| 质粒名称: | pPtGE34 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 107932 |
| 存储实验室: | David Edgell |
|
载体骨架基本信息 |
|
| 载体名称: | pKS diaCas9_sgRNA |
| 载体出品公司: | Per Winge |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 8577 bp |
| 完整质粒大小: | 12392 bp |
| 载体修饰: | 添加酵母Cen6-ArsH4-His3、转移来源和P。三角帆蚌 |
| 载体类型: | CRISPR，合成生物学；三角褐指藻的上位体载体 |
| 筛选标记: | 博来霉素,HIS3 |
| 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: | OriT,Origin of Transfer |
| 物种来源: | -- |
| 插入大小: | 771 bp |
| 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: | pPtGE34 was a gift from David Edgell (Addgene plasmid # 107932 ; http://n2t.net/addgene:107932 ; RRID:Addgene_107932) |
| 参考文献部分: | An Expanded Plasmid-Based Genetic Toolbox Enables Cas9 Genome Editing and Stable Maintenance of Synthetic Pathways in Phaeodactylum tricornutum. Slattery SS, Diamond A, Wang H, Therrien JA, Lant JT, Jazey T, Lee K, Klassen Z, Desgagne-Penix I, Karas BJ, Edgell DR. ACS Synth Biol. 2018 Jan 8. doi: 10.1021/acssynbio.7b00191. 10.1021/acssynbio.7b00191 PubMed 29298053 |
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 pPtGE34 12389 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses Cas9 in Phaeodactylum tricornutum / Encodes elements
required for conjugation.
ACCESSION .
VERSION .
KEYWORDS pPtGE34
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12389)
AUTHORS Slattery SS, Diamond A, Wang H, Therrien JA, Lant JT, Jazey T, Lee
K, Klassen Z, Desgagne-Penix I, Karas BJ, Edgell DR
TITLE An Expanded Plasmid-Based Genetic Toolbox Enables Cas9 Genome
Editing and Stable Maintenance of Synthetic Pathways in
Phaeodactylum tricornutum.
JOURNAL ACS Synth Biol. 2018 Jan 8. doi: 10.1021/acssynbio.7b00191.
PUBMED 29298053
REFERENCE 2 (bases 1 to 12389)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..12389
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin complement(3..458)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(140..161)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 352..371
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 600..616
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 623..642
/label=T7
/note="T7 promoter, forward primer"
promoter 623..641
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS complement(782..1153)
/codon_start=1
/gene="traJ"
/product="oriT-recognizing protein"
/label=traJ
/translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
KQDELGKVMMGVVRPRAEP"
oriT 1186..1295
/note="incP origin of transfer"
primer_bind 1572..1588
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 1612..1630
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
CDS 2251..2625
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=BleoR
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFVGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLNELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
promoter 3627..3812
/gene="S. cerevisiae HIS3"
/label=HIS3 promoter
CDS 3813..4469
/codon_start=1
/gene="S. cerevisiae HIS3"
/product="imidazoleglycerol-phosphate dehydratase, required
for histidine biosynthesis"
/label=HIS3
/note="yeast auxotrophic marker"
/translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVL"
CDS 4933..4998
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 5005..5025
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5050..9150
/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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 9151..9198
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
misc_RNA 9734..9809
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
primer_bind complement(10156..10172)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(10157..10173)
