| 质粒名称: | pXPR_120 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 96917 |
| 存储实验室: | John Doench, David Root |
|
载体骨架基本信息 |
|
| 载体名称: | pXPR |
| 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: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，慢病毒 |
| 筛选标记: | 杀螨素 |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | dCas9 |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | D10A, N863A |
| GenBank编号 | -- |
| 启动子 | EF1a |
| 标签/融合蛋白: | VPR（插入件上的C端子） |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pXPR_120 was a gift from John Doench & David Root (Addgene plasmid # 96917 ; http://n2t.net/addgene:96917 ; RRID:Addgene_96917) |
| 参考文献部分: | Orthologous CRISPR-Cas9 enzymes for combinatorial genetic screens. Najm FJ, Strand C, Donovan KF, Hegde M, Sanson KR, Vaimberg EW, Sullender ME, Hartenian E, Kalani Z, Fusi N, Listgarten J, Younger ST, Bernstein BE, Root DE, Doench JG. Nat Biotechnol. 2018 Feb;36(2):179-189. doi: 10.1038/nbt.4048. Epub 2017 Dec 18. 10.1038/nbt.4048 PubMed 29251726 |
买家导航
pXPR120载体基本信息
pXPR120载体图谱质粒图谱和多克隆位点信息
pXPR120载体简介
pXPR120载体序列
LOCUS pXPR_120 15513 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION for CRISPRa, lentiviral expression of dCas9-VPR 2A BlastR.
ACCESSION .
VERSION .
KEYWORDS pXPR_120
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 15513)
AUTHORS Najm FJ, Strand C, Donovan KF, Hegde M, Sanson KR, Vaimberg EW,
Sullender ME, Hartenian E, Kalani Z, Fusi N, Listgarten J, Younger
ST, Bernstein BE, Root DE, Doench JG
TITLE Orthologous CRISPR-Cas9 enzymes for combinatorial genetic screens.
JOURNAL Nat Biotechnol. 2018 Feb;36(2):179-189. doi: 10.1038/nbt.4048. Epub
2017 Dec 18.
PUBMED 29251726
REFERENCE 2 (bases 1 to 15513)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..15513
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(30..49)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind 276..296
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin complement(305..733)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(415..436)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 627..646
/label=F1ori-R
/note="F1 origin, reverse primer"
polyA_signal 779..1003
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind 992..1009
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
LTR 1035..1215
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
primer_bind 1720..1736
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind 1721..1737
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 1740..2328
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS 1846..1857
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind 2255..2275
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2353..2751)
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/label=BSD
/note="confers resistance to blasticidin"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
CDS complement(2752..2805)
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS complement(2821..3390)
/codon_start=1
/product="transcriptional activation domain from the human
herpesvirus 4 (Epstein-Barr virus) replication and
transcription activator Rta/BRLF1 (Hardwick et al., 1992;
Chavez et al., 2015)"
/label=Rta AD
/translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
DTFLNDECLLHAMHISTGLSIFDTSLF"
CDS complement(3409..3765)
/codon_start=1
/product="transcriptional activation domain of human RelA,
also known as p65 (O'Shea and Perkins, 2008)"
/label=RelA (p65) AD
/translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
EDFSSIADMDFSALL"
CDS complement(4195..4215)
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS complement(4240..4389)
/codon_start=1
/product="tetrameric repeat of the minimal activation
domain of herpes simplex virus VP16 (Beerli et al., 1998)"
/label=VP64
/translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
DLDML"
CDS complement(4429..4449)
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS complement(4504..8604)
/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 N863A mutation in the HNH
catalytic domain"
/translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKARGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS complement(8605..8652)
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
promoter complement(8668..9846)
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 8677..9615
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind complement(8700..8720)
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
misc_feature 9976..10093
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
misc_feature 10620..10853
/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 11346..11471
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
LTR 11518..11698
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
promoter complement(11716..11914)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(11741..11761)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 11916..12295
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
primer_bind 12467..12486
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 12561..12665
/gene="bla"
/label=AmpR promoter
CDS 12666..13526
/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(12884..12903)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 13697..14285
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 14186..14205
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 14439..14456
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 14573..14594
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 14609..14639
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 14647..14663
/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)."
