| 质粒名称: | pnCasPA-BEC |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 113349 |
| 存储实验室: | Quanjiang Ji |
|
载体骨架基本信息 |
|
| 载体名称: | -- |
| 载体出品公司: | -- |
| 载体抗性: | 庆大霉素 |
| 空载体大小: | 10419 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | CRISPR，合成生物学 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 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: | pnCasPA-BEC was a gift from Quanjiang Ji (Addgene plasmid # 113349 ; http://n2t.net/addgene:113349 ; RRID:Addgene_113349) |
| 参考文献部分: | CRISPR/Cas9-based Genome Editing in Pseudomonas aeruginosa and Cytidine Deaminase-Mediated Base Editing in Pseudomonas Species. Chen W, Zhang Y, Zhang Y, Pi Y, Gu T, Song L, Wang Y, Ji Q. iScience. 2018 Aug 31;6:222-231. doi: 10.1016/j.isci.2018.07.024. Epub 2018 Aug 1. 10.1016/j.isci.2018.07.024 PubMed 30240613 |
买家导航
pnCasPABEC载体基本信息
pnCasPABEC载体图谱质粒图谱和多克隆位点信息
pnCasPABEC载体简介
pnCasPABEC载体序列
LOCUS pnCasPA-BEC 10419 bp ds-DNA circular SYN 07-AUG-2018
DEFINITION A cytidine deaminase-mediated base-editing plasmid in Pseudomonas
species.
ACCESSION .
VERSION .
KEYWORDS pnCasPA-BEC
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10419)
TITLE CRISPR plasmids in Pseudomonas
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 10419)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 7, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..10419
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..30
/label=trc promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
misc_RNA 53..128
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
CDS 323..1009
/codon_start=1
/product="cytidine deaminase (C to U editing enzyme) from
rat"
/label=APOBEC-1
/note="can use ssDNA as a substrate (Komor et al., 2016)"
/translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
LWATGLK"
CDS 1058..5161
/codon_start=1
/product="nickase mutant of the Cas9 endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system"
/label=Cas9(D10A)
/note="generates RNA-guided single strand nicks in DNA due
to the D10A mutation in the RuvC catalytic domain"
/translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
promoter 5283..5311
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"
CDS 5500..6033
/codon_start=1
/gene="aacC1"
/product="gentamycin acetyltransferase"
/label=GmR
/note="confers resistance to gentamycin"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
rep_origin 6331..6682
/label=pRO1600 oriV
/note="broad-host-range origin of replication from
Pseudomonas aeruginosa plasmid pRO1600; requires the
pRO1600 Rep protein for replication (West et al., 1994)"
terminator 7700..7794
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
rep_origin 7882..8470
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8371..8390
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 8561..8580
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
CDS complement(8619..10040)
/codon_start=1
/gene="Bacillus subtilis sacB"
/product="secreted levansucrase that renders bacterial
growth sensitive to sucrose"
/label=SacB
/note="negative selection marker"
/translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
VKDSILEQGQLTVNK"
ORIGIN
1 ttgacaatta atcatccggc tcgtataatg tgtggcgaga ccattggtct cagttttaga
61 gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga
121 gtcggtgctt tttttgagat ctgtccatac ccatggtcta gaatgctcga ggcatcaacg
181 acaagccccg gctatcccga tagccggggc tttgtcttga cggtgtacaa gaacctcttt
241 actattttgt accccgaaaa ttggcagggc gcacgctctg tcatttcgtt tgtgtgtcgt
301 aagacaatca gtggagctat agatgagtag cgaaaccggt ccggttgcag tggatccgac
361 cctgcgccgt cgcattgaac cgcacgagtt tgaagtgttc tttgatccgc gcgagctgcg
421 caaagaaact tgcctgctgt acgagattaa ctggggtggc cgccatagca tctggcgcca
481 taccagccag aacaccaata agcacgtgga agtgaatttt attgaaaaat ttaccaccga
541 gcgctacttc tgccctaata cccgctgcag catcacctgg tttctgagct ggagcccgtg
601 cggcgaatgt agtcgcgcca ttaccgagtt cctgagccgc tatccgcatg tgaccctgtt
661 catctacatc gcccgtctgt accatcacgc cgatccgcgc aatcgccaag gtctgcgtga
721 tctgattagc agcggtgtga ccatccagat catgaccgaa caagagagcg gctattgctg
781 gcgcaacttc gtgaactatt ctccgagcaa cgaagcccac tggccgcgtt atccgcatct
841 gtgggtgcgc ctgtatgtgc tggagctgta ctgcatcatc ctgggcctgc cgccttgcct
901 gaatattctg cgccgtaaac agccgcaact gacattcttc accatcgcac tgcagagctg
961 ccattatcag cgcctgccgc cgcacatttt atgggccacc ggtctgaaaa gcggtagtga
1021 aactccgggc acaagcgaaa gcgcaacccc ggaaagtgat aagaaatact caataggctt
1081 agctatcggc acaaatagcg tcggatgggc ggtgatcact gatgaatata aggttccgtc
1141 taaaaagttc aaggttctgg gaaatacaga ccgccacagt atcaaaaaaa atcttatagg
1201 ggctctttta tttgacagtg gagagacagc ggaagcgact cgtctcaaac ggacagctcg
1261 tagaaggtat acacgtcgga agaatcgtat ttgttatcta caggagattt tttcaaatga
1321 gatggcgaaa gtagatgata gtttctttca tcgacttgaa gagtcttttt tggtggaaga
1381 agacaagaag catgaacgtc atcctatttt tggaaatata gtagatgaag ttgcttatca
1441 tgagaaatat ccaactatct atcatctgcg aaaaaaattg gtagattcta ctgataaagc
1501 ggatttgcgc ttaatctatt tggccttagc gcatatgatt aagtttcgtg gtcatttttt
1561 gattgaggga gatttaaatc ctgataatag tgatgtggac aaactattta tccagttggt
1621 acaaacctac aatcaattat ttgaagaaaa ccctattaac gcaagtggag tagatgctaa
1681 agcgattctt tctgcacgat tgagtaaatc aagacgatta gaaaatctca ttgctcagct
1741 ccccggtgag aagaaaaatg gcttatttgg gaatctcatt gctttgtcat tgggtttgac
1801 ccctaatttt aaatcaaatt ttgatttggc agaagatgct aaattacagc tttcaaaaga
1861 tacttacgat gatgatttag ataatttatt ggcgcaaatt ggagatcaat atgctgattt
1921 gtttttggca gctaagaatt tatcagatgc tattttactt tcagatatcc taagagtaaa
1981 tactgaaata actaaggctc ccctatcagc ttcaatgatt aaacgctacg atgaacatca
2041 tcaagacttg actcttttaa aagctttagt tcgacaacaa cttccagaaa agtataaaga
2101 aatctttttt gatcaatcaa aaaacggata tgcaggttat attgatgggg gagctagcca
2161 agaagaattt tataaattta tcaaaccaat tttagaaaaa atggatggta ctgaggaatt
2221 attggtgaaa ctaaatcgtg aagatttgct gcgcaagcaa cggacctttg acaacggctc
2281 tattccccat caaattcact tgggtgagct gcatgctatt ttgagaagac aagaagactt
2341 ttatccattt ttaaaagaca atcgtgagaa gattgaaaaa atcttgactt ttcgaattcc
2401 ttattatgtt ggtccattgg cgcgtggcaa tagtcgtttt gcatggatga ctcggaagtc
2461 tgaagaaaca attaccccat ggaattttga agaagttgtc gataaaggtg cttcagctca
