| 质粒名称: | pET302-6His-dCas9-Halo |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 72269 |
| 存储实验室: | Timothée Lionnet |
|
载体骨架基本信息 |
|
| 载体名称: | pET302 |
| 载体出品公司: | -- |
| 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: | -- |
| 筛选标记: | -- |
| 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: | His6-dCas9-Halo |
| 物种来源: | 合成 |
| 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: | pET302-6His-dCas9-Halo was a gift from Timothée Lionnet (Addgene plasmid # 72269 ; http://n2t.net/addgene:72269 ; RRID:Addgene_72269) |
| 参考文献部分: | CASFISH: CRISPR/Cas9-mediated in situ labeling of genomic loci in fixed cells. Deng W, Shi X, Tjian R, Lionnet T, Singer RH. Proc Natl Acad Sci U S A. 2015 Sep 22;112(38):11870-5. doi: 10.1073/pnas.1515692112. Epub 2015 Aug 31. 10.1073/pnas.1515692112 PubMed 26324940 |
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 pET302-6His-dCas9-Halo 10715 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Encodes a Halo-labeled fusion of nuclease deficient Cas9.
ACCESSION .
VERSION .
KEYWORDS pET302-6His-dCas9-Halo
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10715)
AUTHORS Deng W, Shi X, Tjian R, Lionnet T, Singer RH
TITLE CASFISH: CRISPR/Cas9-mediated in situ labeling of genomic loci in
fixed cells.
JOURNAL Proc Natl Acad Sci U S A. 2015 Sep 22;112(38):11870-5. doi:
10.1073/pnas.1515692112. Epub 2015 Aug 31.
PUBMED 26324940
REFERENCE 2 (bases 1 to 10715)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..10715
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 62..81
/label=T7
/note="T7 promoter, forward primer"
promoter 62..80
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 81..105
/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)."
CDS 153..170
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 174..4274
/codon_start=1
/product="catalytically dead mutant of the Cas9
endonuclease from the Streptococcus pyogenes Type II
CRISPR/Cas system"
/label=dCas9
/note="RNA-guided DNA-binding protein that lacks
endonuclease activity due to the D10A mutation in the RuvC
catalytic domain and the H840A mutation in the HNH
catalytic domain"
/translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 4290..5174
/codon_start=1
/product="modified bacterial dehalogenase that forms
covalent bonds with chloroalkane derivatives"
/label=HaloTag(R)
/translation="AEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
GSEIARWLSTLEIS"
primer_bind complement(5269..5287)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 5283..5330
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind complement(5568..5586)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5654..5758
/gene="bla"
/label=AmpR promoter
CDS 5759..6619
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(5977..5996)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6790..7378
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7279..7298
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7532..7549
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 7564..7703
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(7629..7648)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 7789..7811
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(7805..7996)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(9308..10390)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind 10352..10371
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(10391..10468)
/gene="lacI"
/label=lacI promoter
primer_bind 10674..10693
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
ORIGIN
1 gccggtgatg ccggccacga tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa
61 ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctct agaaataatt
121 ttgtttaaac tttaagaagg agatatacta tgcatcatca tcatcatcac gtggataaga
181 aatactcaat aggcttagct atcggcacaa atagcgtcgg atgggcggtg atcactgatg
241 aatataaggt tccgtctaaa aagttcaagg ttctgggaaa tacagaccgc cacagtatca
301 aaaaaaatct tataggggct cttttatttg acagtggaga gacagcggaa gcgactcgtc
361 tcaaacggac agctcgtaga aggtatacac gtcggaagaa tcgtatttgt tatctacagg
421 agattttttc aaatgagatg gcgaaagtag atgatagttt ctttcatcga cttgaagagt
481 cttttttggt ggaagaagac aagaagcatg aacgtcatcc tatttttgga aatatagtag
541 atgaagttgc ttatcatgag aaatatccaa ctatctatca tctgcgaaaa aaattggtag
601 attctactga taaagcggat ttgcgcttaa tctatttggc cttagcgcat atgattaagt
661 ttcgtggtca ttttttgatt gagggagatt taaatcctga taatagtgat gtggacaaac
721 tatttatcca gttggtacaa acctacaatc aattatttga agaaaaccct attaacgcaa
781 gtggagtaga tgctaaagcg attctttctg cacgattgag taaatcaaga cgattagaaa
841 atctcattgc tcagctcccc ggtgagaaga aaaatggctt atttgggaat ctcattgctt
901 tgtcattggg tttgacccct aattttaaat caaattttga tttggcagaa gatgctaaat
961 tacagctttc aaaagatact tacgatgatg atttagataa tttattggcg caaattggag
1021 atcaatatgc tgatttgttt ttggcagcta agaatttatc agatgctatt ttactttcag
1081 atatcctaag agtaaatact gaaataacta aggctcccct atcagcttca atgattaaac
1141 gctacgatga acatcatcaa gacttgactc ttttaaaagc tttagttcga caacaacttc
1201 cagaaaagta taaagaaatc ttttttgatc aatcaaaaaa cggatatgca ggttatattg
1261 atgggggagc tagccaagaa gaattttata aatttatcaa accaatttta gaaaaaatgg
