| 质粒名称: | lentiSAM v2 (Puro) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 92062 |
| 存储实验室: | Adam Karpf |
|
载体骨架基本信息 |
|
| 载体名称: | lentiSAM v2 |
| 载体出品公司: | Feng Zhang lab |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 13713 bp |
| 完整质粒大小: | -- |
| 载体修饰: | 采用Bsp1407I和EcoRI消化法去除T2A稻瘟病素。P2A嘌呤霉素通过PCR扩增的方法加入到这些限制性位点。 |
| 载体类型: | 哺乳动物表达，慢病毒，CRISPR |
| 筛选标记: | 嘌呤霉素 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | U6 (sgRNA) and EF1a (dCas9-VP64) |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 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: | lentiSAM v2 (Puro) was a gift from Adam Karpf (Addgene plasmid # 92062 ; http://n2t.net/addgene:92062 ; RRID:Addgene_92062) |
买家导航
lentiSAM v2 (Puro)载体质粒基本信息
lentiSAM v2 (Puro)质粒图谱载体图谱和lentiSAM v2 (Puro)载体序列质粒序列多克隆位点信息
质粒图谱.png)
lentiSAM v2 (Puro)质粒载体简介
lentiSAM v2 (Puro)质粒序列载体序列
LOCUS lentiSAM v2 (Puro) 13713 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Modified version of lentiSAM v2, a lenti sgRNA cloning backbone with
MS2 loops at tetraloop/stemloop 2, dCas9-VP64, and puro resistance
marker. Contains BsmBI sites for insertion of spacer sequences..
ACCESSION .
VERSION .
KEYWORDS lentiSAM v2 (Puro)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13713)
TITLE Lenti CRISPR Activate
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 13713)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..13713
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(84..104)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 89..418
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 269..404
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 331..350
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 466..513
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 532..906
/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 1036..1157
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1073..1092)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1127..1146
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(1206..1222)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1206..1222)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1219..1241)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1230..1246
/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(1254..1284)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1299..1320
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1437..1454)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1608..2196)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1688..1707)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2367..3227)
/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 2990..3009
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3228..3332)
/gene="bla"
/label=AmpR promoter
primer_bind complement(3407..3426)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 3598..3977
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 3979..4177
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 4132..4152
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 4195..4375
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 4422..4547
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 5040..5273
/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 5800..5917
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 5967..6207
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 5967..5987
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 6138..6157
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 6253..6271
/label=MS2 stem loop
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 6323..6341
/label=MS2 stem loop
/note="stem loop that binds the bacteriophage MS2 coat
protein"
promoter 6428..6639
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
CDS 6656..6703
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 6704..10804
/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 10859..10879
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 10904..11053
/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 11075..11131
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS 11132..11728
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind 11625..11645
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 11765..12353
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(11818..11838)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(12236..12247)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(12356..12372)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(12357..12373)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 12878..13058
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
primer_bind complement(13084..13101)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 13090..13314
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 13360..75
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(13447..13466)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 13657..13678
/label=F1ori-F
/note="F1 origin, forward primer"
ORIGIN
1 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
61 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
121 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
181 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
241 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
301 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
361 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
421 gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga caattaatca
481 tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac catggccaag
541 ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg
601 accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg
661 gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg
721 gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc
781 acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg
841 cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag
901 gactgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
961 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
1021 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
1081 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
1141 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
1201 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
1261 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
1321 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
1381 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
1441 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
1501 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
1561 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
1621 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
1681 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
1741 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
1801 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
1861 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
1921 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
1981 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
2041 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
2101 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
2161 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
2221 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
2281 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
2341 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
2401 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
2461 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
2521 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
2581 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
2641 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
2701 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
2761 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
2821 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
2881 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
2941 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
3001 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
3061 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
3121 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
3181 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
3241 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
3301 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
3361 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
3421 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
3481 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
3541 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
3601 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
3661 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
3721 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
3781 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
3841 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
3901 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
3961 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
4021 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
4081 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
4141 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
4201 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
4261 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
4321 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
4381 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
4441 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
4501 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
4561 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
4621 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
