| 质粒名称: | BB3cN_pGAP_23*_pLAT1_Cas9 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 104906 |
| 存储实验室: | Brigitte Gasser |
|
载体骨架基本信息 |
|
| 载体名称: | Episomal ARS/CEN Backbone |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 8867bp |
| 载体类型: | Yeast Expression, CRISPR |
| 筛选标记: | NTC |
| 载体抗性: | Nourseothricin (clonNat), 50 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名:1: | HH - FS23 linker - HDV |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BbsI (not destroyed) |
| 3’克隆位点: | BbsI (not destroyed) |
| 5’测序引物 |
pGAP fw
(Common Sequencing Primers)
|
| 插入基因名称2： | hcas9、human codon-optimized hcas9 sequence fused to a nuclear localization signal (NLS) |
| 物种来源： | H. sapiens (human); Streptococcus pyogenes, Pichia pastoris |
| 插入大小： | (bp)4140 |
|
文献引用方法 |
|
| 材料和方法部分: | BB3cN_pGAP_23*_pLAT1_Cas9 was a gift from Brigitte Gasser (Addgene plasmid # 104906 ; http://n2t.net/addgene:104906 ; RRID:Addgene_104906) |
| 参考文献部分: | CRISPR/Cas9-Mediated Homology-Directed Genome Editing in Pichia pastoris. Gassler T, Heistinger L, Mattanovich D, Gasser B, Prielhofer R. Methods Mol Biol. 2019;1923:211-225. doi: 10.1007/978-1-4939-9024-5_9. 10.1007/978-1-4939-9024-5_9 PubMed 30737742 |
买家导航
BB3cN_pGAP_23*_pLAT1_Cas9载体质粒基本信息
BB3cN_pGAP_23*_pLAT1_Cas9质粒图谱载体图谱和BB3cN_pGAP_23*_pLAT1_Cas9载体序列质粒序列多克隆位点信息
BB3cN_pGAP_23*_pLAT1_Cas9质粒载体简介
BB3cN_pGAP_23*_pLAT1_Cas9质粒序列载体序列
LOCUS BB3cN_pGAP_23*_pLAT1_Cas9 8867 bp DNA circular SYN 14-APR-2025
DEFINITION hCas9 under control of LAT1 for direct cloning of HH-sgRNA-HDV PCR
products for episomal expression in P. pastoris and selection on
NTC.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8867)
AUTHORS Gassler T, Heistinger L, Mattanovich D, Gasser B, Prielhofer R
TITLE CRISPR/Cas9-Mediated Homology-Directed Genome Editing in Pichia
pastoris.
JOURNAL Methods Mol Biol. 2019;1923:211-225. doi:
10.1007/978-1-4939-9024-5_9.
PUBMED 30737742
REFERENCE 2 (bases 1 to 8867)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Methods Mol
Biol."; date: "2019"; volume: "1923:211-225. doi"; pages: "
10.1007/978-1-4939-9024-5_9"
COMMENT Sequence Label: BB3cN_pGAP_23*_pLAT1_Cas9
FEATURES Location/Qualifiers
source 1..8867
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(62..650)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(142..161)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 737..1213
/label=GAP promoter
/note="promoter for Pichia pastoris
glyceraldehyde-3-phosphate dehydrogenase"
CDS 2757..6860
/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 2757..6860
/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 2757..6860
/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 6873..6893
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
terminator 6965..7212