primer_bind 14652..14674
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 14671..14687
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 14671..14687
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
polyA_signal 14736..14857
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(14747..14766)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 14801..14820
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
CDS complement(14987..15361)
/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"
promoter complement(15380..15427)
/label=EM7 promoter
/note="synthetic bacterial promoter "
promoter complement(15475..291)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 15489..111
/label=SV40 ori
/note="SV40 origin of replication"
ORIGIN
1 cttctggaat agctcagagg cagaggcggc ctcggcctct gcataaataa aaaaaattag
61 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
121 taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
181 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
241 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
301 aattaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
361 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
421 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
481 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
541 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
601 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
661 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
721 ctacagggcg cgtggggata ccccctagag ccccagctgg ttctttccgc ctcagaagcc
781 atagagccca ccgcatcccc agcatgcctg ctattgtctt cccaatcctc ccccttgctg
841 tcctgcccca ccccaccccc cagaatagaa tgacacctac tcagacaatg cgatgcaatt
901 tcctcatttt attaggaaag gacagtggga gtggcacctt ccagggtcaa ggaaggcacg
961 ggggaggggc aaacaacaga tggctggcaa ctagaaggca cagtcgaggc tgatcagcgg
1021 gtttaaacgg gccctgctag agattttcca cactgactaa aagggtctga gggatctcta
1081 gttaccagag tcacacaaca gacgggcaca cactacttga agcactcaag gcaagcttta
1141 ttgaggctta agcagtgggt tccctagtta gccagagagc tcccaggctc agatctggtc
1201 taaccagaga gacccagtac agtccggatg cagctctcgg gccatgtgat gaaatgctag
1261 gcggctgtca aacctccact ctaatacttc tctctccggg tcatccatcc catgcaggct
1321 cacagggtgt aacaagcggg tgttctctcc ttcattggct tcttctacct tctcttgctc
1381 aactggtact agcttgtagc accatccaaa ggtcagtgga tatctgatcc ctggccctgg
1441 tgtgtagttc tgccaatcag ggaagtagcc ttgtgtgtgg tagatccaca gatcaaggat
1501 atcttgtctt cgttgggagt gaattagccc ttccagtccc cccttttctt ttaaaaagtg
1561 gctaagatct acagctgcct tgtaagtcat tggtcttaaa ggtacctgag gtgtgactgg
1621 aaaacccacc tcctcctcct cttgtgcttc tagccaggca caatcagcat tggtagctgc
1681 tgtattgcta cttgtgattg ctccatgttt ttctaggtct cgaggtcgac ggtatcgatg
1741 cggggaggcg gcccaaaggg agatccgact cgtctgaggg cgaaggcgaa gacgcggaag
1801 aggccgcaga gccggcagca ggccgcggga aggaaggtcc gctggattga gggccgaagg
1861 gacgtagcag aaggacgtcc cgcgcagaat ccaggtggca acacaggcga gcagccaagg
1921 aaaggacgat gatttccccg acaacaccac ggaattgtca gtgcccaaca gccgagcccc
1981 tgtccagcag cgggcaaggc aggcggcgat gagttccgcc gtggcaatag ggagggggaa
2041 agcgaaagtc ccggaaagga gctgacaggt ggtggcaatg ccccaaccag tgggggttgc
