| 质粒名称: | pR26 GFP Dest |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 74283 |
| 存储实验室: | Ralf Kuehn |
|
载体骨架基本信息 |
|
| 载体名称: | Bluescript |
| 完整质粒大小: | 14707 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | Chloramphenicol and Ampicillin, 25 & 100 μg/mL |
| 生长温度: | 30 ℃ |
| 宿主菌: | ccdB Survival |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Rosa26 5-homology region |
| 物种来源: | M. musculus (mouse) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | I-SceI(未破坏) |
| 3’克隆位点: | XbaI (未破坏) |
| 5’测序引物 | M13 forward |
| 3’测序引物 | M13 reverse |
|
文献引用方法 |
|
| 材料和方法部分: | pR26 GFP Dest was a gift from Ralf Kuehn (Addgene plasmid # 74283 ; http://n2t.net/addgene:74283 ; RRID:Addgene_74283) |
| 参考文献部分: | Efficient generation of Rosa26 knock-in mice using CRISPR/Cas9 in C57BL/6 zygotes. Chu VT, Weber T, Graf R, Sommermann T, Petsch K, Sack U, Volchkov P, Rajewsky K, Kuhn R. BMC Biotechnol. 2016 Jan 16;16(1):4. doi: 10.1186/s12896-016-0234-4. 10.1186/s12896-016-0234-4 PubMed 26772810 |
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 pR26 GFP Dest 14707 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Gene targeting vector for the mouse Rosa26 locus, including a loxP
flanked stop cassette and GFP, for gateway cloning.
ACCESSION .
VERSION .
KEYWORDS pR26 GFP Dest
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14707)
AUTHORS Chu VT, Weber T, Graf R, Sommermann T, Petsch K, Sack U, Volchkov P,
Rajewsky K, Kuhn R
TITLE Efficient generation of Rosa26 knock-in mice using CRISPR/Cas9 in
C57BL/6 zygotes.
JOURNAL BMC Biotechnol. 2016 Jan 16;16(1):4. doi: 10.1186/s12896-016-0234-4.
PUBMED 26772810
REFERENCE 2 (bases 1 to 14707)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..14707
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 5..1087
/label=Rosa26 left arm
/note="left homology arm from intron 1 of the mouse Rosa26
gene (Soriano, 1999)"
misc_feature 1094..1197
/label=Ad SA
/note="adenovirus major late transcript splice acceptor
(Friedrich and Soriano, 1991)"
protein_bind 1208..1241
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
polyA_signal 1248..1296
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
CDS complement(1303..2106)
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(1414..1433)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 2024..2043
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
intron 2920..2985
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 3115..3135
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 3560..3694
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3597..3616)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3651..3670
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
protein_bind 3714..3747
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
protein_bind 3760..3884
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 3909..3939
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 3993..4652
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(4059..4078)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
CDS 4994..5299
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
primer_bind 5178..5197
/label=ccdB-fwd
/note="ccdB gene, forward primer"
protein_bind complement(5340..5464)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
protein_bind 5499..5546
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
misc_feature 5585..6136
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(5729..5746)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 5956..5975
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 6148..6867
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 6148..6864
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(6193..6214)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(6454..6473)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 6801..6822
/label=EGFP-C
/note="EGFP, forward primer"
protein_bind 6892..6939
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
primer_bind complement(6964..6980)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6964..6980)
/label=pBluescriptSK
/note="For pBluescript vector"
primer_bind complement(6993..7010)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 6999..7223
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
misc_feature 7311..11651
/label=Rosa26 right arm
/note="right homology arm from intron 1 of the mouse Rosa26
gene (Soriano, 1999)"
primer_bind complement(11736..11759)
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