2521 atcatttatt gaacgcatga caaactttga taaaaatctt ccaaatgaaa aagtactacc
2581 aaaacatagt ttgctttatg agtattttac ggtttataac gaattgacaa aggtcaaata
2641 tgttactgaa ggaatgcgaa aaccagcatt tctttcaggt gaacagaaga aagccattgt
2701 tgatttactc ttcaaaacaa atcgaaaagt aaccgttaag caattaaaag aagattattt
2761 caaaaaaata gaatgttttg atagtgttga aatttcagga gttgaagata gatttaatgc
2821 ttcattaggt acctaccatg atttgctaaa aattattaaa gataaagatt ttttggataa
2881 tgaagaaaat gaagatatct tagaggatat tgttttaaca ttgaccttat ttgaagatag
2941 ggagatgatt gaggaaagac ttaaaacata tgctcacctc tttgatgata aggtgatgaa
3001 acagcttaaa cgtcgccgtt atactggttg gggacgtttg tctcgaaaat tgattaatgg
3061 tattagggat aagcaatctg gcaaaacaat attagatttt ttgaaatcag atggttttgc
3121 caatcgcaat tttatgcagc tgatccatga tgatagtttg acatttaaag aagacattca
3181 aaaagcacaa gtgtctggac aaggcgatag tttacatgaa catattgcaa atttagctgg
3241 tagccctgct attaaaaaag gtattttaca gactgtaaaa gttgttgatg aattggtcaa
3301 agtaatgggg cggcataagc cagaaaatat cgttattgaa atggcacgtg aaaatcagac
3361 aactcaaaag ggccagaaaa attcgcgaga gcgtatgaaa cgaatcgaag aaggtatcaa
3421 agaattagga agtcagattc ttaaagagca tcctgttgaa aatactcaat tgcaaaatga
3481 aaagctctat ctctattatc tccaaaatgg aagagacatg tatgtggacc aagaattaga
3541 tattaatcgt ttaagtgatt atgatgtcga tcacattgtt ccacaaagtt tccttaaaga
3601 cgattcaata gacaataagg tcttaacgcg ttctgataaa aatcgtggta aatcggataa
3661 cgttccaagt gaagaagtag tcaaaaagat gaaaaactat tggagacaac ttctaaacgc
3721 caagttaatc actcaacgta agtttgataa tttaacgaaa gctgaacgtg gaggtttgag
3781 tgaacttgat aaagctggtt ttatcaaacg ccaattggtt gaaactcgcc aaatcactaa
3841 gcatgtggca caaattttgg atagtcgcat gaatactaaa tacgatgaaa atgataaact
3901 tattcgagag gttaaagtga ttaccttaaa atctaaatta gtttctgact tccgaaaaga
3961 tttccaattc tataaagtac gtgagattaa caattaccat catgcccatg atgcgtatct
4021 aaatgccgtc gttggaactg ctttgattaa gaaatatcca aaacttgaat cggagtttgt
4081 ctatggtgat tataaagttt atgatgttcg taaaatgatt gctaagtctg agcaagaaat
4141 aggcaaagca accgcaaaat atttctttta ctctaatatc atgaacttct tcaaaacaga
4201 aattacactt gcaaatggag agattcgcaa acgccctcta atcgaaacta atggggaaac
4261 tggagaaatt gtctgggata aagggcgaga ttttgccaca gtgcgcaaag tattgtccat
4321 gccccaagtc aatattgtca agaaaacaga agtacagaca ggcggattct ccaaggagtc
4381 aattttacca aaaagaaatt cggacaagct tattgctcgt aaaaaagact gggatccaaa
4441 aaaatatggt ggttttgata gtccaacggt agcttattca gtcctagtgg ttgctaaggt
4501 ggaaaaaggg aaatcgaaga agttaaaatc cgttaaagag ttactaggga tcacaattat
4561 ggaaagaagt tcctttgaaa aaaatccgat tgacttttta gaagctaaag gatataagga
4621 agttaaaaaa gacttaatca ttaaactacc taaatatagt ctttttgagt tagaaaacgg
4681 tcgtaaacgg atgctggcta gtgccggaga attacaaaaa ggaaatgagc tggctctgcc
4741 aagcaaatat gtgaattttt tatatttagc tagtcattat gaaaagttga agggtagtcc
4801 agaagataac gaacaaaaac aattgtttgt ggagcagcat aagcattatt tagatgagat
4861 tattgagcaa atcagtgaat tttctaagcg tgttatttta gcagatgcca atttagataa
4921 agttcttagt gcatataaca aacatagaga caaaccaata cgtgaacaag cagaaaatat
4981 tattcattta tttacgttga cgaatcttgg agctcccgct gcttttaaat attttgatac
5041 aacaattgat cgtaaacgat atacgtctac aaaagaagtt ttagatgcca ctcttatcca
5101 tcaatccatc actggtcttt atgaaacacg cattgatttg agtcagctag gaggtgactg