2101 gtcagcaaac acagtgcaca ccacgccacg ttgcctgaca acgggccaca actcctcata
2161 aagagacagc aaccaggatt tatacaagga ggagaaaatg aaagccatac gggaagcaat
2221 agcatgatac aaaggcatta aagcagcgta tccacatagc gtaaaaggag caacatagtt
2281 aagaatacca gtcaatcttt cacaaatttt gtaatccaga ggttgattac cgataagctt
2341 gatatcgaat tcttagccct cccacacata accagagggc agcaattcac gaatcccaac
2401 tgccgtcggc tgtccatcac tgtccttcac tatggctttg atcccaggat gcagatcgag
2461 aagcacctgt cggcaccgtc cgcaggggct caagatgccc ctgttctcat ttccgatcgc
2521 gacgatacaa gtcaggttgc cagctgccgc agcagcagca gtgcccagca ccacgagttc
2581 tgcacaaggt cccccagtaa aatgatatac attgacacca gtgaagatgc ggccgtcgct
2641 agagagagct gcgctggcga cgctgtagtc ttcagagatg gggatgctgt tgattgtagc
2701 cgttgctctt tcaatgaggg tggattcttc ttgagacaaa ggcttggcca ttgggccagg
2761 attctcctcg acgtcaccgc atgttagcag acttcctctg ccctctccac tgcctgtaca
2821 gaagaggctg gtgtcgaaga tggacagccc ggtggagatg tgcatggcgt gcagcaggca
2881 ctcgtcgttg aggaaggtgt ccaggatctc gttgagctca ggggtgagcg gcgagtccaa
2941 gttgaggtcc tcggtcatgc tctcgagggt ggtggtgagc tcgtccaggt ggccgcgggg
3001 cggagggtgg gacaggtcca tctggccgca gatggcagcc tcctccttct gggggatgac
3061 ggtgtcggcc atctcgcgca gggccttgac ggcctgggag gtctcctcgt cagggtcctc
3121 cagcaggtgg gaggcctcgg gcgtgacagc aggagcaggg tccagcggct ggggcacagg
3181 cgcaggggtg aggctgccaa ctggctcgtg cacggggcca gttggggttg gtgccaacga
3241 ggcaggcagc gggcggttgg cccagggtga gccaggaggg tggaaggggc ggatccgctt
3301 gggctggcag acctcacgcc cctcgaagac gtccgagatg gcagagccag cctccggctt
3361 gggcaggaac atgccctcgc gggagtcgcg ggagccggag ccagagccca gcagggcaga
3421 gaagtccatg tccgcgatgg aggagaagtc ttcgtcgccg ctcagcaggc cgttgggcag
3481 gcctggagcc cccaggggag caggggcagg gtcaggaggg cgttgggcac cggtcaccag
3541 gcgggtgatg gcctctgggt actccatgag catgggctcg gtggtgtggg gggccacggg
3601 gatgccctgg ttgagcagct gctggaactc ggagttgtcc accgatgcca agtcggtgaa
3661 gacagcaggg tcggtggagt tgcccagcag ggcccccagg tcctcgtcgt cgaactgaag
3721 ttggagcagc gcctcggaca gggtgccctc gcctgcctgg gtaggcttgg gcgcaggagg
3781 cgccacggcc tgcggagggc caggtgccaa cacaggcacc ggggcagggg cctgggccag
3841 ggcagagacc atggcaggag caggagcagg agcaggagcc tggggcagca cctgcggagg
3901 agcaggtgcc aacgcagagg cctgggagat ctggccagag gggaagacca tggtggggaa
3961 ctcgtcgtag ttgatggtgg acagcgagga ggtgaagggg tagggctggg gagcaggctt
4021 gggcacagag gcagaggagc ggctgggcac cgcgatgcgc cgcggaggcg gcctcgggtc
4081 agttgggcca gagaagggcg acttcttcat gatggacttg aaagtttcgt aggtgcgctt
4141 gcgcttttcc tcgatacggt ggcggtcgtc ggtgtcgggc aggtactggc tgccaacttt
4201 gcgcttcttc ttgggagacc cagaggagcg ggagttgatg agcatgtcca ggtcgaagtc
4261 gtcgagggcg tccgagccca gcatgtccag gtcgaagtcg tccagcgcgt cgctgcccag
4321 catgtccagg tcgaagtcgt ccagggcgtc ggagcccagc atgtccaggt cgaagtcgtc
4381 cagggcgtcc gcgcggccgg agccagaggc ctcggatcca gcggccgcca ccttcctctt
4441 tttcttaggt ccgctacctc ctcctccgct tcctcctcct ccgcttccac ctcctccagc
4501 gctgtcgcct cccagctgag acaggtcgat ccgtgtctcg tacaggccgg tgatgctctg
4561 gtggatcagg gtggcgtcca gcacctcttt ggtgctggtg tacctcttcc ggtcgatggt
4621 ggtgtcaaag tacttgaagg cggcaggggc tcccagattg gtcagggtaa acaggtggat
4681 gatattctcg gcctgctctc tgatgggctt atcccggtgc ttgttgtagg cggacagcac
