| 质粒名称: | pR26 CAG/GFP Dest |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 74281 |
| 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: | 16896 bp |
| 载体类型: | Mouse Targeting |
| 载体抗性: | Chloramphenicol and Ampicillin |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 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 CAG/GFP Dest was a gift from Ralf Kuehn (Addgene plasmid # 74281 ; http://n2t.net/addgene:74281 ; RRID:Addgene_74281) |
| 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
pR26 CAG/GFP Dest载体质粒基本信息
pR26 CAG/GFP Dest质粒图谱载体图谱和pR26 CAG/GFP Dest载体序列质粒序列多克隆位点信息
pR26 CAG/GFP Dest质粒载体简介
pR26 CAG/GFP Dest质粒序列载体序列
LOCUS pR26 CAG/GFP Dest 16896 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Gene targeting vector for the mouse Rosa26 locus, including a CAG
promoter, loxP flanked stop cassette, GFP, for gateway cloning.
ACCESSION .
VERSION .
KEYWORDS pR26 CAG/GFP Dest
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 16896)
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 16896)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..16896
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 6..1088
/label=Rosa26 left arm
/note="left homology arm from intron 1 of the mouse Rosa26
gene (Soriano, 1999)"
misc_feature 1095..1198
/label=Ad SA
/note="adenovirus major late transcript splice acceptor
(Friedrich and Soriano, 1991)"
polyA_signal 1217..1424
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
polyA_signal 1438..1645
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
enhancer 1654..2033
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
intron 2309..3325
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 3249..3271
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 3333..3352
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
protein_bind 3388..3421
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
polyA_signal 3428..3476
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
CDS complement(3483..4286)
/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(3594..3613)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 4204..4223
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
intron 5099..5164
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
polyA_signal 5739..5873
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5776..5795)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5830..5849
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
protein_bind 5893..5926
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
protein_bind 5939..6063
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 6088..6118
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 6172..6831
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(6238..6257)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
CDS 7173..7478
/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 7357..7376
/label=ccdB-fwd
/note="ccdB gene, forward primer"
protein_bind complement(7519..7643)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
protein_bind 7678..7725
/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 7764..8316
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(7908..7925)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 8135..8154
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 8328..9047
/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 8328..9044
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(8373..8394)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(8634..8653)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 8981..9002
/label=EGFP-C
/note="EGFP, forward primer"
protein_bind 9072..9119
/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(9144..9160)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(9144..9160)
/label=pBluescriptSK
/note="For pBluescript vector"
primer_bind complement(9173..9190)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 9179..9403
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
misc_feature 9493..13833
/label=Rosa26 right arm
/note="right homology arm from intron 1 of the mouse Rosa26
gene (Soriano, 1999)"
primer_bind complement(13918..13941)
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
primer_bind complement(14051..14071)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(14051..14069)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(14090..14106)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(14090..14106)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(14103..14125)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 14114..14130
