| 质粒名称: | pLX_311-Cas9 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 96924 |
| 存储实验室: | John Doench, William Hahn, David Root |
|
载体骨架基本信息 |
|
| 载体名称: | pXPR |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，慢病毒 |
| 筛选标记: | 杀稻瘟菌素 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称: | Cas9 |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | EF1a |
| 标签/融合蛋白: |
3xFlag
V5
|
| 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: | pLX_311-Cas9 was a gift from John Doench & William Hahn & David Root (Addgene plasmid # 96924 ; http://n2t.net/addgene:96924 ; RRID:Addgene_96924) |
| 参考文献部分: | Rational design of highly active sgRNAs for CRISPR-Cas9-mediated gene inactivation. Doench JG, Hartenian E, Graham DB, Tothova Z, Hegde M, Smith I, Sullender M, Ebert BL, Xavier RJ, Root DE. Nat Biotechnol. 2014 Sep 3. doi: 10.1038/nbt.3026. 10.1038/nbt.3026 PubMed 25184501 |
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 pLX_311-Cas9 12494 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION for Cas9 knockout, lentiviral expression of Cas9. Alternate plasmid
name: pXR111.
ACCESSION .
VERSION .
KEYWORDS pLX_311-Cas9
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12494)
AUTHORS Doench JG, Hartenian E, Graham DB, Tothova Z, Hegde M, Smith I,
Sullender M, Ebert BL, Xavier RJ, Root DE
TITLE Rational design of highly active sgRNAs for CRISPR-Cas9-mediated
gene inactivation.
JOURNAL Nat Biotechnol. 2014 Sep 3. doi: 10.1038/nbt.3026.
PUBMED 25184501
REFERENCE 2 (bases 1 to 12494)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..12494
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(30..47)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(201..789)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(281..300)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(960..1820)
/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 1583..1602
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1821..1925)
/gene="bla"
/label=AmpR promoter
rep_origin complement(1951..2406)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2088..2109)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2300..2319
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2533..2555
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 2547..2564
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 2548..2564
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2574..2593
/label=T7
/note="T7 promoter, forward primer"
promoter 2574..2592
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
rep_origin 2613..2748
/label=SV40 ori
/note="SV40 origin of replication"
polyA_signal 2788..2909
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2799..2818)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2853..2872
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
LTR 2987..3220
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
misc_feature 3292..3880
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS 3398..3409
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind 3807..3827
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(3931..3972)
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
protein_bind 3980..4004
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS complement(4007..4054)
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS complement(4055..8155)
/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 complement(8180..8200)
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS complement(8207..8272)
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
protein_bind complement(8275..8299)
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
promoter complement(8319..9497)
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 8328..9266
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind complement(8351..8371)
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
misc_feature 9635..9752