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
primer_bind complement(6981..6999)
/label=CYC1
/note="CYC1 transcription termination signal, reverse
primer"
gene 7232..8351
/label=natMX6
/note="yeast selectable marker conferring nourseothricin
resistance"
promoter 7232..7575
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
CDS 7576..8148
/codon_start=1
/gene="Streptomyces noursei nat1"
/product="nourseothricin acetyltransferase"
/label=NrsR
/note="confers resistance to nourseothricin"
/translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
LCGLDTALYDGTASDGEQALYMSMPCP"
terminator 8154..8351
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
misc_feature 8363..8866
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
ORIGIN
1 cgctgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
61 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
121 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
181 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
241 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
301 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
361 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
421 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
481 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
541 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
601 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
661 tgatcttttc tacggggtgg agcgcaaaaa accccgcccc tgacagggcg gggttttttc
721 gcgatcggag gatccttttt tgtagaaatg tcttggtgtc ctcgtccaat caggtagcca
781 tctctgaaat atctggctcc gttgcaactc cgaacgacct gctggcaacg taaaattctc
841 cggggtaaaa cttaaatgtg gagtaatgga accagaaacg tctcttccct tctctctcct
901 tccaccgccc gttaccgtcc ctaggaaatt ttactctgct ggagagcttc ttctacggcc
961 cccttgcagc aatgctcttc ccagcattac gttgcgggta aaacggaggt cgtgtacccg
1021 acctagcagc ccagggatgg aaaagtcccg gccgtcgctg gcaataatag cgggcggacg
1081 catgtcatga gattattgga aaccaccaga atcgaatata aaaggcgaac acctttccca
1141 attttggttt ctcctgaccc aaagacttta aatttaattt atttgtccct atttcaatca
1201 attgaacaac tatcaaaaca ccatggcgtc ttcggatccc tcgaggaaga cgcgctttta
1261 gtgtacatct gataatatag tactaccacg tatgataatg tagagaatag gcttccttgt
1321 cgagtgtgtt tgcagttttc ttgagtttca aggtttaaat gctggtatat tagttcatcg
1381 aaggtttcag ccaatagcac cttaaatcaa tcaaactaat tcgactctta cgaaagagcc
1441 tactgtgttt agtatcgaag tcgtttacct ttcatgttga atagcttcct ctctgaccct
1501 aacatttcaa gatcctccta aagttacccg gattgtgaaa ttctaatgat ccacctgccc
1561 aatgcatttt ttctttattc agtttacctt ttttacctaa tatacgagct tgttaaagta
1621 agtggcactg caatactagg cttattgttg atattatgat gaatcgtttt cacaaacttg
1681 atttcctgtg aactcaccat gtactaagga aaaaaacatg catcaccatc tgaatatttg
1741 accgctccgg ggagggaaga atatgaacca cgattgaaag actttgggtt tgccgattct
1801 aaaacactta ctgaacagaa aagaagtgaa ttattagaga gtttatgcaa cggagatcat