/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(14138..14168)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 14183..14204
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(14321..14338)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(14492..15080)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(14572..14591)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(15251..16111)
/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 15874..15893
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(16112..16216)
/gene="bla"
/label=AmpR promoter
rep_origin complement(16269..16697)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(16379..16400)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 16591..16610
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 16824..16846
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 16838..16855
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 16839..16855
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 16865..16884
/label=T7
/note="T7 promoter, forward primer"
promoter 16865..16883
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 ccaccgcggc aggccctccg agcgtggtgg agccgttctg tgagacagcc gggtacgagt
61 cgtgacgctg gaaggggcaa gcgggtggtg ggcaggaatg cggtccgccc tgcagcaacc
121 ggagggggag ggagaaggga gcggaaaagt ctccaccgga cgcggccatg gctcgggggg
181 gggggggcag cggaggagcg cttccggccg acgtctcgtc gctgattggc ttcttttcct
241 cccgccgtgt gtgaaaacac aaatggcgtg ttttggttgg cgtaaggcgc ctgtcagtta
301 acggcagccg gagtgcgcag ccgccggcag cctcgctctg cccactgggt ggggcgggag
361 gtaggtgggg tgaggcgagc tggacgtgcg ggcgcggtcg gcctctggcg gggcgggggg
421 ggggagggag ggtcagcgaa agtagctcgc gcgcgagcgg ccgcccaccc tccccttcct
481 ctgggggagt cgttttaccc gccgccggcc gggcctcgtc gtctgattgg ctctcggggc
541 ccagaaaact ggcccttgcc attggctcgt gttcgtgcaa gttgagtcca tccgccggcc
601 agcgggggcg gcgaggaggc gctcccaggt tccggccctc ccctcggccc cgcgccgcag
661 agtctggccg cgcgcccctg cgcaacgtgg caggaagcgc gcgctggggg cggggacggg
721 cagtagggct gagcggctgc ggggcgggtg caagcacgtt tccgacttga gttgcctcaa
781 gaggggcgtg ctgagccaga cctccatcgc gcactccggg gagtggaggg aaggagcgag
841 ggctcagttg ggctgttttg gaggcaggaa gcacttgctc tcccaaagtc gctctgagtt
901 gttatcagta agggagctgc agtggagtag gcggggagaa ggccgcaccc ttctccggag
961 gggggagggg agtgttgcaa tacctttctg ggagttctct gctgcctcct ggcttctgag
1021 gaccgccctg ggcctgggag aatcccttcc ccctcttccc tcgtgatctg caactccagt
1081 ctttctagca tctgtagggc gcagtagtcc agggtttcct tgatgatgtc atacttatcc
1141 tgtccctttt ttttccacag ctcgcggttg aggacaaact cttcgcggtc tttccagtgg
1201 ttaattaact cgaggactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
1261 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
1321 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
1381 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt ctagcgactg
1441 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
1501 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
1561 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
1621 aagacaatag caggcatgct ggggatgcgg gtcgacattg attattgact agttattaat
1681 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
1741 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
1801 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
1861 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
1921 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
1981 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg
2041 tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt
2101 atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg ggcgcgcgcc
2161 aggcgggggg gggggggggg gggggggggg gggggggggg gggggggggg cggcggcagc
2221 caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc
2281 ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc
2341 ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca
2401 ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg
2461 gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg
2521 cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg
2581 ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg
2641 cagtgtgcgc gaggggagcg cggccggggg cggtgccccg cggtgcgggg ggggctgcga
2701 ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc
2761 gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga gttgctgagc acggcccggc
2821 ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg
2881 gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggagg gctcggggga
2941 ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg
3001 ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctgtgcgga
3061 gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga agcggtgcgg
3121 cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct
3181 tctccctctc cagcctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg
3241 cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg ctaaccatgt
3301 tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg tgctgtctca
3361 tcattttggc aaagaattct taattaaata acttcgtata gcatacatta tacgaagtta
3421 tggatccgaa caaacgaccc aacacccgtg cgttttattc tgtcttttta ttgccgatcc
3481 cctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg
3541 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca
3601 cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg
3661 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc
3721 acgacgagat cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggc
3781 gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga
3841 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca
3901 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg
3961 tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct
4021 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc
4081 cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga
4141 accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt
4201 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat
4261 ccatcttgtt caatggccga tcccatattg gctgcagggt cgctcggtgt tcgaggccac
4321 acgcgtcacc ttaatatgcg aagtggacct gggaccgcgc cgccccgact gcatctgcgt
4381 gttcgaattc gccaatgaca agacgctggg cggggtttgc tcgacattgg gtggaaacat
4441 tccaggcctg ggtggagagg ctttttgctt cctcttgcaa aaccacactg ctcgacattg
4501 ggtggaaaca ttccaggcct gggtggagag gctttttgct tcctcttgaa aaccacactg
4561 ctcgactagt gattaaagtc gactcgggga caccaaatat ggcgatctcg gccttttcgt
4621 ttcttggagc tgggacatgt ttgccatcga tccatctacc accagaacgg ccgttagatc
4681 tgctgccacc gttgtttcca ccgaagaaac caccgttgcc gtaaccacca cgacggttgt
4741 tgctaaagaa gctgccaccg ccacggccac cgttgtagcc gccgttgttg ttattgtagt
4801 tgctactgtt atttctggca cttcttggtt ttcctcttaa gtgaggagga acataaccat
4861 tctcgttgtt gtcgttgatg cttaaatttt gcacttgttc gctcagttca gccataatat
4921 gaaatgcttt tcttgttgtt cttacggaat accacttgcc acctatcacc acaactaact
4981 ttttcccgtt cctccatctc ttttatattt tttttctcga gggatctttg tgaaggaacc
5041 ttacttctgt ggtgtgacat aattggacaa actacctaca gagatttaaa gctctaaggt
5101 aaatataaaa tttttaagtg tataatgtgt taaactactg attctaattg tttgtgtatt
5161 ttagattcca acctatggaa ctgatgaatg ggagcagtgg tggaatgcct ttaatgagga
5221 aaacctgttt tgctcagaag aaatgccatc tagtgatgat gaggctactg ctgactctca
5281 acattctact cctccaaaaa agaagagaag ggtagaagac cccaaggact ttccttcaga
5341 attgctaagt tttttgagtc atgctgtgtt tagtaataga actcttgctt gctttgctat
5401 ttacaccaca aaggaaaaag ctgcactgct atacaagaaa attatggaaa aatattctgt
5461 aacctttata agtaggcata acagttataa tcataacata ctgttttttc ttactccaca
5521 caggcataga gtgtctgcta ttaataacta tgctcaaaaa ttgtgtacct ttagcttttt
5581 aatttgtaaa ggggttaata aggaatattt gatgtatagt gccttgacta gagatcataa
5641 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
5701 tgaacctgaa acatagaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
5761 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
5821 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atctgacatg
5881 gtaagtaagc ttataacttc gtatagcata cattatacga agttatcggc gcgccatcac
5941 aagtttgtac aaaaaagctg aacgagaaac gtaaaatgat ataaatatca atatattaaa
6001 ttagattttg cataaaaaac agactacata atactgtaaa acacaacata tccagtcact
6061 atggcggccg cattaggcac cccaggcttt acactttatg cttccggctc gtataatgtg
6121 tggattttga gttaggatcc gtcgagattt tcaggagcta aggaagctaa aatggagaaa
6181 aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag
6241 gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc
6301 tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt
6361 gcccgcctga tgaatgctca tccggaattc cgtatggcaa tgaaagacgg tgagctggtg
6421 atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca
6481 tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat
6541 gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt
6601 ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg
6661 gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg
6721 ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga
6781 atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta aacgcgtgga
6841 tccggcttac taaaagccag ataacagtat gcgtatttgc gcgctgattt ttgcggtata
6901 agaatatata ctgatatgta tacccgaagt atgtcaaaaa gaggtatgct atgaagcagc
6961 gtattacagt gacagttgac agcgacagct atcagttgct caaggcatat atgatgtcaa