/label=pBluescriptKS
/note="For pBluescript vector"
promoter complement(10201..10219)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(10240..10256)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(10240..10256)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(10253..10275)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 10264..10280
/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(10288..10318)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 10333..10354
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(10471..10488)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(10642..11230)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(10722..10741)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(11401..12261)
/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 12024..12043
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(12262..12366)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg agctgatcgt
661 cttgccttgc tcgtcggtga tgtacttcac cagctccgcg aagtcgctct tcttgatgga
721 gcgcatgggg acgtgcttgg caatcacgcg caccccccgg ccgttttagc ggctaaaaaa
781 gtcatggctc tgccctcggg cggaccacgc ccatcatgac cttgccaagc tcgtcctgct
841 tctcttcgat cttcgccagc agggcgagga tcgtggcatc accgaaccgc gccgtgcgcg
901 ggtcgtcggt gagccagagt ttcagcaggc cgcccaggcg gcccaggtcg ccattgatgc
961 gggccagctc gcggacgtgc tcatagtcca cgacgcccgt gattttgtag ccctggccga
1021 cggccagcag gtaggccgac aggctcatgc cggccgccgc cgccttttcc tcaatcgctc
1081 ttcgttcgtc tggaaggcag tacaccttga taggtgggct gcccttcctg gttggcttgg
1141 tttcatcagc catccgcttg ccctcatctg ttacgccggc ggtagccggc cagcctcgca
1201 gagcaggatt cccgttgagc accgccaggt gcgaataagg gacagtgaag aaggaacacc
1261 cgctcgcggg tgggcctact tcacctatcc tgcccggctg acgccgttgg atacaccaag
1321 gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg tgcgaaaaag gatggatata
1381 ccgaaaaaat cgctataatg accccgaagc agggttatgc agcggaagat gccattcgcc
1441 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
1501 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
1561 gtcacgacgt tgtaaaacga cggccagtga ccatgattac gccaagctcg aaattaaccc
1621 tcactaaagg gaacaaaagc tggtacctaa ccctgcgata gaccttttcc aaactcacgc
1681 agtccaagaa aacaaagggg tgagaagtat acgcaccttt cggtttcggc ataattctta
1741 aactcttgtg gtcactttct tgtgaagaag ctaggggcac tcgttttccc tcagagcctg
1801 caaacacaaa attcctgcag tcaattgtcc caacactcgg caagccgtat gcgcaagcaa
1861 cgatgcgcag aaggccgtgg atggatggcg actcgcgata tggcttcttg ggtcgccagt
1921 gtggtatgtc cggcgtatgt caatacgcga attcggacga ctggcatctc taggaggagg
1981 attccttctt ttatgacatg tttattttat atacattgat gctttccgac agtcggaagt
2041 aataaatgaa tttatttcaa gactacctat actcctttga cttgttcgac taatcttacc
2101 gcttactaaa atctcgaaat cacgcttgac ctctcgcacg caaatttttg ctgctggacg
2161 ctacgcactc ggcccaattc ttctcggtcc tcgtcgtcgc aattgtcgtt gcgttgatct
2221 tgcaccgaag gaatcagaga atagaatacc atggccaagt tgaccagtgc cgttccggtg
2281 ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttctcc
2341 cgggacttcg tggaggacga cttcgtcggt gtggtccggg acgacgtgac cctgttcatc
2401 agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc
2461 ctgaacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc
2521 gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac
2581 ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgaagata agaatatctc
2641 attgtgaaca tctatgattt accaatttta ttctttgttt acagttagac gccagtaatt
2701 gtgctgtttc tctcaagtct gtgtcaatac aaactacgaa acttggcaat ttttctcttg
2761 aatatgagca cgagattgaa acgcacaaag gaaattagtt tccatccttt gacaaagttt
2821 gttgctgttt agagaacaga tgtcaaaatt aacgtgccat ggaattgaac catggtgcgc