primer_bind complement(11862..11882)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(11862..11880)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(11901..11917)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(11901..11917)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(11914..11936)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 11925..11941
/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(11949..11979)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 11994..12015
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(12132..12149)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(12303..12891)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(12383..12402)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(13062..13922)
/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 13685..13704
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(13923..14027)
/gene="bla"
/label=AmpR promoter
rep_origin complement(14053..14508)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(14190..14211)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 14402..14421
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 14635..14657
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 14649..14666
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 14650..14666
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 14676..14695
/label=T7
/note="T7 promoter, forward primer"
promoter 14676..14694
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 ccccgcggca ggccctccga gcgtggtgga gccgttctgt gagacagccg ggtacgagtc
61 gtgacgctgg aaggggcaag cgggtggtgg gcaggaatgc ggtccgccct gcagcaaccg
121 gagggggagg gagaagggag cggaaaagtc tccaccggac gcggccatgg ctcggggggg
181 ggggggcagc ggaggagcgc ttccggccga cgtctcgtcg ctgattggct tcttttcctc
241 ccgccgtgtg tgaaaacaca aatggcgtgt tttggttggc gtaaggcgcc tgtcagttaa
301 cggcagccgg agtgcgcagc cgccggcagc ctcgctctgc ccactgggtg gggcgggagg
361 taggtggggt gaggcgagct ggacgtgcgg gcgcggtcgg cctctggcgg ggcgggggag
421 gggagggagg gtcagcgaaa gtagctcgcg cgcgagcggc cgcccaccct ccccttcctc
481 tgggggagtc gttttacccg ccgccggccg ggcctcgtcg tctgattggc tctcggggcc
541 cagaaaactg gcccttgcca ttggctcgtg ttcgtgcaag ttgagtccat ccgccggcca
601 gcgggggcgg cgaggaggcg ctcccaggtt ccggccctcc cctcggcccc gcgccgcaga
661 gtctggccgc gcgcccctgc gcaacgtggc aggaagcgcg cgctgggggc ggggacgggc
721 agtagggctg agcggctgcg gggcgggtgc aagcacgttt ccgacttgag ttgcctcaag
781 aggggcgtgc tgagccagac ctccatcgcg cactccgggg agtggaggga aggagcgagg
841 gctcagttgg gctgttttgg aggcaggaag cacttgctct cccaaagtcg ctctgagttg
901 ttatcagtaa gggagctgca gtggagtagg cggggagaag gccgcaccct tctccggagg
961 ggggagggga gtgttgcaat acctttctgg gagttctctg ctgcctcctg gcttctgagg
1021 accgccctgg gcctgggaga atcccttccc cctcttccct cgtgatctgc aactccagtc
1081 tttctagcat ctgtagggcg cagtagtcca gggtttcctt gatgatgtca tacttatcct
1141 gtcccttttt tttccacagc tcgcggttga ggacaaactc ttcgcggtct ttccagtggt
1201 taattaaata acttcgtata gcatacatta tacgaagtta tggatccgaa caaacgaccc
1261 aacacccgtg cgttttattc tgtcttttta ttgccgatcc cctcagaaga actcgtcaag
1321 aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa
1381 gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc
1441 ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt
1501 ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc
1561 gggcatgcgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc
1621 gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg
1681 atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat
1741 tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg
1801 ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac
1861 agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag
1921 ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga
1981 cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa
2041 tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatggccga
2101 tcccatattg gctgcagggt cgctcggtgt tcgaggccac acgcgtcacc ttaatatgcg
2161 aagtggacct gggaccgcgc cgccccgact gcatctgcgt gttcgaattc gccaatgaca
2221 agacgctggg cggggtttgc tcgacattgg gtggaaacat tccaggcctg ggtggagagg
2281 ctttttgctt cctcttgcaa aaccacactg ctcgacattg ggtggaaaca ttccaggcct
2341 gggtggagag gctttttgct tcctcttgca aaaccacact gctcgactag tgattaaagt
2401 cgactcgggg acaccaaata tggcgatctc ggccttttcg tttcttggag ctgggacatg
2461 tttgccatcg atccatctac caccagaacg gccgttagat ctgctgccac cgttgtttcc
2521 accgaagaaa ccaccgttgc cgtaaccacc acgacggttg ttgctaaaga agctgccacc
2581 gccacggcca ccgttgtagc cgccgttgtt gttattgtag ttgctcatgt tatttctggc