4741 tttgtccaga ttagcgtcgg ccaggatcac tctcttggag aactcgctga tctgctcgat
4801 gatctcgtcc aggtagtgct tgtgctgttc cacaaacagc tgtttctgct cattatcctc
4861 gggggagccc ttcagcttct catagtggct ggccaggtac aggaagttca catatttgga
4921 gggcagggcc agttcgtttc ccttctgcag ttcgccggca gaggccagca ttctcttccg
4981 gccgttttcc agctcgaaca gggagtactt aggcagcttg atgatcaggt cctttttcac
5041 ttctttgtag cccttggctt ccagaaagtc gatgggattc ttctcgaagc tgcttctttc
5101 catgatggtg atccccagca gctctttcac actcttcagt ttcttggact tgcccttttc
5161 cactttggcc accaccagca cagaataggc cacggtgggg ctgtcgaagc cgccgtactt
5221 cttagggtcc cagtccttct ttctggcgat cagcttatcg ctgttcctct tgggcaggat
5281 agactctttg ctgaagccgc ctgtctgcac ctcggtcttt ttcacgatat tcacttgggg
5341 catgctcagc actttccgca cggtggcaaa atcccggccc ttatcccaca cgatctcccc
5401 ggtttcgccg tttgtctcga tcagaggccg cttccggatc tcgccgttgg ccagggtaat
5461 ctcggtcttg aaaaagttca tgatgttgct gtagaagaag tacttggcgg tagccttgcc
5521 gatttcctgc tcgctcttgg cgatcatctt ccgcacgtcg tacaccttgt agtcgccgta
5581 cacgaactcg ctttccagct tagggtactt tttgatcagg gcggttccca cgacggcgtt
5641 caggtaggcg tcgtgggcgt ggtggtagtt gttgatctcg cgcactttgt aaaactggaa
5701 atccttccgg aaatcggaca ccagcttgga cttcagggtg atcactttca cttcccggat
5761 cagcttgtca ttctcgtcgt acttagtgtt catccgggag tccaggatct gtgccacgtg
5821 ctttgtgatc tgccgggttt ccaccagctg tctcttgatg aagccggcct tatccagttc
5881 gctcaggccg cctctctcgg ccttggtcag attgtcgaac tttctctggg taatcagctt
5941 ggcgttcagc agctgccgcc agtagttctt catcttcttc acgacctctt cggagggcac
6001 gttgtcgctc ttgccccggg ccttgtcgct tctggtcagc accttgttgt cgatggagtc
6061 gtccttcaga aagctctgag gcacgatgtg gtccacatcg tagtcggaca gccggttgat
6121 gtccagttcc tggtccacgt acatatcccg cccattctgc aggtagtaca ggtacagctt
6181 ctcgttctgc agctgggtgt tttccacggg gtgttctttc aggatctggc tgcccagctc
6241 tttgatgccc tcttcgatcc gcttcattct ctcgcggctg ttcttctgtc ccttctgggt
6301 ggtctggttc tctctggcca tttcgatcac gatgttctcg ggcttgtgcc ggcccatcac
6361 tttcacgagc tcgtccacca ccttcactgt ctgcaggatg cccttcttaa tggcggggct
6421 gccggccaga ttggcaatgt gctcgtgcag gctatcgccc tggccggaca cctgggcttt
6481 ctggatgtcc tctttaaagg tcaggctgtc gtcgtggatc agctgcatga agtttctgtt
6541 ggcgaagccg tcggacttca ggaaatccag gattgtcttg ccggactgct tgtcccggat
6601 gccgttgatc agcttccggc tcagcctgcc ccagccggtg tatctccgcc gcttcagctg
6661 cttcatcact ttgtcgtcga acaggtgggc ataggttttc agccgttcct cgatcatctc
6721 tctgtcctca aacagtgtca gggtcagcac gatatcttcc agaatgtcct cgttttcctc
6781 attgtccagg aagtccttgt ccttgataat tttcagcaga tcgtggtatg tgcccaggga
6841 ggcgttgaac cgatcttcca cgccggagat ttccacggag tcgaagcact cgattttctt
6901 gaagtagtcc tctttcagct gcttcacggt cactttccgg ttggtcttga acagcaggtc
6961 cacgatggcc tttttctgct cgccgctcag gaaggcgggc tttctcattc cctcggtcac
7021 gtatttcact ttggtcagct cgttatacac ggtgaagtac tcgtacagca ggctgtgctt
7081 gggcagcacc ttctcgttgg gcaggttctt atcgaagttg gtcatccgct cgatgaagct
7141 ctgggcggaa gcgcccttgt ccaccacttc ctcgaagttc cagggggtga tggtttcctc
7201 gctctttctg gtcatccagg cgaatctgct gtttcccctg gccagagggc ccacgtagta
7261 ggggatgcgg aaggtcagga tcttctcgat cttttcccgg ttgtccttca ggaatgggta