2881 tatccccaaa tcacgtcgtt tgacctcgtc acatttaagg tatatcaagc atattcactt
2941 atatcttgac atcctccctg cttgatattc ctttgccctt gagccatctt ccccacacgc
3001 tggagatgac ccctgcccct cctttcttta tccacccgag taaggtgtcg acaagtcact
3061 ttgcccctga gccatcctcc ccactcagga gatgacccct acccctttct tctttatcca
3121 cccccacgtg ctataaaaat aattataatt taaatttttt aatataaata tataaattaa
3181 aaatagaaag taaaaaaaga aattaaagaa aaaatagttt ttgttttccg aagatgtaaa
3241 agactctagg gggatcgcca acaaatacta ccttttacct tgctcttcct gctctcaggt
3301 attaatgccg aattgtttca tcttgtctgt gtagaagacc acacacgaaa atcctgtgat
3361 tttacatttt acttatcgtt aatcgaatgt atatctattt aatctgcttt tcttgtctaa
3421 taaatatata tgtaaagtac gctttttgtt gaaatttttt aaacctttgt ttattttttt
3481 ttcttcattc cgtaactctt ctaccttctt tatttacttt ctaaaatcca aatacaaaac
3541 ataaaaataa ataaacacag agtaaattcc caaattattc catcattaaa agatacgagg
3601 cgcgtgtaag ttacaggcaa gcgatcctag tacactctat atttttttat gcctcggtaa
3661 tgattttcat ttttttttcc acctagcgga tgactctttt tttttcttag cgattggcat
3721 tatcacataa tgaattatac attatataaa gtaatgtgat ttcttcgaag aatatactaa
3781 aaaatgagca ggcaagataa acgaaggcaa agatgacaga gcagaaagcc ctagtaaagc
3841 gtattacaaa tgaaaccaag attcagattg cgatctcttt aaagggtggt cccctagcga
3901 tagagcactc gatcttccca gaaaaagagg cagaagcagt agcagaacag gccacacaat
3961 cgcaagtgat taacgtccac acaggtatag ggtttctgga ccatatgata catgctctgg
4021 ccaagcattc cggctggtcg ctaatcgttg agtgcattgg tgacttacac atagacgacc
4081 atcacaccac tgaagactgc gggattgctc tcggtcaagc ttttaaagag gccctagggg
4141 ccgtgcgtgg agtaaaaagg tttggatcag gatttgcgcc tttggatgag gcactttcca
4201 gagcggtggt agatctttcg aacaggccgt acgcagttgt cgaacttggt ttgcaaaggg
4261 agaaagtagg agatctctct tgcgagatga tcccgcattt tcttgaaagc tttgcagagg
4321 ctagcagaat taccctccac gttgattgtc tgcgaggcaa gaatgatcat caccgtagtg
4381 agagtgcgtt caaggctctt gcggttgcca taagagaagc cacctcgccc aatggtacca
4441 acgatgttcc ctccaccaaa ggtgttcttt atgcaatctc gcctattcat ggtgtataaa
4501 agttcaacat ccaaagctag aacttttgga aagagaaaga atatccgaat agggcacggc
4561 gtgccgtatt gttggagtgg actagcagaa agtgaggaag gcacaggatg agttttctcg
4621 agacacatag cttcagcgtc gtgtaggcta ggcagaggtg agttttctcg agacatacct
4681 tcagcgtcgt cttcactgtc acagtcaact gacagtaatc gttgatccgg agagattcaa
4741 aattcaatct gtttggacct ggataagaca caagagcgac atcctgacat gaacgccgta
4801 aacagcaaat cctggttgaa cacgtatcct tttgggggcc tccgctacga cgctcgctcc
4861 agctggggct tccttactat acacagcgcg catatttcac ggttgccaga agtcaagatg
4921 gcctctagaa tggactataa ggaccacgac ggagattaca aggatcatga tattgattac
4981 aaagacgatg atgataaaat ggccccaaag aagaagcgga aagtcggtat acacggagtc
5041 ccagcagccg acaagaagta ctcgattgga ttggacatcg gcaccaactc tgttggatgg
5101 gccgtcatca cggacgagta caaggtgccc agcaagaaat tcaaagtact gggaaacact
5161 gaccgccatt cgatcaagaa gaatttgatt ggagctttgc tgtttgacag cggcgaaaca
5221 gcggaagcca cgcggcttaa acgaaccgcg agacgaagat atacgcgacg gaagaaccgt
5281 atctgctatc tacaagagat tttctcgaat gagatggcaa aagtcgatga ctctttcttt
5341 caccgattgg aagaatcctt cctcgttgaa gaggataaga aacatgagcg gcacccgata
5401 tttggaaaca ttgtggacga ggtggcttac catgagaaat acccgactat ttaccacctg
5461 cgaaagaaat tggtggactc gacggacaaa gcggatcttc gcctaatcta tttggcgcta
5521 gcacatatga ttaagtttcg cggccacttc ttgatcgaag gcgacttgaa tccggacaac
5581 agtgatgttg acaaactgtt tattcaactt gtacaaactt acaaccagct ctttgaagaa
5641 aatcccatta acgcttcggg agtggacgcg aaagccatcc tttctgcccg actcagcaag
5701 agccgacgcc tagaaaattt gattgcgcaa cttcccggcg agaaaaagaa tggacttttt
5761 ggaaatctta ttgctttgtc gctgggcctt acgccgaact ttaagagcaa cttcgacctt
5821 gcggaagatg ccaaattgca actgagtaaa gacacatatg acgacgacct cgacaatctt
5881 ttggcacaaa taggagatca gtacgccgac ctctttttgg cggctaagaa cttgtccgac
