| 质粒名称: | pCSN067 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 101748 |
| 存储实验室: | Rene Verwaal |
|
载体骨架基本信息 |
|
| 载体名称: | pRS414 |
| 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: | 11322 bp |
| 载体修饰: | -- |
| 载体类型: | 酵母表达,CRISPR |
| 筛选标记: | TRP1 ; KanMX |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Cpf1 from Lachnospiraceae bacterium ND2006 (LbCpf1) codon optimized for expression in S. cerevisiae.LbCpf1 |
| 物种来源: | Lachnospiraceae bacterium ND2006 |
| 插入大小: | 3720 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | Heterologous promoter from K. lactis (KLLA0F20031g). |
| 标签/融合蛋白: | SV40 NLS（插接件上的C端子） |
| 克隆方法: | Gibson克隆 |
| 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: | pCSN067 was a gift from Rene Verwaal (Addgene plasmid # 101748 ; http://n2t.net/addgene:101748 ; RRID:Addgene_101748) |
| 参考文献部分: | CRISPR/Cpf1 enables fast and simple genome editing of Saccharomyces cerevisiae. Verwaal R, Buiting-Wiessenhaan N, Dalhuijsen S, Roubos JA. Yeast. 2017 Sep 8. doi: 10.1002/yea.3278. 10.1002/yea.3278 PubMed 28886218 |
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 pCSN067 11326 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Single copy yeast plasmid expressing Cpf1 from Lachnospiraceae
bacterium ND2006 (LbCpf1), codon optimized for expression in
Saccharomyces cerevisiae..
ACCESSION .
VERSION .
KEYWORDS pCSN067
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11326)
AUTHORS Verwaal R, Buiting-Wiessenhaan N, Dalhuijsen S, Roubos JA
TITLE CRISPR/Cpf1 enables fast and simple genome editing of Saccharomyces
cerevisiae.
JOURNAL Yeast. 2017 Sep 8. doi: 10.1002/yea.3278.
PUBMED 28886218
REFERENCE 2 (bases 1 to 11326)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..11326
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 965..4648
/codon_start=1
/product="CRISPR-associated protein from Lachnospiraceae
bacterium ND2006 (Zetsche et al., 2015)"
/label=LbCpf1
/translation="MSKLEKFTNCYSLSKTLRFKAIPVGKTQENIDNKRLLVEDEKRAE
DYKGVKKLLDRYYLSFINDVLHSIKLKNLNNYISLFRKKTRTEKENKELENLEINLRKE
IAKAFKGNEGYKSLFKKDIIETILPEFLDDKDEIALVNSFNGFTTAFTGFFDNRENMFS
EEAKSTSIAFRCINENLTRYISNMDIFEKVDAIFDKHEVQEIKEKILNSDYDVEDFFEG
EFFNFVLTQEGIDVYNAIIGGFVTESGEKIKGLNEYINLYNQKTKQKLPKFKPLYKQVL
SDRESLSFYGEGYTSDEEVLEVFRNTLNKNSEIFSSIKKLEKLFKNFDEYSSAGIFVKN
GPAISTISKDIFGEWNVIRDKWNAEYDDIHLKKKAVVTEKYEDDRRKSFKKIGSFSLEQ
LQEYADADLSVVEKLKEIIIQKVDEIYKVYGSSEKLFDADFVLEKSLKKNDAVVAIMKD
LLDSVKSFENYIKAFFGEGKETNRDESFYGDFVLAYDILLKVDHIYDAIRNYVTQKPYS
KDKFKLYFQNPQFMGGWDKDKETDYRATILRYGSKYYLAIMDKKYAKCLQKIDKDDVNG
NYEKINYKLLPGPNKMLPKVFFSKKWMAYYNPSEDIQKIYKNGTFKKGDMFNLNDCHKL
IDFFKDSISRYPKWSNAYDFNFSETEKYKDIAGFYREVEEQGYKVSFESASKKEVDKLV
EEGKLYMFQIYNKDFSDKSHGTPNLHTMYFKLLFDENNHGQIRLSGGAELFMRRASLKK
EELVVHPANSPIANKNPDNPKKTTTLSYDVYKDKRFSEDQYELHIPIAINKCPKNIFKI
NTEVRVLLKHDDNPYVIGIDRGERNLLYIVVVDGKGNIVEQYSLNEIINNFNGIRIKTD
YHSLLDKKEKERFEARQNWTSIENIKELKAGYISQVVHKICELVEKYDAVIALEDLNSG
FKNSRVKVEKQVYQKFEKMLIDKLNYMVDKKSNPCATGGALKGYQITNKFESFKSMSTQ
NGFIFYIPAWLTSKIDPSTGFVNLLKTKYTSIADSKKFISSFDRIMYVPEEDLFEFALD
YKNFSRTDADYIKKWKLYSYGNRIRIFRNPKKNNVFDWEEVCLTSAYKELFNKYGINYQ
QGDIRALLCEQSDKAFYSSFMALMSLMLQMRNSITGRTDVDFLISPVKNSDGIFYDSRN
YEAQENAILPKNADANGAYNIARKVLWAIGQFKKAEDEKLDKVKIAISNKEWLEYAQTS
VKH"
CDS 4661..4681
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
protein_bind complement(5030..5063)
/label=lox66
/bound_moiety="Cre recombinase"
/note="loxP mutant. Cre-mediated recombination occurs in
the 8-bp core sequence (GCATACAT)."