7321 aaaatcttcc tgccgccgca gaatggcgtg cagctctccc aggtggatct ggtgggggat
7381 gctgccgttg tcgaaggtcc gctgcttccg cagcaggtcc tctctgttca gcttcacgag
7441 cagttcctcg gtgccgtcca tcttttccag gatgggcttg atgaacttgt agaactcttc
7501 ctggctggct ccgccgtcaa tgtagccggc gtagccgttc ttgctctggt cgaagaaaat
7561 ctctttgtac ttctcaggca gctgctgccg cacgagagct ttcagcaggg tcaggtcctg
7621 gtggtgctcg tcgtatctct tgatcataga ggcgctcagg ggggccttgg tgatctcggt
7681 gttcactctc aggatgtcgc tcagcaggat ggcgtcggac aggttcttgg cggccagaaa
7741 caggtcggcg tactggtcgc cgatctgggc cagcaggttg tccaggtcgt cgtcgtaggt
7801 gtccttgctc agctgcagtt tggcatcctc ggccaggtcg aagttgctct tgaagttggg
7861 ggtcaggccc aggctcaggg caatcaggtt gccgaacagg ccattcttct tctcgccggg
7921 cagctgggcg atcagatttt ccagccgtct gctcttgctc agtctggcag acaggatggc
7981 cttggcgtcc acgccgctgg cgttgatggg gttttcctcg aacagctggt tgtaggtctg
8041 caccagctgg atgaacagct tgtccacgtc gctgttgtcg gggttcaggt cgccctcgat
8101 caggaagtgg ccccggaact tgatcatgtg ggccagggcc agatagatca gccgcaggtc
8161 ggccttgtcg gtgctgtcca ccagtttctt tctcaggtgg tagatggtgg ggtacttctc
8221 gtggtaggcc acctcgtcca cgatgttgcc gaagatgggg tgccgctcgt gcttcttatc
8281 ctcttccacc aggaaggact cttccagtct gtggaagaag ctgtcgtcca ccttggccat
8341 ctcgttgctg aagatctctt gcagatagca gatccggttc ttccgtctgg tgtatcttct
8401 tctggcggtt ctcttcagcc gggtggcctc ggctgtttcg ccgctgtcga acagcagggc
8461 tccgatcagg ttcttcttga tgctgtgccg gtcggtgttg cccagcacct tgaatttctt
8521 gctgggcacc ttgtactcgt cggtgatcac ggcccagccc acagagttgg tgccgatggc
8581 caggccgatg ctgtacttct tgtccttttt cttttttgcc tggccggcct ttttcgtggc
8641 cgccggcctt ttcatggtgg ccgtacgtca cgacacctga aatggaagaa aaaaactttg
8701 aaccactgtc tgaggcttga gaatgaacca agatccaaac tcaaaaaggg caaattccaa
8761 ggagaattac atcaagtgcc aagctggcct aacttcagtc tccacccact cagtgtgggg
8821 aaactccatc gcataaaacc cctcccccca acctaaagac gacgtactcc aaaagctcga
8881 gaactaatcg aggtgcctgg acggcgcccg gtactccgtg gagtcacatg aagcgacggc
8941 tgaggacgga aaggcccttt tcctttgtgt gggtgactca cccgcccgct ctcccgagcg
9001 ccgcgtcctc cattttgagc tccctgcagc agggccggga agcggccatc tttccgctca
9061 cgcaactggt gccgaccggg ccagccttgc cgcccagggc ggggcgatac acggcggcgc
9121 gaggccaggc accagagcag gccggccagc ttgagactac ccccgtccga ttctcggtgg
9181 ccgcgctcgc aggccccgcc tcgccgaaca tgtgcgctgg gacgcacggg ccccgtcgcc
9241 gcccgcggcc ccaaaaaccg aaataccagt gtgcagatct tggcccgcat ttacaagact
9301 atcttgccag aaaaaaagcg tcgcagcagg tcatcaaaaa ttttaaatgg ctagagactt
9361 atcgaaagca gcgagacagg cgcgaaggtg ccaccagatt cgcacgcggc ggccccagcg
9421 cccaggccag gcctcaactc aagcacgagg cgaaggggct ccttaagcgc aaggcctcga
9481 actctcccac ccacttccaa cccgaagctc gggatcaaga atcacgtact gcagccaggt
9541 ggaagtaatt caaggcacgc aagggccata acccgtaaag aggccaggcc cgcgggaacc
9601 acacacggca cttacctgtg ttctggcggc aaacccgttg cgaaaaagaa cgttcacggc
9661 gactactgca cttatatacg gttctccccc accctcggga aaaaggcgga gccagtacac
9721 gacatcactt tcccagttta ccccgcgcca ccttctctag gcaccggttc aattgccgac
9781 ccctcccccc aacttctcgg ggactgtggg cgatgtgcgc tctgcccact gacgggcacc
9841 ggagccaatt cccactcctt tcaagaccta gaaggtccat tagctgcaaa gattcctctc
9901 tgtttaaaac tttatccatc tttgcagcta gctaattaac caaactggat ctctgctgtc
