| 质粒名称: | pCas9-CAT |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 80323 |
| 存储实验室: | Sebastian Lourido |
|
载体骨架基本信息 |
|
| 载体名称: | pU6-Universal |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 8360 bp |
| 完整质粒大小: | 9301 bp |
| 载体修饰: | sgRNA区域已被氯霉素乙酰转移酶替换为TUB1 5’UTR和SAG1 3’UTR |
| 载体类型: | CRISPR |
| 筛选标记: | CAT |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Cas9 |
| 物种来源: | Streptococcus pyogenes |
| 插入大小: | 4101 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | TgTUB1 |
| 标签/融合蛋白: |
3X标志（插入件上的N端子）
NLS（插入件上的N端子）
NLS（插接件上的C端子）
|
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | BGH-rev |
|
文献引用方法 |
|
| 材料和方法部分: | pCas9-CAT was a gift from Sebastian Lourido (Addgene plasmid # 80323 ; http://n2t.net/addgene:80323 ; RRID:Addgene_80323) |
| 参考文献部分: | A Genome-wide CRISPR Screen in Toxoplasma Identifies Essential Apicomplexan Genes. Sidik SM, Huet D, Ganesan SM, Huynh MH, Wang T, Nasamu AS, Thiru P, Saeij JP, Carruthers VB, Niles JC, Lourido S. Cell. 2016 Sep 8;166(6):1423-1435.e12. doi: 10.1016/j.cell.2016.08.019. Epub 2016 Sep 2. 10.1016/j.cell.2016.08.019 PubMed 27594426 |
买家导航
pCas9-CAT载体 pCas9CAT质粒基本信息
pCas9-CAT质粒图谱 pCas9CAT载体图谱和pCas9-CAT载体序列 pCas9CAT质粒序列多克隆位点信息
pCas9-CAT质粒 pCas9CAT载体简介
pCas9-CAT质粒序列 pCas9CAT载体序列
LOCUS pCas9-CAT 9301 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Encodes Cas9 and chloramphenicol acetyltransferase (CAT).
ACCESSION .
VERSION .
KEYWORDS pCas9-CAT
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9301)
AUTHORS Sidik SM, Huet D, Ganesan SM, Huynh MH, Wang T, Nasamu AS, Thiru P,
Saeij JP, Carruthers VB, Niles JC, Lourido S
TITLE A Genome-wide CRISPR Screen in Toxoplasma Identifies Essential
Apicomplexan Genes.
JOURNAL Cell. 2016 Sep 8;166(6):1423-1435.e12. doi:
10.1016/j.cell.2016.08.019. Epub 2016 Sep 2.
PUBMED 27594426
REFERENCE 2 (bases 1 to 9301)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9301
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 509..574
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 581..601
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 626..4726
/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 4727..4774
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
primer_bind complement(4802..4819)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 4808..5015
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 5024..5164
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 5024..5153
/label=AAV2 ITR
rep_origin 5239..5694
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5326..5345)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5536..5557
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(5711..5730)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5830..5852
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(5890..5908)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5976..6080
/gene="bla"
/label=AmpR promoter
CDS 6081..6941
/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 complement(6299..6318)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7112..7700
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7601..7620
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7773..7789
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind 7774..7790
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
CDS 8305..8967
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MHEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAF
LKTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFS
SLWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVAN
MDNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(8374..8393)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
primer_bind complement(9285..9301)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(9285..9301)
/label=pBluescriptSK
/note="For pBluescript vector"
ORIGIN
1 ggtaccatgc atgtcccgcg ttcgtgaaat tctctgcatc agcggagtga tcaggaatca
61 tcgtctcagc gggatgacgt tgcggagcag gccggctcgc ggtgggcagt cagatgccga
121 aggcgtaact caggacggct tgcgctcatc gcagaacagg ggtggtgcct gcattgggtg
181 cggttggtga tcctggttgg accggtggag atgcgcgcgc acgaagggga tgtgtcagaa
241 acattttgtt tgttctctgt gaacttttag atgtgttaaa ggcggcgaat attancagag
301 agtcctcctt gttggattct ctcttgaatt tcgccctttc cttctctttg cgagtctcgt
361 agagaacaag cactcgttcg ccgtccctga cgacgcaacc cgcgcagaag acatccacca
421 aacggtgtta cacaatcacc ttgtgtgaag ttcttgcgga aaactactcg ttggcatttt
481 ttcttgaatt ccctttttcg acaccatgga ctataaggac cacgacggag actacaagga
541 tcatgatatt gattacaaag acgatgacga taagatggcc ccaaagaaga agcggaaggt
601 cggtatccac ggagtcccag cagccgacaa gaagtacagc atcggcctgg acatcggcac
661 caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa
721 ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag ccctgctgtt
781 cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac
841 cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt
901 ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca
961 cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc
1021 caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct
1081 gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga
1141 cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa
1201 ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc
1261 tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc ccggcgagaa
1321 gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc ccaacttcaa
1381 gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga
1441 cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc
1501 caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac
1561 caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac
1621 cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga
1681 ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg aagagttcta
1741 caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct
1801 gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca
1861 gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct
1921 gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg
1981 ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat
2041 caccccctgg aacttcgagg aagtggtgga caagggcgct tccgcccaga gcttcatcga
2101 gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct
2161 gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg
2221 aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt
2281 caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga
2341 gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac
2401 ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga
2461 ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga
2521 ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg
2581 gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa
2641 gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt
2701 catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt
2761 gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat
2821 taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg
2881 gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg
2941 acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag
3001 ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct
3061 gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct
3121 gtccgactac gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg actccatcga
3181 caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg tgccctccga
3241 agaggtcgtg aagaagatga agaactactg gcggcagctg ctgaacgcca agctgattac
3301 ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa
3361 ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca
3421 gatcctggac tcccggatga acactaagta cgacgagaat gacaagctga tccgggaagt
3481 gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta
3541 caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt
3601 gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggcgacta
3661 caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg gcaaggctac
3721 cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc
3781 caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt
3841 gtgggataag ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa
3901 tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa
3961 gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg
4021 cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa
4081 gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag
4141 cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga
4201 cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat
4261 gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt
4321 gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga
4381 gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat
4441 cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc
4501 ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt
4561 taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg
4621 gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac
4681 cggcctgtac gagacacgga tcgacctgtc tcagctggga ggcgacaaaa ggccggcggc
4741 cacgaaaaag gccggccagg caaaaaagaa aaagtaagaa ttcctagagc tcgctgatca
4801 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
4861 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
4921 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
4981 gaggattggg aagagaatag caggcatgct ggggagcggc cgcaggaacc cctagtgatg
5041 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
5101 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
5161 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
5221 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
5281 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
5341 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
5401 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
5461 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
5521 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
5581 gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
5641 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
5701 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
5761 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
5821 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
5881 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
5941 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
6001 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
6061 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
6121 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
6181 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
6241 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
6301 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
6361 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
6421 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
6481 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
6541 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
6601 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
6661 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
6721 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
6781 gagccggtga gcgtggaagc cgcggtatca ttgcagcact ggggccagat ggtaagccct
6841 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
6901 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
6961 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
7021 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
7081 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
7141 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
7201 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
7261 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
7321 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
7381 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
7441 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
7501 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
7561 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
7621 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
7681 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
7741 gctggccttt tgctcacatg tcgggccccc cctcgaggtc gacggtatcg ataagcttga
7801 tggcgatgca tgtcccgcgt tcgtgaaatt ctctgcatca gcggagtgat caggaatcat
7861 cgtctcagcg ggatgacgtt gcggagcagg ccggctcgcg gtgggcagtc agatgccgaa
7921 ggcgtaactc aggacggctt gcgctcatcg cagaacaggg gtggtgcctg cattgggtgc
7981 ggttggtgat cctggttgga ccggtggaga tgcgcgcgca cgaaggggat gtgtcagaaa
8041 cattttgttt gttctctgtg aacttttaga tgtgttaaag gcggcgaata ttagcagaga
8101 gtcctccttg ttggattctc tcttgaattt cgccctttcc ttctctttgc gagtctcgta
8161 gagaacaagc actcgttcgc cgtccctgac gacgcaaccc gcgcagaaga catccaccaa
8221 acggtgttac acaatcacct tgtgtgaagt tcttgcggaa aactactcgt tggcattttt
8281 tcttgaattc cctttttcga caaaatgcat gagaaaaaaa tcactggata taccaccgtt
8341 gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt
8401 acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat
8461 aagcacaagt tttatccggc atttattcac attcttgccc gcctgatgaa tgctcatccg
8521 gaattccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt
8581 tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac
8641 gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg
8701 gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg
8761 agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc
8821 accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt
8881 catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac
8941 tgcgatgagt ggcagggcgg ggcttaatta atcaccgttg tgctcacttc tcaaatcgac
9001 aaaggaaaca cacttcgtgc agcatgtgcc ccattataaa gaaactgagt tgttccgttg
9061 tggcttgcag gtgtcacatc cacaaaaacc ggccgactct aaataggagt gtttcgcagc
9121 aagcagcgaa agtttatgac tgggtccgaa tctctgaacg gatgtgtggc ggacctggct
9181 gatgttgatc gccgtcgaca cacgcgccac atgggtcaat acacaagaca gctatcagtt
9241 gttttagtcg aaccggttaa cacaattctt gcccccccga gggggatcca ctagttctag
9301 a
//
