| 质粒名称: | Fuw-dCas9-Tet1CD |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 84475 |
| 存储实验室: | Rudolf Jaenisch |
|
载体骨架基本信息 |
|
| 载体名称: | FUW |
| 空载体大小: | 9213 bp |
| 完整质粒大小: | 15573 bp |
| 载体类型: | 哺乳细胞表达，慢病毒 |
| 筛选标记: | Zeo筛选标记，位于LTR区外面，不会被包入病毒。 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | dCas9-Tet1CD |
| 物种来源: | H. sapiens (human), Synthetic |
| 插入大小: | 6351 bp |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | AscI (未破坏) |
| 3’克隆位点: | EcoRI (未破坏) |
| 5’测序引物 | aactatgcgctcggg |
| 3’测序引物 | taaagcagcgtatcc |
|
文献引用方法 |
|
| 材料和方法部分: |
Fuw-dCas9-Tet1CD was a gift from Rudolf Jaenisch (Addgene plasmid # 84475 ; http://n2t.net/addgene:84475 ; RRID:Addgene_84475)
For viral preps, please replace (Addgene plasmid # 84475) in the above sentence with: (Addgene viral prep # 84475-LV) or (Addgene viral prep # 84475-LVC) |
| 参考文献部分: |
Editing DNA Methylation in the Mammalian Genome. Liu XS, Wu H, Ji X, Stelzer Y, Wu X, Czauderna S, Shu J, Dadon D, Young RA, Jaenisch R. Cell. 2016 Sep 22;167(1):233-247.e17. doi: 10.1016/j.cell.2016.08.056. 10.1016/j.cell.2016.08.056 PubMed 27662091 |
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 Fuw-dCas9-Tet1CD 15501 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Lentiviral construct to express dCas9 fused with the catalytic
domain of Tet1.
ACCESSION .
VERSION .
KEYWORDS Fuw-dCas9-Tet1CD
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 15501)
AUTHORS Liu XS, Wu H, Ji X, Stelzer Y, Wu X, Czauderna S, Shu J, Dadon D,
Young RA, Jaenisch R
TITLE Editing DNA Methylation in the Mammalian Genome.
JOURNAL Cell. 2016 Sep 22;167(1):233-247.e17. doi:
10.1016/j.cell.2016.08.056.
PUBMED 27662091
REFERENCE 2 (bases 1 to 15501)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..15501
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 12..33
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(150..167)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(321..909)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(401..420)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1080..1940)
/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 1703..1722
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1941..2045)
/gene="bla"
/label=AmpR promoter
primer_bind complement(2120..2139)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 2311..2690
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2692..2890
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2845..2865
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 2908..3088
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 3135..3260
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 3753..3986
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 4171..4215
/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 4513..4630
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 4695..5906
/label=UbC promoter
/note="human ubiquitin C promoter"
primer_bind 5743..5762
/label=hUBCpro-F
/note="Human Ubiquitin C (UbC) promoter, forward primer"
CDS 5963..10066
/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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
CDS 10082..10102
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 10106..10126
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 10130..10150
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
misc_feature 12338..12926
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(12391..12411)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(12809..12820)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(12929..12945)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(12930..12946)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 13451..13631
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
primer_bind complement(13657..13674)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 13663..13887
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 13933..14361
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(14020..14039)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 14230..14251
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(14370..14390)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 14414..14624
/label=SV40 promoter
/note="SV40 early promoter"
promoter 14436..14632
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 14483..14618
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 14545..14564
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 14680..14727