5161 atggctggtt ggcgtactgt tgtggtaaat acccatgggt atggacagtt ttcccgaata
5221 ggaacttcgg aataggaact tcaagatccc ctgattccct ttgtcaacag caatggatcg
5281 aattgacata agcctgttcg gttcgtaaac tgtaatgcaa gtagcgtatg cgctcacgca
5341 actggtccag aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg
5401 cttgttatga ctgttttttt gtacagtcta tgcctcgggc atccaagcag caagcgcgtt
5461 acgccgtggg tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagcaacgat
5521 gttacgcagc agggcagtcg ccctaaaaca aagttaggtg gctcaagtat gggcatcatt
5581 cgcacatgta ggctcggccc tgaccaagtc aaatccatgc gggctgctct tgatcttttc
5641 ggtcgtgagt tcggagacgt agccacctac tcccaacatc agccggactc cgattacctc
5701 gggaacttgc tccgtagtaa gacattcatc gcgcttgctg ccttcgacca agaagcggtt
5761 gttggcgctc tcgcggctta cgttctgccc aagtttgagc agccgcgtag tgagatctat
5821 atctatgatc tcgcagtctc cggcgagcac cggaggcagg gcattgccac cgcgctcatc
5881 aatctcctca agcatgaggc caacgcgctt ggtgcttatg tgatctacgt gcaagcagat
5941 tacggtgacg atcccgcagt ggctctctat acaaagttgg gcatacggga agaagtgatg
6001 cactttgata tcgacccaag taccgccacc taacaattcg ttcaagccga gatcagcttc
6061 ccggccgcgg agttgttcgg taaattgtca caacgccgcg gccaattcga gagttttcgt
6121 tccactgagc gtcagacccc aattacacgc cactggctgt gcttgctggg gtgacggtgg
6181 caacggtggc ggccttgctg ggctatcgcg ttggaaagaa acgagggaaa ggggactgat
6241 aaaccggtct tagcccctcc ccttggtgtc caaccgctct gtaggcctct caggcgccgc
6301 tggtgccgct ggttggacgc caagggtgaa tccgcctcga taccctgatt actcgcttcc
6361 tgcgccctct caggcggcga taggggactg gtaaaacggg gattgcccag acgcctcccc
6421 cgccccttca ggggcacaaa tgcggcccca acggggccac gtagtggtgc gttttttgcg
6481 tttccaccct tttcttcctt ttccctttta aaccttttag gacgtctaca ggccacgtaa
6541 tccgtggcct gtagagttta aaaagggacg gatttgttgc cattaaggga cggatttgtt
6601 gttaagaagg gacggatttg ttgttgtaaa gggacggatt tgttgtattg tgggacgcag
6661 atacagtgtc cccttataca caaggaatgt cgaacgtggc ctcaccccca atggtttaca
6721 aaagcaatgc cctggtcgag gccgcgtatc gcctcagtgt tcaggaacag cggatcgttc
6781 tggcctgtat tagccaggtg aagaggagcg agcctgtcac cgatgaagtg atgtattcag
6841 tgacggcgga ggacatagcg acgatggcgg gtgtccctat cgaatcttcc tacaaccagc
6901 tcaaagaagc ggccctgcgc ctgaaacggc gggaagtccg gttaacccaa gagcccaatg
6961 gcaaggggaa aagaccgagt gtgatgatta ccggctgggt gcaaacaatc atctaccggg
7021 agggtgaggg ccgtgtagaa ctcaggttca ccaaagacat gctgccgtac ctgacggaac
7081 tcaccaaaca gttcaccaaa tacgccttgg ctgacgtggc caagatggac agcacccacg
7141 cgatcaggct ttacgagctg ctcatgcaat gggacagcat cggccagcgc gaaatagaaa
7201 ttgaccagct gcgaaagtgg tttcaactgg aaggccggta tccctcgatc aaggacttca
7261 agttgcgagt gcttgatcca gccgtgacgc agatcaacga gcacagcccg ctacaggtgg
7321 agtgggcgca gcgaaagacc gggcgcaagg tcacacatct gttgttcagt tttggaccga
7381 agaagcccgc caaggcggtg ggtaaggccc cagcgaagcg caaggccggg aagatttcag
7441 atgctgagat cgcgaaacag gctcgccctg gtgagacatg ggaagcggcc cgcgctcgac
7501 taacccagat gccgcttgcg caaactatta actggcgaac tacttactct agcttcccgg
7561 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
7621 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg cgggactctg
7681 gggttcgaga gctcgcttgg actcctgttg atagatccag taatgacctc agaactccat