1321 atggtactga ggaattattg gtgaaactaa atcgtgaaga tttgctgcgc aagcaacgga
1381 cctttgacaa cggctctatt ccccatcaaa ttcacttggg tgagctgcat gctattttga
1441 gaagacaaga agacttttat ccatttttaa aagacaatcg tgagaagatt gaaaaaatct
1501 tgacttttcg aattccttat tatgttggtc cattggcgcg tggcaatagt cgttttgcat
1561 ggatgactcg gaagtctgaa gaaacaatta ccccatggaa ttttgaagaa gttgtcgata
1621 aaggtgcttc agctcaatca tttattgaac gcatgacaaa ctttgataaa aatcttccaa
1681 atgaaaaagt actaccaaaa catagtttgc tttatgagta ttttacggtt tataacgaat
1741 tgacaaaggt caaatatgtt actgaaggaa tgcgaaaacc agcatttctt tcaggtgaac
1801 agaagaaagc cattgttgat ttactcttca aaacaaatcg aaaagtaacc gttaagcaat
1861 taaaagaaga ttatttcaaa aaaatagaat gttttgatag tgttgaaatt tcaggagttg
1921 aagatagatt taatgcttca ttaggtacct accatgattt gctaaaaatt attaaagata
1981 aagatttttt ggataatgaa gaaaatgaag atatcttaga ggatattgtt ttaacattga
2041 ccttatttga agatagggag atgattgagg aaagacttaa aacatatgct cacctctttg
2101 atgataaggt gatgaaacag cttaaacgtc gccgttatac tggttgggga cgtttgtctc
2161 gaaaattgat taatggtatt agggataagc aatctggcaa aacaatatta gattttttga
2221 aatcagatgg ttttgccaat cgcaatttta tgcagctgat ccatgatgat agtttgacat
2281 ttaaagaaga cattcaaaaa gcacaagtgt ctggacaagg cgatagttta catgaacata
2341 ttgcaaattt agctggtagc cctgctatta aaaaaggtat tttacagact gtaaaagttg
2401 ttgatgaatt ggtcaaagta atggggcggc ataagccaga aaatatcgtt attgaaatgg
2461 cacgtgaaaa tcagacaact caaaagggcc agaaaaattc gcgagagcgt atgaaacgaa
2521 tcgaagaagg tatcaaagaa ttaggaagtc agattcttaa agagcatcct gttgaaaata
2581 ctcaattgca aaatgaaaag ctctatctct attatctcca aaatggaaga gacatgtatg
2641 tggaccaaga attagatatt aatcgtttaa gtgattatga tgtcgatgcc attgttccac
2701 aaagtttcct taaagacgat tcaatagaca ataaggtctt aacgcgttct gataaaaatc
2761 gtggtaaatc ggataacgtt ccaagtgaag aagtagtcaa aaagatgaaa aactattgga
2821 gacaacttct aaacgccaag ttaatcactc aacgtaagtt tgataattta acgaaagctg
2881 aacgtggagg tttgagtgaa cttgataaag ctggttttat caaacgccaa ttggttgaaa
2941 ctcgccaaat cactaagcat gtggcacaaa ttttggatag tcgcatgaat actaaatacg
3001 atgaaaatga taaacttatt cgagaggtta aagtgattac cttaaaatct aaattagttt
3061 ctgacttccg aaaagatttc caattctata aagtacgtga gattaacaat taccatcatg
3121 cccatgatgc gtatctaaat gccgtcgttg gaactgcttt gattaagaaa tatccaaaac
3181 ttgaatcgga gtttgtctat ggtgattata aagtttatga tgttcgtaaa atgattgcta
3241 agtctgagca agaaataggc aaagcaaccg caaaatattt cttttactct aatatcatga
3301 acttcttcaa aacagaaatt acacttgcaa atggagagat tcgcaaacgc cctctaatcg
3361 aaactaatgg ggaaactgga gaaattgtct gggataaagg gcgagatttt gccacagtgc
3421 gcaaagtatt gtccatgccc caagtcaata ttgtcaagaa aacagaagta cagacaggcg
3481 gattctccaa ggagtcaatt ttaccaaaaa gaaattcgga caagcttatt gctcgtaaaa
3541 aagactggga tccaaaaaaa tatggtggtt ttgatagtcc aacggtagct tattcagtcc
3601 tagtggttgc taaggtggaa aaagggaaat cgaagaagtt aaaatccgtt aaagagttac
3661 tagggatcac aattatggaa agaagttcct ttgaaaaaaa tccgattgac tttttagaag
3721 ctaaaggata taaggaagtt aaaaaagact taatcattaa actacctaaa tatagtcttt
3781 ttgagttaga aaacggtcgt aaacggatgc tggctagtgc cggagaatta caaaaaggaa
3841 atgagctggc tctgccaagc aaatatgtga attttttata tttagctagt cattatgaaa
3901 agttgaaggg tagtccagaa gataacgaac aaaaacaatt gtttgtggag cagcataagc
3961 attatttaga tgagattatt gagcaaatca gtgaattttc taagcgtgtt attttagcag
4021 atgccaattt agataaagtt cttagtgcat ataacaaaca tagagacaaa ccaatacgtg
4081 aacaagcaga aaatattatt catttattta cgttgacgaa tcttggagct cccgctgctt
4141 ttaaatattt tgatacaaca attgatcgta aacgatatac gtctacaaaa gaagttttag
4201 atgccactct tatccatcaa tccatcactg gtctttatga aacacgcatt gatttgagtc
4261 agctaggagg tgacggtggc tccagatctg cagaaatcgg tactggcttt ccattcgacc
4321 cccattatgt ggaagtcctg ggcgagcgca tgcactacgt cgatgttggt ccgcgcgatg
4381 gcacccctgt gctgttcctg cacggtaacc cgacctcctc ctacgtgtgg cgcaacatca
4441 tcccgcatgt tgcaccgacc catcgctgca ttgctccaga cctgatcggt atgggcaaat
4501 ccgacaaacc agacctgggt tatttcttcg acgaccacgt ccgcttcatg gatgccttca
4561 tcgaagccct gggtctggaa gaggtcgtcc tggtcattca cgactggggc tccgctctgg
4621 gtttccactg ggccaagcgc aatccagagc gcgtcaaagg tattgcattt atggagttca
4681 tccgccctat cccgacctgg gacgaatggc cagaatttgc ccgcgagacc ttccaggcct
4741 tccgcaccac cgacgtcggc cgcaagctga tcatcgatca gaacgttttt atcgagggta
4801 cgctgccgat gggtgtcgtc cgcccgctga ctgaagtcga gatggaccat taccgcgagc
4861 cgttcctgaa tcctgttgac cgcgagccac tgtggcgctt cccaaacgag ctgccaatcg
4921 ccggtgagcc agcgaacatc gtcgcgctgg tcgaagaata catggactgg ctgcaccagt
4981 cccctgtccc gaagctgctg ttctggggca ccccaggcgt tctgatccca ccggccgaag
5041 ccgctcgcct ggccaaaagc ctgcctaact gcaaggctgt ggacatcggc ccgggtctga
5101 atctgctgca agaagacaac ccggacctga tcggcagcga gatcgcgcgc tggctgtcta
5161 ctctggagat ttccggttaa tagctcgaga tcgatgatat tcgagcctag gtataatcgg
5221 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
5281 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
5341 gaactatatc cggatatccc gcaagaggcc cggcagtacc ggcataacca agcctatgcc
5401 tacagcatcc agggtgacgg tgccgaggat gacgatgagc gcattgttag atttcataca
5461 cggtgcctga ctgcgttagc aatttaactg tgataaacta ccgcattaaa gctagcttat
5521 cgatgataag ctgtcaaaca tgagaattaa ttcttgaaga cgaaagggcc tcgtgatacg
5581 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
5641 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
5701 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