5941 gccattctgt tgtcggacat ccttcgagtt aatacggaga ttactaaggc gcccttgagc
6001 gcctctatga tcaaaagata cgacgagcat caccaagacc tgaccctcct taaagctctc
6061 gtccggcaac agctgcctga gaagtacaaa gagattttct ttgatcaaag caaaaacgga
6121 tacgcgggct acattgacgg gggagcctcg caagaagagt tctacaagtt tattaaaccg
6181 atcctggaaa agatggacgg cacggaggaa ttactcgtga aacttaaccg agaggacctt
6241 ttgcgtaagc aacggacttt cgacaacgga tctattccgc accaaatcca tctcggagaa
6301 ttgcacgcaa ttctgcgccg gcaagaagat ttttatccat ttctcaagga caaccgggaa
6361 aagattgaga aaatcttaac cttccgcata ccctactacg tcggcccttt ggccagggga
6421 aattcgagat ttgcctggat gacgcgaaag agtgaggaaa ctattactcc ctggaatttc
6481 gaagaagtgg tagacaaagg agcttccgcg caatccttta tcgagcgcat gaccaacttc
6541 gataagaacc tcccgaatga gaaagtccta cccaagcatt cgttgctcta cgaatacttt
6601 acggtgtata atgagctgac taaagtcaaa tacgttacag agggaatgcg aaaaccggct
6661 ttcctctcgg gagagcagaa aaaggccata gtcgatcttc tctttaaaac gaaccgtaaa
6721 gttaccgtga aacaactaaa agaggactac ttcaagaaaa tcgaatgctt tgactccgtt
6781 gaaatttccg gcgttgaaga tcggttcaat gcctccttgg gaacatacca cgatctcctg
6841 aaaattatca aagacaagga ctttcttgac aatgaggaaa acgaggacat tcttgaagat
6901 attgtcttga cgctaacact ttttgaggat cgagagatga ttgaggaacg ccttaaaacc
6961 tatgcgcatc tgttcgacga caaagttatg aagcaattga agcgccggag atatacggga
7021 tggggaaggt tgagtcggaa actgataaat ggcatacgcg ataagcagtc cggcaaaaca
7081 attctcgatt tcttgaagtc cgacggattt gcgaatcgaa acttcatgca actgatccat
7141 gacgattcgt tgacctttaa agaggacatt caaaaagccc aagtatccgg acaaggagat
7201 tctttgcacg agcatattgc gaatcttgct ggaagtccgg ctattaaaaa gggcattctc
7261 cagacagtaa aggtcgtgga tgaactcgtt aaagttatgg gccgacacaa acccgagaac
7321 atagtcatcg aaatggcgag agagaaccag accacccaaa agggacagaa aaattcacgc
7381 gagcgaatga agcggattga agaggggatc aaagagcttg gcagccaaat tctgaaagaa
7441 cacccggttg aaaacaccca attgcagaac gagaagctct acctgtacta tcttcaaaat
7501 gggcgtgata tgtacgtcga tcaagaattg gacatcaacc ggctatccga ttacgatgtg
7561 gaccatattg ttcctcagtc gtttctcaaa gacgactcca tagacaataa ggttctaacg
7621 agaagcgaca agaaccgcgg caaatcagac aacgtcccct ccgaggaagt agttaagaaa
7681 atgaagaatt actggcgcca attgctgaac gctaagctca ttacccaaag aaagtttgac
7741 aatttgacga aagccgagcg aggcgggctc tcggaactgg ataaggccgg cttcattaaa
7801 agacaattgg tggaaacgcg acaaatcaca aagcacgtcg cacagattct cgactcccgc
7861 atgaacacta agtatgacga gaatgacaaa ctaatccggg aagtcaaagt gattaccttg
7921 aagtccaagc tggtttccga ttttcgtaaa gattttcagt tttacaaagt tcgcgagatc
7981 aacaactacc atcacgcgca cgacgcttac ttgaacgctg tcgtgggaac ggccttgatt
8041 aaaaaatacc ctaagctgga aagcgagttc gtttacggcg attacaaggt gtatgacgta
8101 cgcaaaatga tcgcgaagtc agagcaggaa attggcaaag ctactgcgaa gtacttcttt
8161 tatagcaaca taatgaattt tttcaaaacg gagattaccc tcgctaacgg cgagattcgc
8221 aagcgccctc ttatcgagac aaacggcgaa acgggggaga ttgtctggga taaaggccgc
8281 gattttgcga ctgtgcggaa agtcttgtcg atgccgcaag tgaatatagt caaaaagacg
8341 gaggtgcaaa caggcggctt ttcgaaagaa tctatccttc ccaaaaggaa ctctgataag
8401 ttgattgcgc gaaaaaagga ttgggaccct aagaagtatg gcggctttga cagcccgacc
8461 gtagcatatt ctgtccttgt ggtggccaaa gtcgaaaagg gcaaatccaa gaaactgaag
8521 agtgtgaaag aattgcttgg gattacgatc atggaaagat catcttttga gaagaatccc
8581 atcgactttc tcgaagctaa aggctacaaa gaagtgaaaa aggacctgat tatcaagttg
8641 cctaagtact ccttgtttga gctggaaaat ggccgtaagc gaatgctcgc ttctgccggc
8701 gaactccaga agggaaacga attggcgctg ccctccaaat atgttaactt tctttacctt
8761 gcctcgcact atgagaagct caagggctcc cccgaggata atgagcaaaa acaactgttt
8821 gtagaacagc ataagcacta cttggacgag atcattgagc aaattagcga gttctccaag
8881 cgagtcatcc ttgccgacgc taatctggac aaagtcctgt ccgcctacaa taagcaccgc
8941 gataagccaa tccgagagca agcggagaat attatccact tgtttacgct caccaatctt
9001 ggagcccctg ccgcctttaa gtactttgac accaccatcg accgcaagag gtacaccagc