7741 ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtga gaatccaagc
7801 actagtcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
7861 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
7921 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
7981 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
8041 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
8101 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
8161 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
8221 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
8281 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
8341 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
8401 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
8461 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
8521 tgctcacatg ttctttcctg cgttgtcgac agagcccggg cggcatcaga gcagattgta
8581 ctgagagtgc accataatcg gcattttctt ttgcgttttt atttgttaac tgttaattgt
8641 ccttgttcaa ggatgctgtc tttgacaaca gatgttttct tgcctttgat gttcagcagg
8701 aagctaggcg caaacgttga ttgtttgtct gcgtagaatc ctctgtttgt catatagctt
8761 gtaatcacga cattgtttcc tttcgcttga ggtacagcga agtgtgagta agtaaaggtt
8821 acatcgttag gatcaagatc catttttaac acaaggccag ttttgttcag cggcttgtat
8881 gggccagtta aagaattaga aacataacca agcatgtaaa tatcgttaga cgtaatgccg
8941 tcaatcgtca tttttgatcc gcgggagtca gtgaacagat accatttgcc gttcatttta
9001 aagacgttcg cgcgttcaat ttcatctgtt actgtgttag atgcaatcag cggtttcatc
9061 acttttttca gtgtgtaatc atcgtttagc tcaatcatac cgagagcgcc gtttgctaac
9121 tcagccgtgc gttttttatc gctttgcaga agtttttgac tttcttgacg gaagaatgat
9181 gtgcttttgc catagtatgc tttgttaaat aaagattctt cgccttggta gccatcttca
9241 gttccagtgt ttgcttcaaa tactaagtat ttgtggcctt tatcttctac gtagtgagga
9301 tctctcagcg tatggttgtc gcctgagctg tagttgcctt catcgatgaa ctgctgtaca
9361 ttttgatacg tttttccgtc accgtcaaag attgatttat aatcctctac accgttgatg
9421 ttcaaagagc tgtctgatgc tgatacgtta acttgtgcag ttgtcagtgt ttgtttgccg
9481 taatgtttac cggagaaatc agtgtagaat aaacggattt ttccgtcaga tgtaaatgtg
9541 gctgaacctg accattcttg tgtttggtct tttaggatag aatcatttgc atcgaatttg
9601 tcgctgtctt taaagacgcg gccagcgttt ttccagctgt caatagaagt ttcgccgact
9661 ttttgataga acatgtaaat cgatgtgtca tccgcatttt taggatctcc ggctaatgca
9721 aagacgatgt ggtagccgtg atagtttgcg acagtgccgt cagcgttttg taatggccag
9781 ctgtcccaaa cgtccaggcc ttttgcagaa gagatatttt taattgtgga cgaatcgaac
9841 tcaggaactt gatatttttc atttttttgc tgttcaggga tttgcagcat atcatggcgt
9901 gtaatatggg aaatgccgta tgtttcctta tatggctttt ggttcgtttc tttcgcaaac
9961 gcttgagttg cgcctcctgc cagcagtgcg gtagtaaagg ttaatactgt tgcttgtttt
10021 gcaaactttt tgatgttcat cgttcatgtc tcctttttta tgtactgtgt tagcggtctg
10081 cttcttccag ccctcctgtt tgaagatggc aagttagtta cgcacaataa aaaaagacct
10141 aaaatatgta aggggtgacg ccaaagtata cactttgccc tttacacatt ttaggtcttg
10201 cctgctttat cagtaacaaa cccgcgcgat ttacttttcg acctcattct attagactct
10261 cgtttggatt gcaactggtc tattttcctc ttttgtttga tagaaaatca taaaaggatt
10321 tgcagactac gggcctaaag aactaaaaaa tctatctgtt tcttttcatt ctctgtattt
10381 tttatagttt ctgttgcatg ggcataaagt tgccttaag
//
