| 质粒名称: | lentiCRISPR v2 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 52961 |
| 存储实验室: | Feng Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | Custom |
| 载体抗性: | 氨苄霉素 |
| 空载体大小: | 10000 bp |
| 完整质粒大小: | 14873 bp |
| 载体类型: | 哺乳细胞表达，Lentiviral, 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: | Cas9 |
| 物种来源: | Synthetic |
| 插入大小: | 4200 bp |
| 基因又名: | S. pyogenes CRISPR-Cas9 |
| 启动子 | EFS-NS |
| 标签/融合蛋白: | FLAG (C terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI, XbaI, AfeI(未破坏) |
| 3' 克隆位点: | BamHI(未破坏) |
| 5' 测序引物 | hEF1aprom-F |
| 3' 测序引物 | WPRE-R |
|
文献引用方法 |
|
| 材料和方法部分: | lentiCRISPR v2 was a gift from Feng Zhang (爱迪基因 plasmid # 52961) |
| 参考文献部分: | Improved vectors and genome-wide libraries for CRISPR screening. Sanjana NE, Shalem O, Zhang F. Nat Methods. 2014 Aug;11(8):783-4. doi: 10.1038/nmeth.3047. 10.1038/nmeth.3047 PubMed 25075903 |
lentiCRISPR v2
价格：1000元
联系方式：I47-825O-882O
买家导航
质粒基本信息
质粒图谱信息
质粒简介
质粒序列
LOCUS lentiCRISPR v2 14873 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION Replaces original lentiCRISPRv1 (Addgene Plasmid 49535) and produces
~10-fold higher titer virus. 3rd generation lentiviral backbone..
ACCESSION .
VERSION .
KEYWORDS lentiCRISPR v2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14873)
AUTHORS Sanjana NE, Shalem O, Zhang F
TITLE Improved vectors and genome-wide libraries for CRISPR screening.
JOURNAL Nat Methods. 2014 Aug;11(8):783-4. doi: 10.1038/nmeth.3047.
PUBMED 25075903
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|lentiCRISPR v2|
FEATURES Location/Qualifiers
source 1..14873
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..199
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 154..174
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 217..397
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 444..569
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1062..1295
/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 1822..1939
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 1990..2230
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 1990..2010
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 2161..2180
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 4120..4195
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter 4257..4468
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
regulatory 4487..4496
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 4493..8596
/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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
CDS 8597..8644
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 8645..8668
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 8678..8734
/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 8735..9331
/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 9228..9248
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 9347..9935
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(9400..9420)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(9818..9829)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 10007..10240
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
primer_bind complement(10266..10283)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 10272..10496
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 10542..10970
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(10629..10648)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 10839..10860
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(10979..10999)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 10984..11313
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 11164..11299
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 11226..11245
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 11361..11408
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 11427..11801
/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 11931..12052
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(11968..11987)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 12022..12041
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(12101..12117)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(12101..12117)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(12114..12136)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 12125..12141
/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(12149..12179)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 12194..12215
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(12332..12349)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(12503..13091)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(12583..12602)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(13262..14122)
/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 13885..13904
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(14123..14227)
/gene="bla"
/label=AmpR promoter
primer_bind complement(14302..14321)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 14493..14872
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
ORIGIN
1 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
61 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
121 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
181 gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc
241 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
301 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
361 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
421 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
481 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
541 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
601 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
661 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
721 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
781 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
841 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
901 agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
961 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
1021 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
1081 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
1141 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
1201 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
1261 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
1321 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
1381 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
1441 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
1501 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
1561 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
1621 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
1681 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
1741 agagacagat ccattcgatt agtgaacgga tcggcactgc gtgcgccaat tctgcagaca
1801 aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg
1861 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat
1921 tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag tttggttaat
1981 taaggtaccg agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct
2041 gttagagaga taattagaat taatttgact gtaaacacaa agatattagt acaaaatacg
2101 tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg
2161 gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg
2221 tggaaaggac gaaacaccgg agacggttgt aaatgagcac acaaaataca catgctaaaa
2281 tattatattc tatgaccttt ataaaatcaa ccaaaatctt ctttttaata actttagtat
2341 caataattag aatttttatg ttcctttttg caaactttta ataaaaatga gcaaaataaa
2401 aaaacgctag ttttagtaac tcgcgttgtt ttcttcacct ttaataatag ctactccacc
2461 acttgttcct aagcggtcag ctcctgcttc aatcattttt tgagcatctt caaatgttct
2521 aactccacca gctgctttaa ctaaagcatt gtctttaaca actgacttca ttagtttaac
2581 atcttcaaat gttgcacctg attttgaaaa tcctgttgat gttttaacaa attctaatcc
2641 agcttcaaca gctatttcac aagctttcat gatttcttct tttgttaata aacaattttc
2701 cataatacat ttaacaacat gtgatccagc tgcttttttt acagctttca tgtcttctaa
2761 aactaattca taatttttgt cttttaatgc accaatattt aataccatat caatttctgt
2821 tgcaccatct ttaattgctt cagaaacttc gaatgctttt gtagctgttg tgcatgcacc
2881 tagaggaaaa cctacaacat ttgttattcc tacatttgtg ccttttaata attctttaca
2941 atagcttgtt caatatgaat taacacaaac tgttgcaaaa tcaaattcaa ttgcttcatc
3001 acataattgt ttaatttcag ctttcgtagc atcttgtttt aataatgtgt gatctatata
3061 tttgtttagt ttcatttttt ctcctatata ttcattttta attttaattc tttaataatt
3121 tcgtctactt taactttagc gttttgaaca gattcaccaa cacctataaa ataaattttt
3181 agtttaggtt cagttccact tgggcgaaca gcaaatcatg acttatcttc taaataaaat
3241 tttagtaagt cttgtcctgg catattatac attccatcga tgtagtcttc aacattaaca
3301 actttaagtc cagcaatttg agttaagggt gttgctctca atgatttcat taatggttca
3361 atttttaatt tcttttcttc tggtttaaaa ttcaagttta aagtgaaagt gtaatatgca
3421 cccatttctt taaataaatc ttctaaatag tctactaatg ttttattttg ttttttataa
3481 aatcaagcag cctctgctat taatatagaa gcttgtattc catctttatc tctagctgag
3541 tcatcaatta catatccata actttcttca taagcaaaaa caaaatttaa tccgttatct
3601 tcttctttag caatttctct acccattcat ttaaatccag ttaaagtttt tacaatatta
3661 actccatatt tttcatgagc gattctatca cccaaatcac ttgttacaaa acttgaatat
3721 agagccggat tttttggaat gctatttaag cgttttagat ttgataattt tcaatcaatt
3781 aaaattggtc ctgtttgatt tccatctaat cttacaaaat gaccatcatg ttttattgcc
3841 attccaaatc tgtcagcatc tgggtcattc ataataataa tatctgcatc atgtttaata
3901 ccatattcaa gcggtatttt tcatgcagga tcaaattctg gatttggatt tacaacattt
3961 ttaaatgttt catcttcaaa tgcatgctct tcaacctcaa taacgttata tcctgattca
4021 cgtaatattt ttggggtaaa tttagttcct gttccattaa ctgcgctaaa aataattttt
4081 aaatcttttt tagcttcttg ctcttttttg tacgtctctg ttttagagct agaaatagca
4141 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
4201 tgaattcgct agctaggtct tgaaaggagt gggaattggc tccggtgccc gtcagtgggc
4261 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgatccgg
4321 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
4381 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
4441 tcgcaacggg tttgccgcca gaacacagga ccggttctag agcgctgcca ccatggacaa
4501 gaagtacagc atcggcctgg acatcggcac caactctgtg ggctgggccg tgatcaccga
4561 cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc ggcacagcat
4621 caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg aggccacccg
4681 gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct gctatctgca
4741 agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca gactggaaga
4801 gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg gcaacatcgt
4861 ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa agaaactggt
4921 ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc acatgatcaa
4981 gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa
5041 gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc ccatcaacgc
5101 cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca gacggctgga
5161 aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa acctgattgc
5221 cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa
5281 actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg
5341 cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag
5401 cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa
5461 gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct
5521 gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat
5581 tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat
5641 ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg
5701 gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct
5761 gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat
5821 cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc
5881 ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga
5941 caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc
6001 caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtataacga
6061 gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga
6121 gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca
6181 gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt
6241 ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa ttatcaagga
6301 caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg tgctgaccct
6361 gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg cccacctgtt
6421 cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg gcaggctgag
6481 ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc tggatttcct
6541 gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg acagcctgac
6601 ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc tgcacgagca
6661 cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt
6721 ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat
6781 ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg
6841 gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa
6901 cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta
6961 cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacc atatcgtgcc
7021 tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa gcgacaagaa
7081 ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg
7141 gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc tgaccaaggc
7201 cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga
7261 aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta
7321 cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt ccaagctggt
7381 gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca
7441 cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa
7501 gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc
7561 caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat
7621 gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat
7681 cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt ttgccaccgt
7741 gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg
7801 cggcttcagc aaagagtcta tcctgcccaa gaggaacagc gataagctga tcgccagaaa
7861 gaaggactgg gaccctaaga agtacggcgg cttcgacagc cccaccgtgg cctattctgt
7921 gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct
7981 gctggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga
8041 agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct
8101 gttcgagctg gaaaacggcc ggaagagaat gctggcctct gccggcgaac tgcagaaggg
8161 aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga
8221 gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa
8281 gcactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc
8341 cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata agcccatcag
8401 agagcaggcc gagaatatca tccacctgtt taccctgacc aatctgggag cccctgccgc
8461 cttcaagtac tttgacacca ccatcgaccg gaagaggtac accagcacca aagaggtgct
8521 ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga tcgacctgtc
8581 tcagctggga ggcgacaagc gacctgccgc cacaaagaag gctggacagg ctaagaagaa
8641 gaaagattac aaagacgatg acgataaggg atccggcgca acaaacttct ctctgctgaa
8701 acaagccgga gatgtcgaag agaatcctgg accgaccgag tacaagccca cggtgcgcct
8761 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta
8821 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca
8881 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
8941 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
9001 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
9061 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggagt
9121 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
9181 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
9241 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
9301 ctggtgcatg acccgcaagc ccggtgcctg aacgcgttaa gtcgacaatc aacctctgga
9361 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
9421 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
9481 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
9541 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
9601 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
9661 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
9721 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
9781 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
9841 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
9901 gacgagtcgg atctcccttt gggccgcctc cccgcgtcga ctttaagacc aatgacttac
9961 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
10021 cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag
10081 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
10141 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
10201 tccctcagac ccttttagtc agtgtggaaa atctctagca gggcccgttt aaacccgctg
10261 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
10321 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
10381 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
10441 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
10501 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
10561 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
10621 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
10681 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
10741 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
10801 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
10861 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
10921 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
10981 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
11041 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
11101 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
11161 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
11221 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
11281 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
11341 ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat
11401 aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt ccggtgctca
11461 ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg
11521 acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg
11581 cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg
11641 acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc
11701 cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg
11761 ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg
11821 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
11881 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
11941 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
12001 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
12061 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
12121 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
12181 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
12241 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
12301 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
12361 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
12421 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
12481 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
12541 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
12601 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
12661 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
12721 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
12781 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
12841 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
12901 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
12961 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
13021 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
13081 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
13141 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
13201 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
13261 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
13321 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
13381 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
13441 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
13501 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
13561 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
13621 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
13681 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
13741 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
13801 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
13861 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
13921 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
13981 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
14041 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
14101 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
14161 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
14221 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
14281 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
14341 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
14401 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
14461 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
14521 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
14581 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
14641 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
14701 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
14761 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
14821 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tgg
//