9061 accaaagaag tcttggacgc caccctcatt caccaatcaa tcaccggctt gtacgagaca
9121 cgcatcgacc tgtctcaact cggaggcgac aaaaggccgg cggccacgaa aaaggccggc
9181 caggcaaaaa agaaaaagta atctggatcc ctacctcgac tttggctggg acactttcag
9241 tgaggacaag aagctccaga agcgtgctat cgaactcaac cagggacgtg cggcacaaat
9301 gggcatcctt gctctcatgg tgcacgaaca gttgggagtc tctatccttc cttaaaaatt
9361 taattttcat tagttgcagt cactccgctt tggtttcaca gtcaggaata acactagctc
9421 gtcttcactg caggttggct cggaagttgg tgttgacggt gagctggaaa ttggttgtcg
9481 gtcactgcta gcgagaagaa aacggaggac agaaggaagt gaaactcggt tcgttctcga
9541 cagcctcact gtcaatatgc tcattttcaa tccttagcgc ttttaatgtc gaattgacgg
9601 taaattgaat aggatctata atatctacaa ggtactttga cacgccaagt attcattgtt
9661 agtcaacaat attttagagc tttataaggt caaaaaacac cttcaaagtc gaggagaccg
9721 agagagggtc tcagttttag agctagaaat agcaagttaa aataaggcta gtccgttatc
9781 aacttgaaaa agtggcaccg agtcggtgct tttttttaga accgctcacc catgctatcg
9841 tatgtcattt acattgacaa actgctatga atgcgtttgc gataacttca gctcgctgaa
9901 gccctagaca tttcgtttga ctaaattgtt ttccggttgc tttactaccg accggtctgc
9961 gggtagcatg tcttggcttt tgcgagttcg agagaattta tggacaagtg cgtttacagt
10021 ttggaacacg agtgtgagtg agcggaattg gctatctata acattcttat ttcaggaggc
10081 tggtgattag cagaattttc aaagatctcg aatctaaaat cgccatgggc tcgacgaact
10141 tttctgcaag cttatcgata ccgtcgacct cgaggggggg cccggtacca gcttttgttc
10201 cctttagtga gggttaattt cgagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
10261 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
10321 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
10381 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
10441 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
10501 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
10561 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
10621 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
10681 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
10741 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
10801 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
10861 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
10921 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
10981 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
11041 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
11101 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
11161 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
11221 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
11281 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
11341 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
11401 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
11461 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
11521 cagtgctgca atgataccgc gagagccacg ctcaccggct ccagatttat cagcaataaa
11581 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
11641 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
11701 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
11761 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
11821 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
11881 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
11941 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
12001 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
12061 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
12121 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
12181 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
12241 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
12301 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
12361 tccgcgcaca tttccccgaa aagtgccac
//