9961 cctgtaataa acccgaaaat tttgaatttt tgtaatttgt ttttgtaatt ctttagtttg
10021 tatgtctgtt gctattatgt ctactattct ttcccctgca ctgtaccccc caatcccccc
10081 ttttctttta aaattgtgga tgaatactgc catttgtctg cagaattggc gcacgcagtg
10141 ccgatccgtt cactaatcga atggatctgt ctctgtctct ctctccacct tcttcttcta
10201 ttccttcggg cctgtcgggt cccctcgggg ttgggaggtg ggtctgaaac gataatggtg
10261 aatatccctg cctaactcta ttcactatag aaagtacagc aaaaactatt cttaaaccta
10321 ccaagcctcc tactatcatt atgaataatt ttatatacca cagccaattt gttatgttaa
10381 accaattcca caaacttgcc catttatcta attccaataa ttcttgttca ttcttttctt
10441 gctggttttg cgattcttca attaaggagt gtattaagct tgtgtaattg ttaatttctc
10501 tgtcccactc catccaggtc gtgtgattcc aaatctgttc cagagattta ttactccaac
10561 tagcattcca aggcacagca gtggtgcaaa tgagttttcc agagcaaccc caaatcccca
10621 ggagctgttg atcctttagg tatctttcca cagccaggat tcttgcctgg agctgcttga
10681 tgccccagac tgtgagttgc aacagatgct gttgcgcctc aatagccctc agcaaattgt
10741 tctgctgctg cactatacca gacaataatt gtctggcctg taccgtcagc gtcattgacg
10801 ctgcgcccat agtgcttcct gctgctccca agaacccaag gaacaaagct cctattccca
10861 ctgctctttt ttctctctgc accactcttc tctttgcctt ggtgggtgct actcctaatg
10921 gttcaatttt tactacttta tatttatata attcacttct ccaattgtcc ctcatatctc
10981 ctcctccagg tctgaagatc agcggccgct tgctgtgcgg tggtcttact tttgttttgc
11041 tcttcctcta tcttgtctaa agcttccttg gtgtctttta tctctatcct ttgatgcaca
11101 caatagaggg ttgctactgt attatataat gatctaagtt cttctgatcc tgtctgaagg
11161 gatggttgta gctgtcccag tatttgtcta cagccttctg atgtttctaa caggccagga
11221 ttaactgcga atcgttctag ctccctgctt gcccatacta tatgttttaa tttatatttt
11281 ttctttcccc ctggccttaa ccgaattttt tcccatcgcg atctaattct cccccgctta
11341 atactgacgc tctcgcaccc atctctctcc ttctagcctc cgctagtcaa aatttttggc
11401 gtactcacca gtcgccgccc ctcgcctctt gccgtgcgcg cttcagcaag ccgagtcctg
11461 cgtcgagaga gctcctctgg tttccctttc gctttcaagt ccctgttcgg gcgccactgc
11521 tagagatttt ccacactgac taaaagggtc tgagggatct ctagttacca gagtcacaca
11581 acagacgggc acacactact tgaagcactc aaggcaagct ttattgaggc ttaagcagtg
11641 ggttccctag ttagccagag agctcccagg ctcagatctg gtctaaccag agagacccag
11701 tacaggcaaa acgcgctgct tatatagacc tcccaccgta cacgcctacc gcccatttgc
11761 gtcaatgggg cggagttgtt acgacatttt ggaaagtccc gttgattttg gtgccaaaac
11821 aaactcccat tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc aaaccgctat
11881 ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg taatagcgat gactaatacg
11941 tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg
12001 ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt
12061 actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct
12121 attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc
12181 ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta
12241 tgaactaatg accccgtaat tgattactat taataactag tcaataatca atgtcaacgc
12301 gtatatctgg cccgtacatc gcgaagcagc gcaaaacgcc taaccctaag cagattcttc
12361 atgcaattgt cggtcaagcc ttgccttgtt gtagcttaaa ttttgctcgc gcactactca
12421 gcgacctcca acacacaagc agggagcaga tactggctta actatgcggc atcagagcag
12481 attgtactga gagtgcacca taggggatcg ggagatctcc cgatccgtcg acgtcaggtg
12541 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