1861 gagcttagaa agaacgtagg ctgggcaaca acatccattt ctgcctgcga tatttccagt
1921 gcaatgctaa agccacataa tataggcaat tacaacttga atgagcaggc tcatgatgtc
1981 accatggaac tgatagataa agtcatccaa aaaggtgtca atttacggca cgtctacgta
2041 gatactgtcg gacctccagc atcataccaa aagaaactgc aacaaaggtt tcccgcactg
2101 ctgatcaccg tatctaaaaa agcagatgcc atctatccat gtgtttcaag tgcttcagtt
2161 tgcgccaagg tcactagaga cgtctgtttg agaacctcaa atggggagga gaacctatta
2221 gattgtgggt ctggatatcc ctcggaccca aatactaaga aatggctagc ccagtctata
2281 catccattgt ttggctggag tcgtgatgtc aggttttcgt ggcagacggc caaagattgt
2341 cttgagagac acaactcaat caaagttgtg tttggcgaag aggagcctac cccggttaat
2401 ttcggtgatc tggggttgag caaacagaag cccgacttta aggatttaga aatcagtacc
2461 agttggtggg gacattaatt ggtggaggag agccaatatt ctcaactgtt cgtaggacca
2521 taacccccca ctcgcaccag aaatactcat tggcgctgac aactagtttg agccgcttca
2581 attgcggcaa gtccccagtt ttgtttggta ctcccctctc gcctcgcgtg tttttgttcc
2641 tccgaattgt gcatcaccct gtcacatccg tacaccatcg gaacactctg ctcctcctcc
2701 tttctcctct taattgaacg tatatctaat taccctaata acaagtatct tcaaccatgg
2761 acaagaagta ctccattggg ctcgatatcg gcacaaacag cgtcggttgg gccgtcatta
2821 cggacgagta caaggtgccg agcaaaaaat tcaaagttct gggcaatacc gatcgccaca
2881 gcataaagaa gaacctcatt ggcgccctcc tgttcgactc cggggagacg gccgaagcca
2941 cgcggctcaa aagaacagca cggcgcagat atacccgcag aaagaatcgg atctgctacc
3001 tgcaggagat ctttagtaat gagatggcta aggtggatga ctctttcttc cataggctgg
3061 aggagtcctt tttggtggag gaggataaaa agcacgagcg ccacccaatc tttggcaata
3121 tcgtggacga ggtggcgtac catgaaaagt acccaaccat atatcatctg aggaagaagc
3181 ttgtagacag tactgataag gctgacttgc ggttgatcta tctcgcgctg gcgcatatga
3241 tcaaatttcg gggacacttc ctcatcgagg gggacctgaa cccagacaac agcgatgtcg
3301 acaaactctt tatccaactg gttcagactt acaatcagct tttcgaagag aacccgatca
3361 acgcatccgg agttgacgcc aaagcaatcc tgagcgctag gctgtccaaa tcccggcggc
3421 tcgaaaacct catcgcacag ctccctgggg agaagaagaa cggcctgttt ggtaatctta
3481 tcgccctgtc actcgggctg acccccaact ttaaatctaa cttcgacctg gccgaagatg
3541 ccaagcttca actgagcaaa gacacctacg atgatgatct cgacaatctg ctggcccaga
3601 tcggcgacca gtacgcagac ctttttttgg cggcaaagaa cctgtcagac gccattctgc
3661 tgagtgatat tctgcgagtg aacacggaga tcaccaaagc tccgctgagc gctagtatga
3721 tcaagcgcta tgatgagcac caccaagact tgactttgct gaaggccctt gtcagacagc
3781 aactgcctga gaagtacaag gaaattttct tcgatcagtc taaaaatggc tacgccggat
3841 acattgacgg cggagcaagc caggaggaat tttacaaatt tattaagccc atcttggaaa
3901 aaatggacgg caccgaggag ctgctggtaa agcttaacag agaagatctg ttgcgcaaac
3961 agcgcacttt cgacaatgga agcatccccc accagattca cctgggcgaa ctgcacgcta
4021 tcctcaggcg gcaagaggat ttctacccct ttttgaaaga taacagggaa aagattgaga
4081 aaatcctcac atttcggata ccctactatg taggccccct cgcccgggga aattccagat
4141 tcgcgtggat gactcgcaaa tcagaagaga ctatcactcc ctggaacttc gaggaagtcg