gene 5109..6465
/label=kanMX
/note="yeast selectable marker conferring kanamycin
resistance (Wach et al., 1994)"
promoter 5109..5452
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
CDS 5453..6262
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind complement(5520..5539)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
terminator 6268..6465
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
protein_bind complement(6513..6546)
/label=lox71
/bound_moiety="Cre recombinase"
/note="loxP mutant. Cre-mediated recombination occurs in
the 8-bp core sequence (GCATACAT)."
primer_bind complement(6614..6634)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(6614..6632)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(6653..6669)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6653..6669)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6666..6688)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6677..6693
/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(6701..6731)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6746..6767
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6884..6901)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(7055..7643)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7135..7154)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7814..8674)
/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 8437..8456
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8675..8779)
/gene="bla"
/label=AmpR promoter
misc_feature 8816..9319
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
primer_bind 9360..9378
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(9416..9438)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 9538..9557
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 9578..9858
/gene="S. cerevisiae TRP1"
/label=TRP1 promoter
CDS 9859..10533
/codon_start=1
/gene="S. cerevisiae TRP1"
/product="phosphoribosylanthranilate isomerase, required
for tryptophan biosynthesis"
/label=TRP1
/note="yeast auxotrophic marker"
/translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
KK"
rep_origin complement(10636..11091)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(10773..10794)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 10985..11004
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 11217..11239
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 11231..11248
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 11232..11248
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 11258..11277
/label=T7
/note="T7 promoter, forward primer"
promoter 11258..11276
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 ggcagaggag taggtagagc aacgaatcct actatttatc caaattagtc taggaactct
61 ttttctagat tttttagatt tgagggcaag cgctgttaac gactcagaaa tgtaagcact
121 acggagtaga acgagaaatc cgccataggt ggaaatccta gcaaaatctt gcttacccta
181 gctagcctca ggtaagctag ccttagcctg tcaaattttt ttcaaaattt ggtaagtttc
241 tactagcaaa gcaaacacgg ttcaacaaac cgaaaactcc actcattata cgtggaaacc
301 gaaacaaaaa aacaaaaacc aaaatactcg ccaatgagaa agttgctgcg tttctacttt
361 cgaggaagag gaactgagag gattgactac gaaaggggca aaaacgagtc gtattctccc
421 attattgtct gctaccacgc ggtctagtag aataagcaac cagtcaacgc taagacaggt
481 aatcaaaata ccagtctgct ggctacgggc tagtttttac ctcttttaga acccactgta
541 aaagtccgtt gtaaagcccg ttctcactgt tggcgttttt ttttttttgg tttagtttct
601 tatttttcat ttttttcttt catgaccaaa aacaaacaaa tctcgcgatt tgtactgcgg
661 ccactggggc gtggccaaaa aaatgacaaa tttagaaacc ttagtttctg atttttcctg
721 ttatgaggag atatgataaa aaatattact gctttattgt ttttttttta tctactgaaa
781 tagagaaact tacccaagga ggaggcaaaa aaaagagtat atatacagca gctaccattc
841 agattttaat atattctttt ctcttcttct acactattat tataataatt ttactatatt
901 catttttagc ttaaaacctc atagaatatt attcttcagt cactcgctta aatacttatc
961 aaaaatgtct aagttggaaa aattcaccaa ctgttactct ttgtctaaga ctttgagatt