12601 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
12661 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
12721 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
12781 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
12841 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
12901 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
12961 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
13021 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
13081 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
13141 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
13201 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
13261 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
13321 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
13381 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
13441 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
13501 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
13561 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
13621 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
13681 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
13741 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
13801 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
13861 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
13921 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
13981 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
14041 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
14101 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
14161 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
14221 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
14281 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
14341 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
14401 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
14461 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
14521 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
14581 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
14641 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
14701 agctctagct agaggtcgac ggtatacaga catgataaga tacattgatg agtttggaca
14761 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
14821 tttatttgta accattataa gctgcaataa acaagttggg gtgggcgaag aactccagca
14881 tgagatcccc gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca
14941 acctttcata gaaggcggcg gtggaatcga aatctcgtag cacgtgtcag tcctgctcct
15001 cggccacgaa gtgcacgcag ttgccggccg ggtcgcgcag ggcgaactcc cgcccccacg
15061 gctgctcgcc gatctcggtc atggccggcc cggaggcgtc ccggaagttc gtggacacga
15121 cctccgacca ctcggcgtac agctcgtcca ggccgcgcac ccacacccag gccagggtgt
15181 tgtccggcac cacctggtcc tggaccgcgc tgatgaacag ggtcacgtcg tcccggacca
15241 caccggcgaa gtcgtcctcc acgaagtccc gggagaaccc gagccggtcg gtccagaact
15301 cgaccgctcc ggcgacgtcg cgcgcggtga gcaccggaac ggcactggtc aacttggcca
15361 tggtttagtt cctcaccttg tcgtattata ctatgccgat atactatgcc gatgattaat
15421 tgtcaacacg tgctgatcag atccgaaaat ggatatacaa gctcccggga gctttttgca
15481 aaagcctagg cctccaaaaa agcctcctca cta
//