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
CDS complement(9826..10221)
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/label=BSD
/note="confers resistance to blasticidin"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
promoter complement(10243..10439)
/label=SV40 promoter
/note="SV40 early promoter"
promoter complement(10251..10461)
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 10257..10392
/label=SV40 ori
/note="SV40 origin of replication"
CDS complement(10692..10736)
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 10921..11154
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 11647..11772
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
LTR 11819..11999
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
promoter complement(12000..12226)
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
primer_bind complement(12254..12274)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(12254..12272)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(12293..12309)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(12293..12309)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(12306..12328)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 12317..12333
/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(12341..12371)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 12386..12407
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
ORIGIN
1 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
61 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
121 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
181 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
241 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
301 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
361 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
421 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
481 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
541 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
601 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
661 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
721 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
781 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
841 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
901 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
961 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
1021 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
1081 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
1141 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
1201 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
1261 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
1321 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
1381 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
1441 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
1501 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
1561 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
1621 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
1681 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
1741 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
1801 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
1861 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
1921 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
1981 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
2041 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
2101 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
2161 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
2221 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
2281 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
2341 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
2401 gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
2461 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
2521 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg
2581 actcactata gggcgaattg ggtacgtccc taggcctcca aaaaagcctc ctcactactt
2641 ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa aaattagtca
2701 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag
2761 gggcgggata gctagagcca gacatgataa gatacattga tgagtttgga caaaccacaa
2821 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
2881 taaccattat aagctgcaat aaacaagttc ctctcactct ctgatattca tttctttgca
2941 agttataaat actgaataat aagatgacat gaactactat acgtactgct agagattttc
3001 cacactgact aaaagggtct gagggatctc tagttaccag agtcacacaa cagacgggca
3061 cacactactt gaagcactca aggcaagctt tattgaggct taagcagtgg gttccctagt
3121 tagccagaga gctcccaggc tcagatctgg tctaaccaga gagacccagt acaagcaaaa
3181 agcagatctt gtcttcgttg ggagtgaatt agcccttcca gtcccccctt ttcttttaaa
3241 aagtggctaa gatctacagc tgccttgtaa gtcattggtc ttaaagtcga cgcggggagg
3301 cggcccaaag ggagatccga ctcgtctgag ggcgaaggcg aagacgcgga agaggccgca
3361 gagccggcag caggccgcgg gaaggaaggt ccgctggatt gagggccgaa gggacgtagc
3421 agaaggacgt cccgcgcaga atccaggtgg caacacaggc gagcagccaa ggaaaggacg
3481 atgatttccc cgacaacacc acggaattgt cagtgcccaa cagccgagcc cctgtccagc
3541 agcgggcaag gcaggcggcg atgagttccg ccgtggcaat agggaggggg aaagcgaaag
3601 tcccggaaag gagctgacag gtggtggcaa tgccccaacc agtgggggtt gcgtcagcaa
3661 acacagtgca caccacgcca cgttgcctga caacgggcca caactcctca taaagagaca
3721 gcaaccagga tttatacaag gaggagaaaa tgaaagccat acgggaagca atagcatgat
3781 acaaaggcat taaagcagcg tatccacata gcgtaaaagg agcaacatag ttaagaatac
3841 cagtcaatct ttcacaaatt ttgtaatcca gaggttgatt gtcgacttaa cgcgttgatt
3901 gtcgacttaa ccggtactag ttcattacta cgtagaatcg agaccgagga gagggttagg
3961 gataggctta ccgatatcaa ccactttgta caagaaagtt gggggtcttt ttcttttttg
4021 cctggccggc ctttttcgtg gccgccggcc ttttgtcgcc tcccagctga gacaggtcga
4081 tccgtgtctc gtacaggccg gtgatgctct ggtggatcag ggtggcgtcc agcacctctt
4141 tggtgctggt gtacctcttc cggtcgatgg tggtgtcaaa gtacttgaag gcggcagggg
4201 ctcccagatt ggtcagggta aacaggtgga tgatattctc ggcctgctct ctgatgggct
4261 tatcccggtg cttgttgtag gcggacagca ctttgtccag attagcgtcg gccaggatca
4321 ctctcttgga gaactcgctg atctgctcga tgatctcgtc caggtagtgc ttgtgctgtt
4381 ccacaaacag ctgtttctgc tcattatcct cgggggagcc cttcagcttc tcatagtggc
4441 tggccaggta caggaagttc acatatttgg agggcagggc cagttcgttt cccttctgca
4501 gttcgccggc agaggccagc attctcttcc ggccgttttc cagctcgaac agggagtact
4561 taggcagctt gatgatcagg tcctttttca cttctttgta gcccttggct tccagaaagt
4621 cgatgggatt cttctcgaag ctgcttcttt ccatgatggt gatccccagc agctctttca
4681 cactcttcag tttcttggac ttgccctttt ccactttggc caccaccagc acagaatagg
4741 ccacggtggg gctgtcgaag ccgccgtact tcttagggtc ccagtccttc tttctggcga
4801 tcagcttatc gctgttcctc ttgggcagga tagactcttt gctgaagccg cctgtctgca
4861 cctcggtctt tttcacgata ttcacttggg gcatgctcag cactttccgc acggtggcaa
4921 aatcccggcc cttatcccac acgatctccc cggtttcgcc gtttgtctcg atcagaggcc
4981 gcttccggat ctcgccgttg gccagggtaa tctcggtctt gaaaaagttc atgatgttgc
5041 tgtagaagaa gtacttggcg gtagccttgc cgatttcctg ctcgctcttg gcgatcatct
5101 tccgcacgtc gtacaccttg tagtcgccgt acacgaactc gctttccagc ttagggtact
5161 ttttgatcag ggcggttccc acgacggcgt tcaggtaggc gtcgtgggcg tggtggtagt
5221 tgttgatctc gcgcactttg taaaactgga aatccttccg gaaatcggac accagcttgg
5281 acttcagggt gatcactttc acttcccgga tcagcttgtc attctcgtcg tacttagtgt
5341 tcatccggga gtccaggatc tgtgccacgt gctttgtgat ctgccgggtt tccaccagct
5401 gtctcttgat gaagccggcc ttatccagtt cgctcaggcc gcctctctcg gccttggtca
5461 gattgtcgaa ctttctctgg gtaatcagct tggcgttcag cagctgccgc cagtagttct
5521 tcatcttctt cacgacctct tcggagggca cgttgtcgct cttgccccgg ttcttgtcgc
5581 ttctggtcag caccttgttg tcgatggagt cgtccttcag aaagctctga ggcacgatat
5641 ggtccacatc gtagtcggac agccggttga tgtccagttc ctggtccacg tacatatccc
5701 gcccattctg caggtagtac aggtacagct tctcgttctg cagctgggtg ttttccacgg
5761 ggtgttcttt caggatctgg ctgcccagct ctttgatgcc ctcttcgatc cgcttcattc
5821 tctcgcggct gttcttctgt cccttctggg tggtctggtt ctctctggcc atttcgatca