2641 acttcttggt tttcctctta agtgaggagg aacataacca ttctcgttgt tgtcgttgat
2701 gcttaaattt tgcacttgtt cgctcagttc agccataata tgaaatgctt ttcttgttgt
2761 tcttacggaa taccacttgc cacctatcac cacaactaac tttttcccgt tcctccatct
2821 cttttatatt ttttttctcg agggatcttt gtgaaggaac cttacttctg tggtgtgaca
2881 taattggaca aactacctac agagatttaa agctctaagg taaatataaa atttttaagt
2941 gtataatgtg ttaaactact gattctaatt gtttgtgtat tttagattcc aacctatgga
3001 actgatgaat gggagcagtg gtggaatgcc tttaatgagg aaaacctgtt ttgctcagaa
3061 gaaatgccat ctagtgatga tgaggctact gctgactctc aacattctac tcctccaaaa
3121 aagaagagaa aggtagaaga ccccaaggac tttccttcag aattgctaag ttttttgagt
3181 catgctgtgt ttagtaatag aactcttgct tgctttgcta tttacaccac aaaggaaaaa
3241 gctgcactgc tatacaagaa aattatggaa aaatattctg taacctttat aagtaggcat
3301 aacagttata atcataacat actgtttttt cttactccac acaggcatag agtgtctgct
3361 attaataact atgctcaaaa attgtgtacc tttagctttt taatttgtaa aggggttaat
3421 aaggaatatt tgatgtatag tgccttgact agagatcata atcagccata ccacatttgt
3481 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
3541 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
3601 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
3661 caaactcatc aatgtatctt atcatgtctg gatctgacat ggtaagtaag cttataactt
3721 cgtatagcat acattatacg aagttatcgg cgcgccatca caagtttgta caaaaaagct
3781 gaacgagaaa cgtaaaatga tataaatatc aatatattaa attagatttt gcataaaaaa
3841 cagactacat aatactgtaa aacacaacat atccagtcat attggcggcc gcattaggca
3901 ccccaggctt tacactttat gcttccggct cgtataatgt gtggattttg agttaggatc
3961 cgtcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca
4021 ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc
4081 aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga
4141 aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc
4201 atccggaatt ccgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc
4261 cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc
4321 acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa
4381 acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtctca gccaatccct
4441 gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg
4501 ttttcaccat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc
4561 aggttcatca tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac
4621 agtactgcga tgagtggcag ggcggggcgt aaacgcgtgg atccggctta ctaaaagcca
4681 gataacagta tgcgtatttg cgcgctgatt tttgcggtat aagaatatat actgatatgt
4741 atacccgaag tatgtcaaaa agaggtatgc tatgaagcag cgtattacag tgacagttga
4801 cagcgacagc tatcagttgc tcaaggcata tatgatgtca atatctccgg tctggtaagc
4861 acaaccatgc agaatgaagc ccgtcgtctg cgtgccgaac gctggaaagc ggaaaatcag
4921 gaagggatgg ctgaggtcgc ccggtttatt gaaatgaacg gctcttttgc tgacgagaac
4981 aggggctggt gaaatgcagt ttaaggttta cacctataaa agagagagcc gttatcgtct
5041 gtttgtggat gtacagagtg atattattga cacgcccggg cgacggatgg tgatccccct
5101 ggccagtgca cgtctgctgt cagataaagt ctcccgtgaa ctttacccgg tggtgcatat
5161 cggggatgaa agctggcgca tgatgaccac cgatatggcc agtgtgccgg tctccgttat
5221 cggggaagaa gtggctgatc tcagccaccg cgaaaatgac atcaaaaacg ccattaacct
5281 gatgttctgg ggaatataaa tgtcaggctc ccttatacac agccagtctg caggtcgacc
5341 atagtgactg gatatgttgt gttttacagc attatgtagt ctgtttttta tgcaaaatct
5401 aatttaatat attgatattt atatcatttt acgtttctcg ttcagctttc ttgtacaaag
5461 tggtgatggc gcgccgcggc cgcgatcaat tcggtaccga agttcctatt ccgaagttcc
5521 tattctctag aaagtatagg aacttcctcg agggtacccc aattccgccc cccccccccc
5581 cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta
5641 ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc
5701 ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat
5761 gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc
5821 ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt
5881 gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt
5941 gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag
6001 aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt
6061 tagtcgaggt taaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa
6121 aaacacgatg ataatatggc cacaaccatg gtgagcaagg gcgaggagct gttcaccggg
6181 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
6241 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
6301 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
6361 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
6421 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
6481 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
6541 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
6601 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