1021 caaggccatc ccagttggta agacccaaga aaacatcgac aacaagagac tattagttga
1081 agatgaaaag agagctgaag actacaaggg tgtcaagaaa ttgttggaca gatactactt
1141 gtcttttatc aacgacgttt tgcattccat caagctaaag aacttgaata actacatctc
1201 tttgttcaga aagaagacta gaactgaaaa ggaaaataag gaattggaaa acttggaaat
1261 caacttgaga aaggaaattg ctaaggcttt caagggtaat gaaggttaca agtctttatt
1321 caagaaagac atcattgaaa ccattttgcc agaatttttg gatgataagg atgaaattgc
1381 tttggttaac tctttcaacg gtttcaccac tgctttcact ggtttcttcg acaacagaga
1441 aaacatgttc tccgaggaag ctaaatccac ttctattgct ttcagatgta tcaacgaaaa
1501 cttgacccgt tacatctcta acatggacat ttttgaaaag gtcgacgcca tctttgacaa
1561 gcacgaagtc caagaaatca aggaaaagat cttaaactcc gactacgatg tcgaagattt
1621 cttcgaaggt gaattcttca actttgtttt aacccaagaa ggtatcgatg tctacaacgc
1681 cattatcggt ggttttgtca ctgaatctgg tgaaaagatc aagggtttga acgaatacat
1741 taacttgtac aaccaaaaga ccaaacaaaa attgccaaag ttcaagccat tgtacaagca
1801 agttttgtct gacagagaat ctttgtcttt ttacggtgaa gggtacacct ctgacgaaga
1861 agtcttggaa gtcttcagaa acactttgaa caagaactct gaaatcttct cctccatcaa
1921 gaagttagaa aagttgttca agaacttcga tgaatactct tctgctggta tcttcgttaa
1981 gaacggtcca gccatctcta ccatttctaa ggatatcttt ggtgaatgga acgtcattag
2041 agacaaatgg aacgctgaat acgatgacat ccatttgaag aaaaaggctg ttgtcaccga
2101 aaagtacgaa gacgacagaa gaaaatcctt caagaagatc ggttccttct ccttggaaca
2161 attacaagaa tacgccgatg ccgatttgtc cgttgtcgaa aaattgaagg aaattattat
2221 tcaaaaggtt gatgaaattt acaaagttta cggttcctct gaaaagttat tcgatgctga
2281 tttcgtcttg gaaaagtctt tgaagaagaa cgacgctgtt gtcgctatca tgaaggactt
2341 gttggactct gtcaaatctt tcgaaaacta tatcaaggcc ttcttcggtg aaggtaagga
2401 aactaacaga gatgaatcct tctacggtga ctttgtcttg gcttacgata ttttgttgaa
2461 ggttgaccac atctacgatg ccatcagaaa ctacgttact caaaagccat actctaagga
2521 caaattcaag ttgtacttcc aaaacccaca attcatgggt ggttgggata aggacaagga
2581 aactgactac agagctacca ttttgagata cggttccaag tactacttgg ccatcatgga
2641 caagaagtac gccaagtgtt tgcaaaagat tgacaaggac gatgtcaacg gtaactacga
2701 aaagattaac tacaagttgt tgccaggtcc aaacaagatg ttgccaaagg ttttcttctc
2761 caaaaagtgg atggcttact acaacccatc tgaagacatc caaaagatct acaagaacgg
2821 tactttcaaa aagggtgaca tgttcaactt aaacgactgt cacaagttga tcgacttctt
2881 caaggactcc atctctagat acccgaaatg gtccaacgct tacgatttca acttctctga
2941 aactgaaaaa tacaaggata ttgctggttt ctaccgtgaa gtcgaggaac aaggttataa
3001 ggtttctttc gaatccgctt ctaagaaaga agttgacaaa ttagtcgaag aaggtaagtt
3061 gtacatgttc caaatctaca acaaagattt ctccgacaag tctcacggta ctccaaactt
3121 gcacaccatg tacttcaagt tgctattcga tgaaaacaac cacggtcaaa tcagattgtc
3181 tggtggtgct gaattgttca tgagacgtgc ttctctaaag aaggaagaat tagtcgtcca
3241 cccagctaac tctccaattg ccaacaagaa cccagacaac cctaagaaga ccaccacttt
3301 gtcctacgac gtttacaagg acaagagatt ctccgaagac caatacgaat tgcacattcc
3361 aattgctatc aacaagtgtc caaagaacat cttcaagatc aacactgaag tcagagtttt
3421 gttaaagcac gatgacaacc cttacgttat tggtatcgac cgtggtgaaa gaaatttgtt
3481 gtacattgtt gttgttgacg gtaagggtaa catcgttgaa caatactcct tgaacgaaat
3541 catcaacaac ttcaacggta ttagaatcaa gactgattac cactctttgt tggataagaa
3601 ggaaaaggaa cgttttgaag ctcgtcaaaa ctggacctct attgaaaaca tcaaagaatt
3661 gaaggctggt tacatcagtc aagttgtcca caagatctgt gaattggtcg agaagtacga
3721 tgccgttatt gccttggaag atttgaactc tggttttaag aactctcgtg tcaaggttga
3781 aaagcaagtc taccaaaagt tcgaaaagat gttaatcgac aaattgaact acatggttga
3841 caagaaatcc aacccatgtg ctaccggtgg tgctttgaaa ggttaccaaa tcaccaacaa
3901 attcgaatct ttcaaatcta tgtccactca aaacgggttc atcttctaca ttccagcttg
3961 gttgacctcc aagatcgacc catctaccgg tttcgttaac ttgttgaaga ccaagtacac
4021 ttccattgct gattccaaga agttcatctc ttctttcgac agaatcatgt acgttccaga
4081 agaagacttg ttcgaattcg ccttggacta taagaacttc tccagaaccg atgctgacta
4141 cattaagaaa tggaaattgt actcctacgg taacagaatc agaattttca gaaacccaaa
4201 gaaaaacaac gttttcgatt gggaagaagt ttgtttgact tctgcctaca aggaattatt