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 14746..15120
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=BleoR
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
polyA_signal 15250..15371
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(15287..15306)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 15341..15360
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(15420..15436)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(15420..15436)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(15433..15455)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 15444..15460
/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(15468..15498)
/label=lac promoter
/note="promoter for the E. coli lac operon"
ORIGIN
1 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
61 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
121 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
181 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
241 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
301 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
361 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
421 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
481 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
541 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
601 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
661 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
721 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
781 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
841 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
901 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
961 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
1021 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
1081 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
1141 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
1201 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
1261 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
1321 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
1381 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
1441 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
1501 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
1561 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
1621 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
1681 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
1741 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
1801 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
1861 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
1921 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
1981 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
2041 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
2101 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
2161 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
2221 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
2281 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt
2341 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
2401 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
2461 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
2521 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
2581 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
2641 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
2701 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
2761 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
2821 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
2881 atataagcag cgcgttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg
2941 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
3001 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
3061 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
3121 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
3181 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
3241 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
3301 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
3361 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
3421 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
3481 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
3541 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
3601 ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat
3661 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga
3721 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga
3781 gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta
3841 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat
3901 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa
3961 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc
4021 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat
4081 cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc
4141 ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat
4201 aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta
4261 ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata
4321 gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg
4381 aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga cagagacaga
4441 tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac aaatggcagt
4501 attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat
4561 agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat
4621 tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttaa ttaacccgtg
4681 tcggctccag atctggcctc cgcgccgggt tttggcgcct cccgcgggcg cccccctcct
4741 cacggcgagc gctgccacgt cagacgaagg gcgcagcgag cgtcctgatc cttccgcccg
4801 gacgctcagg acagcggccc gctgctcata agactcggcc ttagaacccc agtatcagca
4861 gaaggacatt ttaggacggg acttgggtga ctctagggca ctggttttct ttccagagag
4921 cggaacaggc gaggaaaagt agtcccttct cggcgattct gcggagggat ctccgtgggg
4981 cggtgaacgc cgatgattat ataaggacgc gccgggtgtg gcacagctag ttccgtcgca
5041 gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag
5101 cgggctgctg ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg
5161 tgtggagaga ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg
5221 ggttgggggg agcgcagcaa aatggcggct gttcccgagt cttgaatgga agacgcttgt
5281 gaggcgggct gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc aagaacccaa
5341 ggtcttgagg ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg gctggggcac
5401 catctgggga ccctgacgtg aagtttgtca ctgactggag aactcggttt gtcgtctgtt
5461 gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc
5521 gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta taatgcaggg tggggccacc
5581 tgccggtagg tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg ggcctagggt
5641 aggctctcct gaatcgacag gcgccggacc tctggtgagg ggagggataa gtgaggcgtc
5701 agtttctttg gtcggtttta tgtacctatc ttcttaagta gctgaagctc cggttttgaa
5761 ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt
5821 aatcatttgg gtcaatatgt aattttcagt gttagactag taaattgtcc gctaaattct
5881 ggccgttttt ggcttttttg ttagacgaag cttgggctgc aggtcgactc tagaggatcc
5941 agttaacctc gagggcgcgc ccatggacaa gaagtattct atcggactgg ccatcgggac
6001 taatagcgtc gggtgggccg tgatcactga cgagtacaag gtgccctcta agaagttcaa
6061 ggtgctcggg aacaccgacc ggcattccat caagaaaaat ctgatcggag ctctcctctt
6121 tgattcaggg gagaccgctg aagcaacccg cctcaagcgg actgctagac ggcggtacac
6181 caggaggaag aaccggattt gttaccttca agagatattc tccaacgaaa tggcaaaggt
6241 cgacgacagc ttcttccata ggctggaaga atcattcctc gtggaagagg ataagaagca
6301 tgaacggcat cccatcttcg gtaatatcgt cgacgaggtg gcctatcacg agaaataccc
6361 aaccatctac catcttcgca aaaagctggt ggactcaacc gacaaggcag acctccggct
6421 tatctacctg gccctggccc acatgatcaa gttcagaggc cacttcctga tcgagggcga
6481 cctcaatcct gacaatagcg atgtggataa actgttcatc cagctggtgc agacttacaa
6541 ccagctcttt gaagagaacc ccatcaatgc aagcggagtc gatgccaagg ccattctgtc
6601 agcccggctg tcaaagagcc gcagacttga gaatcttatc gctcagctgc cgggtgaaaa
6661 gaaaaatgga ctgttcggga acctgattgc tctttcactt gggctgactc ccaatttcaa
6721 gtctaatttc gacctggcag aggatgccaa gctgcaactg tccaaggaca cctatgatga
6781 cgatctcgac aacctcctgg cccagatcgg tgaccaatac gccgaccttt tccttgctgc
6841 taagaatctt tctgacgcca tcctgctgtc tgacattctc cgcgtgaaca ctgaaatcac
6901 caaggcccct ctttcagctt caatgattaa gcggtatgat gagcaccacc aggacctgac
6961 cctgcttaag gcactcgtcc ggcagcagct tccggagaag tacaaggaaa tcttctttga
7021 ccagtcaaag aatggatacg ccggctacat cgacggaggt gcctcccaag aggaatttta
7081 taagtttatc aaacctatcc ttgagaagat ggacggcacc gaagagctcc tcgtgaaact
7141 gaatcgggag gatctgctgc ggaagcagcg cactttcgac aatgggagca ttccccacca
7201 gatccatctt ggggagcttc acgccatcct tcggcgccaa gaggacttct acccctttct
7261 taaggacaac agggagaaga ttgagaaaat tctcactttc cgcatcccct actacgtggg
7321 acccctcgcc agaggaaata gccggtttgc ttggatgacc agaaagtcag aagaaactat
7381 cactccctgg aacttcgaag aggtggtgga caagggagcc agcgctcagt cattcatcga
7441 acggatgact aacttcgata agaacctccc caatgagaag gtcctgccga aacattccct
7501 gctctacgag tactttaccg tgtacaacga gctgaccaag gtgaaatatg tcaccgaagg
7561 gatgaggaag cccgcattcc tgtcaggcga acaaaagaag gcaattgtgg accttctgtt