6661 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
6721 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
6781 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
6841 ctcggcatgg acgagctgta caagtaatcc gggatccgga gagctcccaa cgaagttcct
6901 attccgaagt tcctattctc tagaaagtat aggaacttcc tcgaggttgg atgcagcccg
6961 ggggatccac tagttctaga gcggccgatc agcctcgact gtgccttcta gttgccagcc
7021 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
7081 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
7141 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
7201 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctcgagatc
7261 cactagttct agcctcgagg ctagagcggc cgccgcggat atcgaattcg ctagaagatg
7321 ggcgggagtc ttctgggcag gcttaaaggc taacctggtg tgtgggcgtt gtcctgcagg
7381 ggaattgaac aggtgtaaaa ttggagggac aagacttccc acagattttc ggttttgtcg
7441 ggaagttttt taataggggc aaataaggaa aatgggagga taggtagtca tctggggttt
7501 tatgcagcaa aactacaggt tattattgct tgtgatccgc ctcggagtat tttccatcga
7561 ggtagattaa agacatgctc acccgagttt tatactctcc tgcttgagat ccttactaca
7621 gtatgaaatt acagtgtcgc gagttagact atgtaagcag aattttaatc atttttaaag
7681 agcccagtac ttcatatcca tttctcccgc tccttctgca gccttatcaa aaggtatttt
7741 agaacactca ttttagcccc attttcattt attatactgg cttatccaac ccctagacag
7801 agcattggca ttttcccttt cctgatctta gaagtctgat gactcatgaa accagacaga
7861 ttagttacat acaccacaaa tcgaggctgt agctggggcc tcaacactgc agttctttta
7921 taactcctta gtacactttt tgttgatcct ttgccttgat ccttaatttt cagtgtctat
7981 cacctctccc gtcaggtggt gttccacatt tgggcctatt ctcagtccag ggagttttac
8041 aacaatagat gtattgagaa tccaacctaa agcttaactt tccactccca tgaatgcctc
8101 tctccttttt ctccatttat aaactgagct attaaccatt aatggtttcc aggtggatgt
8161 ctcctccccc aatattacct gatgtatctt acatattgcc aggctgatat tttaagacat
8221 taaaaggtat atttcattat tgagccacat ggtattgatt actgcttact aaaattttgt
8281 cattgtacac atctgtaaaa ggtggttcct tttggaatgc aaagttcagg tgtttgttgt
8341 ctttcctgac ctaaggtctt gtgagcttgt attttttcta tttaagcagt gctttctctt
8401 ggactggctt gactcatggc attctacacg ttattgctgg tctaaatgtg attttgccaa
8461 gcttcttcag gacctataat tttgcttgac ttgtagccaa acacaagtaa aatgattaag
8521 caacaaatgt atttgtgaag cttggttttt aggttgttgt gttgtgtgtg cttgtgctct
8581 ataataatac tatccagggg ctggagaggt ggctcggagt tcaagagcac agactgctct
8641 tccagaagtc ctgagttcaa ttcccagcaa ccacatggtg gctcacaacc atctgtaatg
8701 ggatctgatg ccctcttctg gtgtgtctga agaccacaag tgtattcaca ttaaataaat
8761 aaatcctcct tcttcttctt tttttttttt ttaaagagaa tactgtctcc agtagaattt
8821 actgaagtaa tgaaatactt tgtgtttgtt ccaatatggt agccaataat caaattactc
8881 tttaagcact ggaaatgtta ccaaggaact aatttttatt tgaagtgtaa ctgtggacag
8941 aggagccata actgcagact tgtgggatac agaagaccaa tgcagacttt aatgtctttt
9001 ctcttacact aagcaataaa gaaataaaaa ttgaacttct agtatcctat ttgtttaaac
9061 tgctagcttt acttaacttt tgtgcttcat ctatacaaag ctgaaagcta agtctgcagc
9121 cattactaaa catgaaagca agtaatgata attttggatt tcaaaaatgt agggccagag
9181 tttagccagc cagtggtggt gcttgccttt atgcctttaa tcccagcact ctggaggcag
9241 agacaggcag atctctgagt ttgagcccag cctggtctac acatcaagtt ctatctagga
9301 tagccaggaa tacacacaga aaccctgttg gggagggggg ctctgagatt tcataaaatt
9361 ataattgaag cattccctaa tgagccacta tggatgtggc taaatccgtc tacctttctg
9421 atgagatttg ggtattattt tttctgtctc tgctgttggt tgggtctttt gacactgtgg
9481 gctttcttta aagcctcctt cctgccatgt ggtctcttgt ttgctactaa cttcccatgg
9541 cttaaatggc atggcttttt gccttctaag ggcagctgct gagatttgca gcctgatttc
9601 cagggtgggg ttgggaaatc tttcaaacac taaaattgtc ctttaatttt ttttttaaaa
9661 aatgggttat ataataaacc tcataaaata gttatgagga gtgaggtgga ctaatattaa
9721 atgagtccct cccctataaa agagctatta aggctttttg tcttatactt aacttttttt
9781 ttaaatgtgg tatctttaga accaagggtc ttagagtttt agtatacaga aactgttgca
9841 tcgcttaatc agattttcta gtttcaaatc cagagaatcc aaattcttca cagccaaagt
9901 caaattaaga atttctgact tttaatgtta atttgcttac tgtgaatata aaaatgatag
9961 cttttcctga ggcagggtct cactatgtat ctctgcctga tctgcaacaa gatatgtaga
10021 ctaaagttct gcctgctttt gtctcctgaa tactaaggtt aaaatgtagt aatacttttg
10081 gaacttgcag gtcagattct tttatagggg acacactaag ggagcttggg tgatagttgg
10141 taaaatgtgt ttcaagtgat gaaaacttga attattatca ccgcaaccta ctttttaaaa
10201 aaaaaagcca ggcctgttag agcatgctta agggatccct aggacttgct gagcacacaa
10261 gagtagttac ttggcaggct cctggtgaga gcatatttca aaaaacaagg cagacaacca
10321 agaaactaca gttaaggtta cctgtcttta aaccatctgc atatacacag ggatattaaa
10381 atattccaaa taatatttca ttcaagtttt cccccatcaa attgggacat ggatttctcc
10441 ggtgaatagg cagagttgga aactaaacaa atgttggttt tgtgatttgt gaaattgttt
10501 tcaagtgata gttaaagccc atgagataca gaacaaagct gctatttcga ggtctcttgg