5881 cgatgttctc gggcttgtgc cggcccatca ctttcacgag ctcgtccacc accttcactg
5941 tctgcaggat gcccttctta atggcggggc tgccggccag attggcaatg tgctcgtgca
6001 ggctatcgcc ctggccggac acctgggctt tctggatgtc ctctttaaag gtcaggctgt
6061 cgtcgtggat cagctgcatg aagtttctgt tggcgaagcc gtcggacttc aggaaatcca
6121 ggattgtctt gccggactgc ttgtcccgga tgccgttgat cagcttccgg ctcagcctgc
6181 cccagccggt gtatctccgc cgcttcagct gcttcatcac tttgtcgtcg aacaggtggg
6241 cataggtttt cagccgttcc tcgatcatct ctctgtcctc aaacagtgtc agggtcagca
6301 cgatatcttc cagaatgtcc tcgttttcct cattgtccag gaagtccttg tccttgataa
6361 ttttcagcag atcgtggtat gtgcccaggg aggcgttgaa ccgatcttcc acgccggaga
6421 tttccacgga gtcgaagcac tcgattttct tgaagtagtc ctctttcagc tgcttcacgg
6481 tcactttccg gttggtcttg aacagcaggt ccacgatggc ctttttctgc tcgccgctca
6541 ggaaggcggg ctttctcatt ccctcggtca cgtatttcac tttggtcagc tcgttataca
6601 cggtgaagta ctcgtacagc aggctgtgct tgggcagcac cttctcgttg ggcaggttct
6661 tatcgaagtt ggtcatccgc tcgatgaagc tctgggcgga agcgcccttg tccaccactt
6721 cctcgaagtt ccagggggtg atggtttcct cgctctttct ggtcatccag gcgaatctgc
6781 tgtttcccct ggccagaggg cccacgtagt aggggatgcg gaaggtcagg atcttctcga
6841 tcttttcccg gttgtccttc aggaatgggt aaaaatcttc ctgccgccgc agaatggcgt
6901 gcagctctcc caggtggatc tggtggggga tgctgccgtt gtcgaaggtc cgctgcttcc
6961 gcagcaggtc ctctctgttc agcttcacga gcagttcctc ggtgccgtcc atcttttcca
7021 ggatgggctt gatgaacttg tagaactctt cctggctggc tccgccgtca atgtagccgg
7081 cgtagccgtt cttgctctgg tcgaagaaaa tctctttgta cttctcaggc agctgctgcc
7141 gcacgagagc tttcagcagg gtcaggtcct ggtggtgctc gtcgtatctc ttgatcatag
7201 aggcgctcag gggggccttg gtgatctcgg tgttcactct caggatgtcg ctcagcagga
7261 tggcgtcgga caggttcttg gcggccagaa acaggtcggc gtactggtcg ccgatctggg
7321 ccagcaggtt gtccaggtcg tcgtcgtagg tgtccttgct cagctgcagt ttggcatcct
7381 cggccaggtc gaagttgctc ttgaagttgg gggtcaggcc caggctcagg gcaatcaggt
7441 ttccgaacag gccattcttc ttctcgccgg gcagctgggc gatcagattt tccagccgtc
7501 tgctcttgct cagtctggca gacaggatgg ccttggcgtc cacgccgctg gcgttgatgg
7561 ggttttcctc gaacagctgg ttgtaggtct gcaccagctg gatgaacagc ttgtccacgt
7621 cgctgttgtc ggggttcagg tcgccctcga tcaggaagtg gccccggaac ttgatcatgt
7681 gggccagggc cagatagatc agccgcaggt cggccttgtc ggtgctgtcc accagtttct
7741 ttctcaggtg gtagatggtg gggtacttct cgtggtaggc cacctcgtcc acgatgttgc
7801 cgaagatggg gtgccgctcg tgcttcttat cctcttccac caggaaggac tcttccagtc
7861 tgtggaagaa gctgtcgtcc accttggcca tctcgttgct gaagatctct tgcagatagc
7921 agatccggtt cttccgtctg gtgtatcttc ttctggcggt tctcttcagc cgggtggcct
7981 cggctgtttc gccgctgtcg aacagcaggg ctccgatcag gttcttcttg atgctgtgcc
8041 ggtcggtgtt gcccagcacc ttgaatttct tgctgggcac cttgtactcg tcggtgatca
8101 cggcccagcc cacagagttg gtgccgatgt ccaggccgat gctgtacttc ttgtcggctg
8161 ctgggactcc gtggataccg accttccgct tcttctttgg ggccatctta tcgtcatcgt
8221 ctttgtaatc aatatcatga tccttgtagt ctccgtcgtg gtccttatag tccatgccaa
8281 cttttttgta caaacttgtt gatcaatcga tgctagcctc acgacacctg aaatggaaga
8341 aaaaaacttt gaaccactgt ctgaggcttg agaatgaacc aagatccaaa ctcaaaaagg
8401 gcaaattcca aggagaatta catcaagtgc caagctggcc taacttcagt ctccacccac
8461 tcagtgtggg gaaactccat cgcataaaac ccctcccccc aacctaaaga cgacgtactc
8521 caaaagctcg agaactaatc gaggtgcctg gacggcgccc ggtactccgt ggagtcacat
8581 gaagcgacgg ctgaggacgg aaaggccctt ttcctttgtg tgggtgactc acccgcccgc
8641 tctcccgagc gccgcgtcct ccattttgag ctccctgcag cagggccggg aagcggccat
8701 ctttccgctc acgcaactgg tgccgaccgg gccagccttg ccgcccaggg cggggcgata
8761 cacggcggcg cgaggccagg caccagagca ggccggccag cttgagacta cccccgtccg
8821 attctcggtg gccgcgctcg caggccccgc ctcgccgaac atgtgcgctg ggacgcacgg
8881 gccccgtcgc cgcccgcggc cccaaaaacc gaaataccag tgtgcagatc ttggcccgca
8941 tttacaagac tatcttgcca gaaaaaaagc gtcgcagcag gtcatcaaaa attttaaatg
9001 gctagagact tatcgaaagc agcgagacag gcgcgaaggt gccaccagat tcgcacgcgg
9061 cggccccagc gcccaggcca ggcctcaact caagcacgag gcgaaggggc tccttaagcg
9121 caaggcctcg aactctccca cccacttcca acccgaagct cgggatcaag aatcacgtac