5761 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
5821 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
5881 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
5941 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
6001 tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
6061 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
6121 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
6181 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
6241 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
6301 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
6361 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
6421 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
6481 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
6541 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
6601 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
6661 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
6721 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
6781 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
6841 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
6901 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
6961 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
7021 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
7081 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
7141 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
7201 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
7261 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
7321 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
7381 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
7441 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
7501 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
7561 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc
7621 actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc
7681 tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac
7741 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
7801 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat
7861 cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga
7921 gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt
7981 tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga
8041 taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt
8101 tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa
8161 tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc
8221 agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt
8281 ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg
8341 ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag
8401 taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc
8461 gtggccagga cccaacgctg cccgagatgc gccgcgtgcg gctgctggag atggcggacg
8521 cgatggatat gttctgccaa gggttggttt gcgcattcac agttctccgc aagaattgat
8581 tggctccaat tcttggagtg gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt
8641 cgaggtggcc cggctccatg caccgcgacg caacgcgggg aggcagacaa ggtatagggc
8701 ggcgcctaca atccatgcca acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt
8761 gacgatcagc ggtccaatga tcgaagttag gctggtaaga gccgcgagcg atccttgaag
8821 ctgtccctga tggtcgtcat ctacctgcct ggacagcatg gcctgcaacg cgggcatccc
8881 gatgccgccg gaagcgagaa gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa
8941 cgccagcaag acgtagccca gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc
9001 gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc
9061 gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa
9121 aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat
9181 aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
9241 tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc
9301 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
9361 cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca
9421 ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca
9481 agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg
9541 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa
9601 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
9661 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
9721 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
9781 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca
9841 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt
9901 catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg
9961 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa
10021 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga
10081 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt
10141 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa
10201 tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct
10261 ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca
10321 ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta
10381 ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc
10441 gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc
10501 tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat
10561 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
10621 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
10681 tcggcgatat aggcgccagc aaccgcacct gtggc
//