4261 caacaaatac ggtatcaact accaacaagg tgatatcaga gctttgttgt gtgaacaatc
4321 tgacaaggct ttctactctt ccttcatggc tttgatgtcc ttgatgttgc aaatgagaaa
4381 ctccatcact ggtagaactg atgtcgactt cctcatttct ccagttaaga attctgacgg
4441 tattttctac gactctagaa attacgaagc tcaagaaaac gctattttgc caaagaacgc
4501 tgatgctaac ggtgcttaca atattgctag aaaggttttg tgggctatcg gtcaattcaa
4561 gaaggctgaa gacgaaaagc tagacaaggt caagattgct atttctaaca aggaatggtt
4621 ggaatacgct caaacctccg tcaagcactc cagagctgat ccaaagaaga agagaaaggt
4681 ataaaggagt taaaggcaaa gttttctttt ctagagccgt tcccacaaat aattatacgt
4741 atatgcttct tttcgtttac tatatatcta tatttacaag cctttattca ctgatgcaat
4801 ttgtttccaa atactttttt ggagatctca taactagata tcatgatggc gcaacttggc
4861 gctatcttaa ttactctggc tgccaggccc gtgtagaggg ccgcaagacc ttctgtacgc
4921 catatagtct ctaagaactt gaacaagttt ctagacctat tgccgccttt cggatcgcta
4981 ttgttgcggc cgccagctga agcttcgtac gctgcaggtc gacgaattct accgttcgta
5041 taatgtatgc tatacgaagt tatagatctg tttagcttgc ctcgtccccg ccgggtcacc
5101 cggccagcga catggaggcc cagaataccc tccttgacag tcttgacgtg cgcagctcag
5161 gggcatgatg tgactgtcgc ccgtacattt agcccataca tccccatgta taatcatttg
5221 catccataca ttttgatggc cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcaga
5281 cctgcgagca gggaaacgct cccctcacag acgcgttgaa ttgtccccac gccgcgcccc
5341 tgtagagaaa tataaaaggt taggatttgc cactgaggtt cttctttcat atacttcctt
5401 ttaaaatctt gctaggatac agttctcaca tcacatccga acataaacaa ccatgggtaa
5461 ggaaaagact cacgtttcga ggccgcgatt aaattccaac atggatgctg atttatatgg
5521 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gattgtatgg
5581 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
5641 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
5701 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggcaaaac
5761 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
5821 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
5881 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
5941 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataagct
6001 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
6061 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
6121 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
6181 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
6241 gatgctcgat gagtttttct aatcagtact gacaataaaa agattcttgt tttcaagaac
6301 ttgtcatttg tatagttttt ttatattgta gttgttctat tttaatcaaa tgttagcgtg
6361 atttatattt tttttcgcct cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa
6421 gtaatatcat gcgtcaatcg tatgtgaatg ctggtcgcta tactgctgtc gattcgatac
6481 taacgccgcc atccagtgtc gaaaacgagc tcataacttc gtataatgta tgctatacga
6541 acggtagaat tcgatatcag atccactagt ggcctatgcg gccgccaccg cggtggagct
6601 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
6661 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
6721 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
6781 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
6841 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
6901 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
6961 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
7021 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
7081 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
7141 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
7201 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
7261 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
7321 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
7381 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
7441 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
7501 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
7561 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
7621 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
7681 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
7741 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