7621 caagaccaat agaaaggtga ccgtgaagca gctgaaggag gactatttca agaaaattga
7681 atgcttcgac tctgtggaga ttagcggggt cgaagatcgg ttcaacgcaa gcctgggtac
7741 ctaccatgat ctgcttaaga tcatcaagga caaggatttt ctggacaatg aggagaacga
7801 ggacatcctt gaggacattg tcctgactct cactctgttc gaggaccggg aaatgatcga
7861 ggagaggctt aagacctacg cccatctgtt cgacgataaa gtgatgaagc aacttaaacg
7921 gagaagatat accggatggg gacgccttag ccgcaaactc atcaacggaa tccgggacaa
7981 acagagcgga aagaccattc ttgatttcct taaaagcgac ggattcgcta atcgcaactt
8041 catgcaactt atccatgatg attccctgac ctttaaggag gacatccaga aggcccaagt
8101 gtctggacaa ggtgactcac tgcacgagca tatcgcaaat ctggctggtt cacccgctat
8161 taagaagggt attctccaga ccgtgaaagt cgtggacgag ctggtcaagg tgatgggtcg
8221 ccataaacca gagaacattg tcatcgagat ggccagggaa aaccagacta cccagaaggg
8281 acagaagaac agcagggagc ggatgaaaag aattgaggaa gggattaagg agctcgggtc
8341 acagatcctt aaagagcacc cggtggaaaa cacccagctt cagaatgaga agctctatct
8401 gtactacctt caaaatggac gcgatatgta tgtggaccaa gagcttgata tcaacaggct
8461 ctcagactac gacgtggacg ccatcgtccc tcagagcttc ctcaaagacg actcaattga
8521 caataaggtg ctgactcgct cagacaagaa ccggggaaag tcagataacg tgccctcaga
8581 ggaagtcgtg aaaaagatga agaactattg gcgccagctt ctgaacgcaa agctgatcac
8641 tcagcggaag ttcgacaatc tcactaaggc tgagaggggc ggactgagcg aactggacaa
8701 agcaggattc attaaacggc aacttgtgga gactcggcag attactaaac atgtcgccca
8761 aatccttgac tcacgcatga ataccaagta cgacgaaaac gacaaactta tccgcgaggt
8821 gaaggtgatt accctgaagt ccaagctggt cagcgatttc agaaaggact ttcaattcta
8881 caaagtgcgg gagatcaata actatcatca tgctcatgac gcatatctga atgccgtggt
8941 gggaaccgcc ctgatcaaga agtacccaaa gctggaaagc gagttcgtgt acggagacta
9001 caaggtctac gacgtgcgca agatgattgc caaatctgag caggagatcg gaaaggccac
9061 cgcaaagtac ttcttctaca gcaacatcat gaatttcttc aagaccgaaa tcacccttgc
9121 aaacggtgag atccggaaga ggccgctcat cgagactaat ggggagactg gcgaaatcgt
9181 gtgggacaag ggcagagatt tcgctaccgt gcgcaaagtg ctttctatgc ctcaagtgaa
9241 catcgtgaag aaaaccgagg tgcaaaccgg aggcttttct aaggaatcaa tcctccccaa
9301 gcgcaactcc gacaagctca ttgcaaggaa gaaggattgg gaccctaaga agtacggcgg
9361 attcgattca ccaactgtgg cttattctgt cctggtcgtg gctaaggtgg aaaaaggaaa
9421 gtctaagaag ctcaagagcg tgaaggaact gctgggtatc accattatgg agcgcagctc
9481 cttcgagaag aacccaattg actttctcga agccaaaggt tacaaggaag tcaagaagga
9541 ccttatcatc aagctcccaa agtatagcct gttcgaactg gagaatgggc ggaagcggat
9601 gctcgcctcc gctggcgaac ttcagaaggg taatgagctg gctctcccct ccaagtacgt
9661 gaatttcctc taccttgcaa gccattacga gaagctgaag gggagccccg aggacaacga
9721 gcaaaagcaa ctgtttgtgg agcagcataa gcattatctg gacgagatca ttgagcagat
9781 ttccgagttt tctaaacgcg tcattctcgc tgatgccaac ctcgataaag tccttagcgc
9841 atacaataag cacagagaca aaccaattcg ggagcaggct gagaatatca tccacctgtt
9901 caccctcacc aatcttggtg cccctgccgc attcaagtac ttcgacacca ccatcgaccg
9961 gaaacgctat acctccacca aagaagtgct ggacgccacc ctcatccacc agagcatcac
10021 cggactttac gaaactcgga ttgacctctc acagctcgga ggggatggtg gcggaggctc
10081 gccaaaaaag aagagaaagg tagacccaaa gaaaaaacga aaagtagatc cgaaaaagaa
10141 gaggaaggtg ggatccctgc ccacctgcag ctgtcttgat cgagttatac aaaaagacaa
10201 aggcccatat tatacacacc ttggggcagg accaagtgtt gctgctgtca gggaaatcat
10261 ggagaatagg tatggtcaaa aaggaaacgc aataaggata gaaatagtag tgtacaccgg
10321 taaagaaggg aaaagctctc atgggtgtcc aattgctaag tgggttttaa gaagaagcag
10381 tgatgaagaa aaagttcttt gtttggtccg gcagcgtaca ggccaccact gtccaactgc
10441 tgtgatggtg gtgctcatca tggtgtggga tggcatccct cttccaatgg ccgaccggct
10501 atacacagag ctcacagaga atctaaagtc atacaatggg caccctaccg acagaagatg
10561 caccctcaat gaaaatcgta cctgtacatg tcaaggaatt gatccagaga cttgtggagc
10621 ttcattctct tttggctgtt catggagtat gtactttaat ggctgtaagt ttggtagaag
10681 cccaagcccc agaagattta gaattgatcc aagctctccc ttacatgaaa aaaaccttga
10741 agataactta cagagtttgg ctacacgatt agctccaatt tataagcagt atgctccagt
10801 agcttaccaa aatcaggtgg aatatgaaaa tgttgcccga gaatgtcggc ttggcagcaa
10861 ggaaggtcga cccttctctg gggtcactgc ttgcctggac ttctgtgctc atccccacag
10921 ggacattcac aacatgaata atggaagcac tgtggtttgt accttaactc gagaagataa
10981 ccgctctttg ggtgttattc ctcaagatga gcagctccat gtgctacctc tttataagct
11041 ttcagacaca gatgagtttg gctccaagga aggaatggaa gccaagatca aatctggggc
11101 catcgaggtc ctggcacccc gccgcaaaaa aagaacgtgt ttcactcagc ctgttccccg
11161 ttctggaaag aagagggctg cgatgatgac agaggttctt gcacataaga taagggcagt
11221 ggaaaagaaa cctattcccc gaatcaagcg gaagaataac tcaacaacaa caaacaacag
11281 taagccttcg tcactgccaa ccttagggag taacactgag accgtgcaac ctgaagtaaa