10561 tttatactca gaagcacttc tttgggtttc cctgcactat cctgatcatg tgctaggcct
10621 accttaggct gattgttgtt caaataaact taagtttcct gtcaggtgat gtcatatgat
10681 ttcatatatc aaggcaaaac atgttatata tgttaaacat ttgtacttaa tgtgaaagtt
10741 aggtctttgt gggtttgatt tttaattttc aaaacctgag ctaaataagt catttttaca
10801 tgtcttacat ttggtggaat tgtataattg tggtttgcag gcaagactct ctgacctagt
10861 aaccctacct atagagcact ttgctgggtc acaagtctag gagtcaagca tttcaccttg
10921 aagttgagac gttttgttag tgtatactag tttatatgtt ggaggacatg tttatccaga
10981 agatattcag gactattttt gactgggcta aggaattgat tctgattagc actgttagtg
11041 agcattgagt ggcctttagg cttgaattgg agtcacttgt atatctcaaa taatgctggc
11101 cttttttaaa aagcccttgt tctttatcac cctgttttct acataatttt tgttcaaaga
11161 aatacttgtt tggatctcct tttgacaaca atagcatgtt ttcaagccat attttttttc
11221 cttttttttt ttttttttgg tttttcgaga cagggtttct ctgtatagcc ctggctgtcc
11281 tggaactcac tttgtagacc aggctggcct cgaactcaga aatccgcctg cctctgcctc
11341 ctgagtgccg ggattaaagg cgtgcaccac cacgcctggc taagttggat attttgttat
11401 ataactataa ccaatactaa ctccactggg tggattttta attcagtcag tagtcttaag
11461 tggtctttat tggcccttca ttaaaatcta ctgttcactc taacagaggc tgttggtact
11521 agtggcactt aagcaacttc ctacggatat actagcagat taagggtcag ggatagaaac
11581 tagtctagcg ttttgtatac ctaccagctt tatactacct tgttctgata gaaatatttc
11641 aggacatcta gcttatcgat accgtcgacg gtatcgataa gcttgatatc gaattctacc
11701 gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg
11761 gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc
11821 gccgccgccc tgcaggcgat cgctacgtac agcttttgtt ccctttagtg agggttaatt
11881 gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
11941 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
12001 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
12061 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
12121 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
12181 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
12241 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
12301 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
12361 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
12421 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
12481 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
12541 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
12601 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
12661 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
12721 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
12781 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
12841 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
12901 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
12961 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
13021 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
13081 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
13141 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
13201 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
13261 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
13321 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
13381 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
13441 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
13501 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
13561 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
13621 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
13681 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
13741 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
13801 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
13861 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
13921 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
13981 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
14041 aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
14101 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
14161 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
14221 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
14281 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
14341 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
14401 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
14461 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat
14521 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
14581 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
14641 cacgacgttg taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat
14701 tggagct
//
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