7801 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
7861 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
7921 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
7981 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
8041 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
8101 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
8161 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
8221 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
8281 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
8341 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
8401 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
8461 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
8521 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
8581 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
8641 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
8701 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
8761 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgggtcc ttttcatcac
8821 gtgctataaa aataattata atttaaattt tttaatataa atatataaat taaaaataga
8881 aagtaaaaaa agaaattaaa gaaaaaatag tttttgtttt ccgaagatgt aaaagactct
8941 agggggatcg ccaacaaata ctacctttta tcttgctctt cctgctctca ggtattaatg
9001 ccgaattgtt tcatcttgtc tgtgtagaag accacacacg aaaatcctgt gattttacat
9061 tttacttatc gttaatcgaa tgtatatcta tttaatctgc ttttcttgtc taataaatat
9121 atatgtaaag tacgcttttt gttgaaattt tttaaacctt tgtttatttt tttttcttca
9181 ttccgtaact cttctacctt ctttatttac tttctaaaat ccaaatacaa aacataaaaa
9241 taaataaaca cagagtaaat tcccaaatta ttccatcatt aaaagatacg aggcgcgtgt
9301 aagttacagg caagcgatcc gtctaagaaa ccattattat catgacatta acctataaaa
9361 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
9421 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
9481 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
9541 catcagagca gattgtactg agagtgcacc atagatcacg acattactat atatataata
9601 taggaagcat ttaatagaac agcatcgtaa tatatgtgta ctttgcagtt atgacgccag
9661 atggcagtag tggaagatat tctttattga aaaatagctt gtcaccttac gtacaatctt
9721 gatccggagc ttttcttttt ttgccgatta agaattaatt cggtcgaaaa aagaaaagga
9781 gagggccaag agggagggca ttggtgacta ttgagcacgt gagtatacgt gattaagcac
9841 acaaaggcag cttggagtat gtctgttatt aatttcacag gtagttctgg tccattggtg
9901 aaagtttgcg gcttgcagag cacagaggcc gcagaatgtg ctctagattc cgatgctgac
9961 ttgctgggta ttatatgtgt gcccaataga aagagaacaa ttgacccggt tattgcaagg
10021 aaaatttcaa gtcttgtaaa agcatataaa aatagttcag gcactccgaa atacttggtt
10081 ggcgtgtttc gtaatcaacc taaggaggat gttttggctc tggtcaatga ttacggcatt
10141 gatatcgtcc aactgcatgg agatgagtcg tggcaagaat accaagagtt cctcggtttg
10201 ccagttatta aaagactcgt atttccaaaa gactgcaaca tactactcag tgcagcttca
10261 cagaaacctc attcgtttat tcccttgttt gattcagaag caggtgggac aggtgaactt
10321 ttggattgga actcgatttc tgactgggtt ggaaggcaag agagccccga aagcttacat
10381 tttatgttag ctggtggact gacgccagaa aatgttggtg atgcgcttag attaaatggc
10441 gttattggtg ttgatgtaag cggaggtgtg gagacaaatg gtgtaaaaga ctctaacaaa
10501 atagcaaatt tcgtcaaaaa tgctaagaaa taggttatta ctgagtagta tttatttaag
10561 tattgtttgt gcacttgccg atctatgcgg tgtgaaatac cgcacagatg cgtaaggaga
10621 aaataccgca tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
10681 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
10741 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
10801 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
10861 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
10921 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
10981 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
11041 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgcc
11101 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
11161 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
11221 gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga
11281 attgggtacc ttttcttttt ttgcggtcac ccccatgtgg cgggga
//