4201 tggataaggg ggcctctgcc cagtccttca tcgaaaggat gactaacttt gataaaaatc
4261 tgcctaacga aaaggtgctt cctaaacact ctctgctgta cgagtacttc acagtttata
4321 acgagctcac caaggtcaaa tacgtcacag aagggatgag aaagccagca ttcctgtctg
4381 gagagcagaa gaaagctatc gtggacctcc tcttcaagac gaaccggaaa gttaccgtga
4441 aacagctcaa agaggactat ttcaaaaaga ttgaatgttt cgactctgtt gaaatcagcg
4501 gagtggagga tcgcttcaac gcatccctgg gaacgtatca cgatctcctg aaaatcatta
4561 aagacaagga cttcctggac aatgaggaga acgaggacat tcttgaggac attgtcctca
4621 cccttacgtt gtttgaagat agggagatga ttgaagaacg cttgaaaact tacgctcatc
4681 tcttcgacga caaagtcatg aaacagctca agaggcgccg atatacagga tgggggcggc
4741 tgtcaagaaa actgatcaat gggatccgag acaagcagag tggaaagaca atcctggatt
4801 ttcttaagtc cgatggattt gccaaccgga acttcatgca gttgatccat gatgactctc
4861 tcacctttaa ggaggacatc cagaaagcac aagtttctgg ccagggggac agccttcacg
4921 agcacatcgc taatcttgca ggtagcccag ctatcaaaaa gggaatactg cagaccgtta
4981 aggtcgtgga tgaactcgtc aaagtaatgg gaaggcataa gcccgagaat atcgttatcg
5041 agatggcccg agagaaccaa actacccaga agggacagaa gaacagtagg gaaaggatga
5101 agaggattga agagggtata aaagaactgg ggtcccaaat ccttaaggaa cacccagttg
5161 aaaacaccca gcttcagaat gagaagctct acctgtacta cctgcagaac ggcagggaca
5221 tgtacgtgga tcaggaactg gacatcaatc ggctctccga ctacgacgtg gatcatatcg
5281 tgccccagtc ttttctcaaa gatgattcta ttgataataa agtgttgaca agatccgata
5341 aaaatagagg gaagagtgat aacgtcccct cagaagaagt tgtcaagaaa atgaaaaatt
5401 attggcggca gctgctgaac gccaaactga tcacacaacg gaagttcgat aatctgacta
5461 aggctgaacg aggtggcctg tctgagttgg ataaagccgg cttcatcaaa aggcagcttg
5521 ttgagacacg ccagatcacc aagcacgtgg cccaaattct cgattcacgc atgaacacca
5581 agtacgatga aaatgacaaa ctgattcgag aggtgaaagt tattactctg aagtctaagc
5641 tcgtctcaga tttcagaaag gactttcagt tttataaggt gagagagatc aacaattacc
5701 accatgcgca tgatgcctac ctgaatgcag tggtaggcac tgcacttatc aaaaaatatc
5761 ccaagcttga atctgaattt gtttacggag actataaagt gtacgatgtt aggaaaatga
5821 tcgcaaagtc tgagcaggaa ataggcaagg ccaccgctaa gtacttcttt tacagcaata
5881 ttatgaattt tttcaagacc gagattacac tggccaatgg agagattcgg aagcgaccac
5941 ttatcgaaac aaacggagaa acaggagaaa tcgtgtggga caagggtagg gatttcgcga
6001 cagtccggaa ggtcctgtcc atgccgcagg tgaacatcgt taaaaagacc gaagtacaga
6061 ccggaggctt ctccaaggaa agtatcctcc cgaaaaggaa cagcgacaag ctgatcgcac
6121 gcaaaaaaga ttgggacccc aagaaatacg gcggattcga ttctcctaca gtcgcttaca
6181 gtgtactggt tgtggccaaa gtggagaaag ggaagtctaa aaaactcaaa agcgtcaagg
6241 aactgctggg catcacaatc atggagcgat caagcttcga aaaaaacccc atcgactttc
6301 tcgaggcgaa aggatataaa gaggtcaaaa aagacctcat cattaagctt cccaagtact
6361 ctctctttga gcttgaaaac ggccggaaac gaatgctcgc tagtgcgggc gagctgcaga
6421 aaggtaacga gctggcactg ccctctaaat acgttaattt cttgtatctg gccagccact
6481 atgaaaagct caaagggtcc cccgaagata atgagcagaa gcagctgttc gtggaacaac
