541 cgtctcgatt gtgagagcgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg
601 ttatcaactt gaaaagtggc accgagtcgg tgcttttttg agctccagct tttgttccct
661 ttagtgaggg ttaatttcga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
721 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
781 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
841 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
901 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
961 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
1021 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
1081 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
1141 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
1201 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
1261 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
1321 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
1381 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
1441 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
1501 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
1561 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
1621 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
1681 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
1741 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
1801 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
1861 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
1921 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
1981 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
2041 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
2101 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
2161 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
2221 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
2281 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
2341 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
2401 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
2461 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
2521 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
2581 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
2641 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
2701 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
2761 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
2821 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc
2881 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
2941 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
3001 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
3061 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
3121 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
3181 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
3241 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
3301 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
3361 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
3421 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac
3481 tcactatagg gcgaattggg taccgggccc cccctcgagg tcgacggtat cgataagctt
3541 ttacatccgt tgccttttcc acggtccgtg atttcatgtg cgtgcagctt caaagactgg
3601 tcgttgcgac taataagact gcagtgacag gtcgaatggt gggcaccttg ctgatgacta
3661 tctactgcaa agtctgagac aacgaacgaa acttcccaca cgaggcattt gaaactgacg
3721 gtgtctaggt aatatgcact gcaagacacg gtactggggc ctcgctgaat taggggccga
3781 tctcgttgcc ctatcagtgc tcacagtgcc gcaacgtaac accagggcag gttcttgaca
3841 gtggcaacaa tgtgcgacgg gcgtgtgaac gtttcgtagt catagcgcta gcacgtacct
3901 agccacatgg tcgtgaggag ctttaccatg cgtctagaag gtggatgcgg gacacgcctt
3961 cctggccttt ggctcccgag acgcgtgttc taaccacaaa ccttgagacg cgtgttccaa
4021 ccacgcaccc tgacacgcgt gttccaacca cgcaccctga gacgcgtgtt ctaaccacgc
4081 accctgagac gcgtgttcta accacgcacc ctgagacgcg tgttctgccg cacaatgtgc
4141 acctgtagga agctgtagtc actgctgatt ctcactgttc tcggcaaggg ccgacgaccg
4201 gagtacagtt tttgtgggca gagccgttgt gcagctttcc gttcttctcg gttgtgtcac
4261 atgtgtcatt gtcgtgtaaa cacacggttg tatgtcggtt tcgctgcacc acttcattat
4321 ttcttctggt tttttgacga gtatgcatct agacaaaatg gacaagaagt acagcatcgg
4381 cctggacatc ggcaccaact ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc
4441 cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga agaacctgat
4501 cggcgccctg ctgttcgaca gcggagaaac agccgaggcc acccggctga agagaaccgc
4561 cagaagaaga tacaccagac ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa
4621 cgagatggcc aaggtggacg acagcttctt ccacagactg gaagagtcct tcctggtgga
4681 agaggataag aagcacgagc ggcaccccat cttcggcaac atcgtggacg aggtggccta
4741 ccacgagaag taccccacca tctaccacct gagaaagaaa ctggtggaca gcaccgacaa
4801 ggccgacctg cggctgatct atctggccct ggcccacatg atcaagttcc ggggccactt
4861 cctgatcgag ggcgacctga accccgacaa cagcgacgtg gacaagctgt tcatccagct
4921 ggtgcagacc tacaaccagc tgttcgagga aaaccccatc aacgccagcg gcgtggacgc
4981 caaggccatc ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc tgatcgccca
5041 gctgcccggc gagaagaaga atggcctgtt cggcaacctg attgccctga gcctgggcct
5101 gacccccaac ttcaagagca acttcgacct ggccgaggat gccaaactgc agctgagcaa
5161 ggacacctac gacgacgacc tggacaacct gctggcccag atcggcgacc agtacgccga
5221 cctgtttctg gccgccaaga acctgtccga cgccatcctg ctgagcgaca tcctgagagt
5281 gaacaccgag atcaccaagg cccccctgag cgcctctatg atcaagagat acgacgagca