9181 tgcagccagg tggaagtaat tcaaggcacg caagggccat aacccgtaaa gaggccaggc
9241 ccgcgggaac cacacacggc acttacctgt gttctggcgg caaacccgtt gcgaaaaaga
9301 acgttcacgg cgactactgc acttatatac ggttctcccc caccctcggg aaaaaggcgg
9361 agccagtaca cgacatcact ttcccagttt accccgcgcc accttctcta ggcaccggtt
9421 caattgccga cccctccccc caacttctcg gggactgtgg gcgatgtgcg ctctgcccac
9481 tgacgggcac cggagccaat tcccactcct ttcaagacct agaaggtcca ttagctgcaa
9541 agattcctct ctgtttaaaa ctttatccat ctttgcacct gcaggcctcg agccggcgcc
9601 aaagtggatc tctgctgtcc ctgtaataaa cccgaaaatt ttgaattttt gtaatttgtt
9661 tttgtaattc tttagtttgt atgtctgttg ctattatgtc tactattctt tcccctgcac
9721 tgtacccccc aatcccccct tttcttttaa aattgtggat gaatactgcc atttgtctca
9781 agatctagaa ttcgttaacg gcgcgcctta ctactgcagc tgcaggccct cccacacata
9841 accagagggc agcaattcac gaatcccaac tgccgtcggc tgtccatcac tgtccttcac
9901 tatggctttg atcccaggat gcagatcgag aagcacctgt cggcaccgtc cgcaggggct
9961 caagatgccc ctgttctcat ttccgatcgc gacgatacaa gtcaggttgc cagctgccgc
10021 agcagcagca gtgcccagca ccacgagttc tgcacaaggt cccccagtaa aatgatatac
10081 attgacacca gtgaagatgc ggccgtcgct agagagagct gcgctggcga cgctgtagtc
10141 ttcagagatg gggatgctgt tgattgtagc cgttgctctt tcaatgaggg tggattcttc
10201 ttgagacaaa ggcttggcca tggtggtacc tctagaaagc tttttgcaaa agcctaggcc
10261 tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc
10321 tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt
10381 taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat tgagacgcat
10441 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta
10501 attgagatgc aatgcatggg tcccctcggg gttgggaggt gggtctgaaa cgataatggt
10561 gaatatccct gcctaactct attcactata gaaagtacag caaaaactat tcttaaacct
10621 accaagcctc ctactatcat tatgaataat tttatatacc acagccaatt tgttatgtta
10681 aaccaattcc acaaacttgc ccatttatct aattccaata attcttgttc attcttttct
10741 tgctggtttt gcgattcttc aattaaggag tgtattaagc ttgtgtaatt gttaatttct
10801 ctgtcccact ccatccaggt cgtgtgattc caaatctgtt ccagagattt attactccaa
10861 ctagcattcc aaggcacagc agtggtgcaa atgagttttc cagagcaacc ccaaatcccc
10921 aggagctgtt gatcctttag gtatctttcc acagccagga ttcttgcctg gagctgcttg
10981 atgccccaga ctgtgagttg caacagatgc tgttgcgcct caatagccct cagcaaattg
11041 ttctgctgct gcactatacc agacaataat tgtctggcct gtaccgtcag cgtcattgac
11101 gctgcgccca tagtgcttcc tgctgctccc aagaacccaa ggaacaaagc tcctattccc
11161 actgctcttt tttctctctg caccactctt ctctttgcct tggtgggtgc tactcctaat
11221 ggttcaattt ttactacttt atatttatat aattcacttc tccaattgtc cctcatatct
11281 cctcctccag gtctgaagat cagcggccgc ttgctgtgcg gtggtcttac ttttgttttg
11341 ctcttcctct atcttgtcta aagcttcctt ggtgtctttt atctctatcc tttgatgcac
11401 acaatagagg gttgctactg tattatataa tgatctaagt tcttctgatc ctgtctgaag
11461 ggatggttgt agctgtccca gtatttgtct acagccttct gatgtttcta acaggccagg
11521 attaactgcg aatcgttcta gctccctgct tgcccatact atatgtttta atttatattt
11581 tttctttccc cctggcctta accgaatttt ttcccatcgc gatctaattc tcccccgctt
11641 aatactgacg ctctcgcacc catctctctc cttctagcct ccgctagtca aaatttttgg
11701 cgtactcacc agtcgccgcc cctcgcctct tgccgtgcgc gcttcagcaa gccgagtcct
11761 gcgtcgagag agctcctctg gtttcccttt cgctttcaag tccctgttcg ggcgccactg
11821 ctagagattt tccacactga ctaaaagggt ctgagggatc tctagttacc agagtcacac
11881 aacagacggg cacacactac ttgaagcact caaggcaagc tttattgagg cttaagcagt
11941 gggttcccta gttagccaga gagctcccag gctcagatct ggtctaacca gagagacccg
12001 tttatgtatc gagctaggca cttaaataca atatctctgc aatgcggcaa ttcagtggtt
12061 cgtccaatcc atgtcagacc cgtctgttgc cttcctaata aggcacgatc gtaccacctt
12121 acttccacca atcggcatgc acggtgcttt ttctctcctt gtaaggcatg ttgctaactc
12181 atcgttacca tgttgcaaga ctacaagagt attgcataag actacattaa gcttgcagct
12241 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
12301 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
12361 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
12421 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
12481 gcgcggggag aggc
//