6541 acaaacacta ccttgatgag atcatcgagc aaataagcga attctccaaa agagtgatcc
6601 tcgccgacgc taacctcgat aaggtgcttt ctgcttacaa taagcacagg gataagccca
6661 tcagggagca ggcagaaaac attatccact tgtttactct gaccaacttg ggcgcgcctg
6721 cagccttcaa gtacttcgac accaccatag acagaaagcg gtacacctct acaaaggagg
6781 tcctggacgc cacactgatt catcagtcaa ttacggggct ctatgaaaca agaatcgacc
6841 tctctcagct cggtggagac agcagggctg accccaagaa gaagaggaag gtgtgagctt
6901 cgtccgacgg cggcccgacg ggtccgaggc ctcggagatc cgtccccctt ttcctttgtc
6961 gatatcatgt aattagttat gtcacgctta cattcacgcc ctccccccac atccgctcta
7021 accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt ttttatagtt
7081 atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct gtacagacgc
7141 gtgtacgcat gtaacattat actgaaaacc ttgcttgaga aggttttggg acgctcgaag
7201 gctttaattt gcaagctggc gctaattcat ggacatggag gcccagaata ccctccttga
7261 cagtcttgac gtgcgcagct caggggcatg atgtgactgt cgcccgtaca tttagcccat
7321 acatccccat gtataatcat ttgcatccat acattttgat ggccgcacgg cgcgaagcaa
7381 aaattacggc tcctcgctgc agacctgcga gcagggaaac gctcccctca cagacgcgtt
7441 gaattgtccc cacgccgcgc ccctgtagag aaatataaaa ggttaggatt tgccactgag
7501 gttcttcttt catatacttc cttttaaaat cttgctagga tacagttctc acatcacatc
7561 cgaacataaa caaccatggg taccactctt gacgacacgg cttaccggta ccgcaccagt
7621 gtcccggggg acgccgaggc catcgaggca ctggatgggt ccttcaccac cgacaccgtt
7681 tttcgcgtca ccgccaccgg ggacggcttc accctgcggg aggtgccggt ggacccgccc
7741 ctgaccaagg tgttccccga cgacgaatcg gacgacgaat cggacgacgg ggaggacggc
7801 gacccggact cccggacgtt cgtcgcgtac ggggacgacg gcgacctggc gggcttcgtg
7861 gtcgtctcgt actccggctg gaaccgccgg ctgaccgtcg aggacatcga ggtcgccccg
7921 gagcaccggg ggcacggggt cgggcgcgcg ttgatggggc tcgcgacgga gttcgcccgc
7981 gagcggggcg ccgggcacct ctggctggag gtcaccaacg tcaacgcacc ggcgatccac
8041 gcgtaccggc ggatggggtt caccctctgc ggcctggaca ccgccctgta cgacggcacc
8101 gcctcggacg gcgagcaggc gctctacatg agcatgccct gcccctaatc agtactgaca
8161 ataaaaagat tcttgttttc aagaacttgt catttgtata gtttttttat attgtagttg
8221 ttctatttta atcaaatgtt agcgtgattt atattttttt tcgcctcgac atcatctgcc
8281 cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt caatcgtatg tgaatgctgg
8341 tcgctatact ggcttccttt tcatcacgtg ctataaaaat aattataatt taaatttttt
8401 aatataaata tataaattaa aaatagaaag taaaaaaaga aattaaagaa aaaatagttt
8461 ttgttttccg aagatgtaaa agactctagg gggatcgcca acaaatacta ccttttatct
8521 tgctcttcct gctctcaggt attaatgccg aattgtttca tcttgtctgt gtagaacacc
8581 acacacgaaa atcctgtgat tttacatttt acttatcgtt aatcgaatgt atatctattt
8641 aatctgcttt tcttgtctaa taaatatata tgtaaagtac gctttttgtt gaaatttttt
8701 aaacctttgt ttattttttt ttcttcattc cgtaactctt ctaccttctt tatttacttt
8761 ctaaaatcca aatacaaaac ataaaaataa ataaacacag agtaaattcc caaattattc
8821 catcattaaa agatacgagg cgcgtgtaag ttacaggcaa gcgatcg
//