5341 ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg agaagtacaa
5401 agagattttc ttcgaccaga gcaagaacgg ctacgccggc tacatcgatg gcggagccag
5461 ccaggaagag ttctacaagt tcatcaagcc catcctggaa aagatggacg gcaccgagga
5521 actgctcgtg aagctgaaca gagaggacct gctgcggaag cagcggacct tcgacaacgg
5581 cagcatcccc caccagatcc acctgggaga gctgcacgcc attctgcggc ggcaggaaga
5641 tttttaccca ttcctgaagg acaaccggga aaagatcgag aagatcctga ccttccgcat
5701 cccctactac gtgggccctc tggccagggg aaacagcaga ttcgcctgga tgaccagaaa
5761 gagcgaggaa accatcaccc cctggaactt cgaggaagtg gtggacaagg gcgccagcgc
5821 ccagagcttc atcgagcgga tgaccaactt cgataagaac ctgcccaacg agaaggtgct
5881 gcccaagcac agcctgctgt acgagtactt caccgtgtac aacgagctga ccaaagtgaa
5941 atacgtgacc gagggaatga gaaagcccgc cttcctgagc ggcgagcaga aaaaagccat
6001 cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga aagaggacta
6061 cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa
6121 cgcctccctg ggcacatacc acgatctgct gaaaattatc aaggacaagg acttcctgga
6181 caatgaggaa aacgaggaca ttctggaaga tatcgtgctg accctgacac tgtttgagga
6241 cagagagatg atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat
6301 gaagcagctg aagcggcgga gatacaccgg ctggggcagg ctgagccgga agctgatcaa
6361 cggcatccgg gacaagcagt ccggcaagac aatcctggat ttcctgaagt ccgacggctt
6421 cgccaacaga aacttcatgc agctgatcca cgacgacagc ctgaccttta aagaggacat
6481 ccagaaagcc caggtgtccg gccagggcga tagcctgcac gagcacattg ccaatctggc
6541 cggcagcccc gccattaaga agggcatcct gcagacagtg aaggtggtgg acgagctcgt
6601 gaaagtgatg ggccggcaca agcccgagaa catcgtgatc gaaatggcca gagagaacca
6661 gaccacccag aagggacaga agaacagccg cgagagaatg aagcggatcg aagagggcat
6721 caaagagctg ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa
6781 cgagaagctg tacctgtact acctgcagaa tgggcgggat atgtacgtgg accaggaact
6841 ggacatcaac cggctgtccg actacgatgt ggaccatatc gtgcctcaga gctttctgaa
6901 ggacgactcc atcgataaca aagtgctgac tcggagcgac aagaaccggg gcaagagcga
6961 caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac tactggcgcc agctgctgaa
7021 tgccaagctg attacccaga ggaagttcga caatctgacc aaggccgaga gaggcggcct
7081 gagcgaactg gataaggccg gcttcatcaa gagacagctg gtggaaaccc ggcagatcac
7141 aaagcacgtg gcacagatcc tggactcccg gatgaacact aagtacgacg agaacgacaa
7201 actgatccgg gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg atttccggaa
7261 ggatttccag ttttacaaag tgcgcgagat caacaactac caccacgccc acgacgccta
7321 cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg aaagcgagtt
7381 cgtgtacggc gactacaagg tgtacgacgt gcggaagatg atcgccaaga gcgagcagga
7441 aatcggcaag gctaccgcca agtacttctt ctacagcaac atcatgaact ttttcaagac
7501 cgagattacc ctggccaacg gcgagatccg gaagcggcct ctgatcgaga caaacggcga
7561 aacaggcgag atcgtgtggg ataagggccg ggactttgcc accgtgcgga aagtgctgtc
7621 tatgccccaa gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga
7681 gtctatcctg cccaagagga acagcgacaa gctgatcgcc agaaagaagg actgggaccc
7741 taagaagtac ggcggcttcg acagccccac cgtggcctat tctgtgctgg tggtggccaa
7801 agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat
7861 catggaaaga agcagcttcg agaagaatcc catcgacttt ctggaagcca agggctacaa
7921 agaagtgaaa aaggacctga tcatcaagct gcctaagtac tccctgttcg agctggaaaa
7981 cggccggaag agaatgctgg cctctgccgg cgaactgcag aagggaaacg aactggccct
8041 gccctccaaa tatgtgaact tcctgtacct ggccagccac tatgagaagc tgaagggctc
8101 ccccgaggat aatgagcaga aacagctgtt tgtggaacag cacaaacact acctggacga
8161 gatcatcgag cagatcagcg agttctccaa gagagtgatc ctggccgacg ctaatctgga
8221 caaggtgctg agcgcctaca acaagcacag agacaagcct atcagagagc aggccgagaa
8281 tatcatccac ctgtttaccc tgaccaatct gggagcccct gccgccttca agtactttga
8341 caccaccatc gaccggaaga ggtacaccag caccaaagag gtgctggacg ccaccctgat
8401 ccaccagagc atcaccggcc tgtacgagac acggatcgac ctgtctcagc tgggaggcga
8461 cgcctatccc tatgacgtgc ccgattatgc cagcctgggc agcggctccc ccaagaaaaa
8521 acgcaaggtg gaagatccta agaaaaagcg gaaagtggac ggcattggta gtgggagcaa
8581 cggcagcagc ggatccgtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
8641 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgcgcggcg agggcgaggg
8701 cgatgccacc aacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
8761 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
8821 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
8881 gcgcaccatc tccttcaagg acgacggcac ctacaagacc cgcgccgagg tgaagttcga
8941 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
9001 catcctgggg cacaagctgg agtacaactt caacagccac aacgtctata tcacggccga
9061 caagcagaag aacggcatca aggcgaactt caagatccgc cacaacgtcg aggacggcag
9121 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
9181 gcccgacaac cactacctga gcacccagtc caagctgagc aaagacccca acgagaagcg
9241 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
9301 gctgtacaag tagttaatta atcaccgttg tgctcacttc tcaaatcgac aaaggaaaca
9361 cacttcgtgc agcatgtgcc ccattataaa gaaactgagt tgttccgttg tggcttgcag
9421 gtgtcacatc cacaaaaacc ggccgactct aaataggagt gtttcgcagc aagcagcgaa
9481 agtttatgac tgggtccgaa tctctgaacg gatgtgtggc ggacctggct gatgttgatc
9541 gccgtcgaca cacgcgccac atgggtcaat acacaagaca gctatcagtt gttttagtcg
9601 aaccggttaa cacaattctt gcccccccga gggggatcca ctagttctag agcggccgcc
9661 accgcggtgg agct
//