4681 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
4741 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
4801 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
4861 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
4921 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
4981 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
5041 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
5101 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
5161 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
5221 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
5281 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
5341 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
5401 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
5461 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
5521 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
5581 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
5641 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
5701 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
5761 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
5821 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
5881 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
5941 agatccagtt tggttaatta gctagcgagg gcctatttcc catgattcct tcatatttgc
6001 atatacgata caaggctgtt agagagataa ttggaattaa tttgactgta aacacaaaga
6061 tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa
6121 aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct
6181 tggctttata tatcttgtgg aaaggacgaa acaccggaga cgggataccg tctctgtttt
6241 agagctaggc caacatgagg atcacccatg tctgcagggc ctagcaagtt aaaataaggc
6301 tagtccgtta tcaacttggc caacatgagg atcacccatg tctgcagggc caagtggcac
6361 cgagtcggtg ctttttttgg atcctaggtc ttgaaaggag tgggaattgg ctccggtgcc
6421 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
6481 aattgatccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
6541 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
6601 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg cgtacggcca ccatgaaaag
6661 gccggcggcc acgaaaaagg ccggccaggc aaaaaagaaa aaggacaaga agtacagcat
6721 cggcctggcc atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt
6781 gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct
6841 gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac
6901 cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag
6961 caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt
7021 ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc
7081 ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga
7141 caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca
7201 cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca
7261 gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga
7321 cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc
7381 ccagctgccc ggcgagaaga agaatggcct gttcggcaac ctgattgccc tgagcctggg
7441 cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag
7501 caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc
7561 cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag
7621 agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga
7681 gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta
7741 caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg acggcggagc
7801 cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga
7861 ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa
7921 cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga
7981 agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg
8041 catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag
8101 aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgcttc
8161 cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt
8221 gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc tgaccaaagt
8281 gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc
8341 catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga
8401 ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt
8461 caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct
8521 ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga
8581 ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt
8641 gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat
8701 caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg
8761 cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga
8821 catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct
8881 ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct
8941 cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa
9001 ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg
9061 catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca
9121 gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga
9181 actggacatc aaccggctgt ccgactacga tgtggaccac atcgtgcctc agagctttct
9241 gaaggacgac tccatcgaca acaaggtgct gaccagaagc gacaaggccc ggggcaagag
9301 cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc ggcagctgct
9361 gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg agagaggcgg
9421 cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat
9481 cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaatga
9541 caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg
9601 gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc
9661 ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc tggaaagcga
9721 gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca
9781 ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa
9841 gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg agacaaacgg
9901 cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc ggaaagtgct
9961 gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa
10021 agagtctatc ctgcccaaga ggaacagcga taagctgatc gccagaaaga aggactggga
10081 ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc tggtggtggc
10141 caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac
10201 catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta
10261 caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga
10321 aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa acgaactggc
10381 cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg
10441 ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc actacctgga
10501 cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct
10561 ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag agcaggccga
10621 gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct tcaagtactt
10681 tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg acgccaccct
10741 gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg
10801 cgacagcgct ggaggaggtg gaagcggagg aggaggaagc ggaggaggag gtagcggacc
10861 taagaaaaag aggaaggtgg cggccgctgg atccggacgg gctgacgcat tggacgattt
10921 tgatctggat atgctgggaa gtgacgccct cgatgatttt gaccttgaca tgcttggttc
10981 ggatgccctt gatgactttg acctcgacat gctcggcagt gacgcccttg atgatttcga
11041 cctggacatg ctgattaact gtacaggcag tggagcaaca aacttctctc tgctgaaaca
11101 agccggagat gtcgaagaga atcctggacc gaccgagtac aagcccacgg tgcgcctcgc
11161 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
11221 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
11281 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
11341 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
11401 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
11461 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggagtctc
11521 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
11581 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
11641 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
11701 gtgcatgacc cgcaagcccg gtgcctgaac gcgttaagtc gaattcgata tcaagcttat
11761 cggtaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
11821 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
11881 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
11941 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
12001 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
12061 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
12121 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
12181 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
12241 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
12301 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata
12361 ccgtcgacct cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac
12421 caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt ttccagtcac
12481 acctcaggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt
12541 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga
12601 tctgtggatc taccacacac aaggctactt ccctgattgg cagaactaca caccagggcc
12661 agggatcaga tatccactga cctttggatg gtgctacaag ctagtaccag ttgagcaaga
12721 gaaggtagaa gaagccaatg aaggagagaa cacccgcttg ttacaccctg tgagcctgca
12781 tgggatggat gacccggaga gagaagtatt agagtggagg tttgacagcc gcctagcatt
12841 tcatcacatg gcccgagagc tgcatccgga ctgtactggg tctctctggt tagaccagat
12901 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
12961 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
13021 cctcagaccc ttttagtcag tgtggaaaat ctctagcagg gcccgtttaa acccgctgat
13081 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
13141 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
13201 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
13261 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
13321 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
13381 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
13441 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
13501 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
13561 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
13621 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
13681 caacactcaa ccctatctcg gtctattctt ttg
//