11341 aagtgaaacc gaaccccatt ttatcttaaa aagttcagac aacactaaaa cttattcgct
11401 gatgccatcc gctcctcacc cagtgaaaga ggcatctcca ggcttctcct ggtccccgaa
11461 gactgcttca gccacaccag ctccactgaa gaatgacgca acagcctcat gcgggttttc
11521 agaaagaagc agcactcccc actgtacgat gccttcggga agactcagtg gtgccaatgc
11581 tgcagctgct gatggccctg gcatttcaca gcttggcgaa gtggctcctc tccccaccct
11641 gtctgctcct gtgatggagc ccctcattaa ttctgagcct tccactggtg tgactgagcc
11701 gctaacgcct catcagccaa accaccagcc ctccttcctc acctctcctc aagaccttgc
11761 ctcttctcca atggaagaag atgagcagca ttctgaagca gatgagcctc catcagacga
11821 acccctatct gatgaccccc tgtcacctgc tgaggagaaa ttgccccaca ttgatgagta
11881 ttggtcagac agtgagcaca tctttttgga tgcaaatatt ggtggggtgg ccatcgcacc
11941 tgctcacggc tcggttttga ttgagtgtgc ccggcgagag ctgcacgcta ccactcctgt
12001 tgagcacccc aaccgtaatc atccaacccg cctctccctt gtcttttacc agcacaaaaa
12061 cctaaataag ccccaacatg gttttgaact aaacaagatt aagtttgagg ctaaagaagc
12121 taagaataag aaaatgaagg cctcagagca aaaagaccag gcagctaatg aaggtccaga
12181 acagtcctct gaagtaaatg aattgaacca aattccttct cataaagcat taacattaac
12241 ccatgacaat gttgtcaccg tgtcccctta tgctctcaca cacgttgcgg ggccctataa
12301 ccattgggtc tgagaattcg atatcaagct tatcgataat caacctctgg attacaaaat
12361 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
12421 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
12481 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
12541 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
12601 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
12661 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
12721 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
12781 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
12841 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
12901 gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgagacc tagaaaaaca
12961 tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc
13021 acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac
13081 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
13141 aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta
13201 cttccctgat tggcagaact acacaccagg gccagggatc agatatccac tgacctttgg
13261 atggtgctac aagctagtac cagttgagca agagaaggta gaagaagcca atgaaggaga
13321 gaacacccgc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt
13381 attagagtgg aggtttgaca gccgcctagc atttcatcac atggcccgag agctgcatcc
13441 ggactgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
13501 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
13561 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
13621 aatctctagc agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
13681 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
13741 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
13801 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
13861 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
13921 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
13981 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
14041 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
14101 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
14161 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
14221 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
14281 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
14341 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
14401 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
14461 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
14521 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
14581 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
14641 gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca
14701 tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc
14761 agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac
14821 cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac
14881 gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg
14941 gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac
15001 ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag
15061 ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga
15121 cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
15181 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
15241 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
15301 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
15361 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
15421 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
15481 ggaagcataa agtgtaaagc c
//