7021 tatctccggt ctggtaagca caaccatgca gaatgaagcc cgtcgtctgc gtgccgaacg
7081 ctggaaagcg gaaaatcagg aagggatggc tgaggtcgcc cggtttattg aaatgaacgg
7141 ctcttttgct gacgagaaca ggggctggtg aaatgcagtt taaggtttac acctataaaa
7201 gagagagccg ttatcgtctg tttgtggatg tacagagtga tattattgac acgcccgggc
7261 gacggatggt gatccccctg gccagtgcac gtctgctgtc agataaagtc tcccgtgaac
7321 tttacccggt ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc gatatggcca
7381 gtgtgccggt ctccgttatc ggggaagaag tggctgatct cagccaccgc gaaaatgaca
7441 tcaaaaacgc cattaacctg atgttctggg gaatataaat gtcaggctcc cttatacaca
7501 gccagtctgc aggtcgacca tagtgactgg atatgttgtg ttttacagta ttatgtagtc
7561 tgttttttat gcaaaatcta atttaatata ttgatattta tatcatttta cgtttctcgt
7621 tcagctttct tgtacaaagt ggtgatggcg cgccgcggcc gcgatcaatt cggtaccgaa
7681 gttcctattc cgaagttcct attctctaga aagtatagga acttcctcga gggtacccca
7741 attccgcccc cccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg
7801 tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg
7861 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg
7921 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca
7981 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct
8041 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca
8101 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa
8161 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg
8221 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg
8281 ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg gtgagcaagg
8341 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
8401 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
8461 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
8521 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
8581 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
8641 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
8701 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
8761 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
8821 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
8881 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
8941 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
9001 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaatcc gggatccgga
9061 gagctcccaa cgaagttcct attccgaagt tcctattctc tagaaagtat aggaacttcc
9121 tcgaggttgg atgcagcccg ggggatccac tagttctaga gcggccgatc agcctcgact
9181 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
9241 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
9301 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
9361 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
9421 accagctggg gctcgagatc cactagttct agcctcgagg ctagagcggc cgccgcgccg
9481 atatcgaatt cgctagaaga tgggcgggag tcttctgggc aggcttaaag gctaacctgg
9541 tgtgtgggcg ttgtcctgca ggggaattga acaggtgtaa aattggaggg acaagacttc
9601 ccacagattt tcggttttgt cgggaagttt tttaataggg gcaaataagg aaaatgggag
9661 gataggtagt catctggggt tttatgcagc aaaactacag gttattattg cttgtgatcc
9721 gcctcggagt attttccatc gaggtagatt aaagacatgc tcacccgagt tttatactct
9781 cctgcttgag atccttacta cagtatgaaa ttacagtgtc gcgagttaga ctatgtaagc
9841 agaattttaa tcatttttaa agagcccagt acttcatatc catttctccc gctccttctg
9901 cagccttatc aaaaggtatt ttagaacact cattttagcc ccattttcat ttattatact
9961 ggcttatcca acccctagac agagcattgg cattttccct ttcctgatct tagaagtctg
10021 atgactcatg aaaccagaca gattagttac atacaccaca aatcgaggct gtagctgggg
10081 cctcaacact gcagttcttt tataactcct tagtacactt tttgttgatc ctttgccttg
10141 atccttaatt ttcagtgtct atcacctctc ccgtcaggtg gtgttccaca tttgggccta
10201 ttctcagtcc agggagtttt acaacaatag atgtattgag aatccaacct aaagcttaac
10261 tttccactcc catgaatgcc tctctccttt ttctccattt ataaactgag ctattaacca
10321 ttaatggttt ccaggtggat gtctcctccc ccaatattac ctgatgtatc ttacatattg
10381 ccaggctgat attttaagac attaaaaggt atatttcatt attgagccac atggtattga
10441 ttactgctta ctaaaatttt gtcattgtac acatctgtaa aaggtggttc cttttggaat
10501 gcaaagttca ggtgtttgtt gtctttcctg acctaaggtc ttgtgagctt gtattttttc
10561 tatttaagca gtgctttctc ttggactggc ttgactcatg gcattctaca cgttattgct
10621 ggtctaaatg tgattttgcc aagcttcttc aggacctata attttgcttg acttgtagcc
10681 aaacacaagt aaaatgatta agcaacaaat gtatttgtga agcttggttt ttaggttgtt
10741 gtgttgtgtg tgcttgtgct ctataataat actatccagg ggctggagag gtggctcgga
10801 gttcaagagc acagactgct cttccagaag tcctgagttc aattcccagc aaccacatgg
10861 tggctcacaa ccatctgtaa tgggatctga tgccctcttc tggtgtgtct gaagaccaca
10921 agtgtattca cattaaataa ataaatcctc cttcttcttc tttttttttt ttttaaagag
10981 aatactgtct ccagtagaat ttactgaagt aatgaaatac tttgtgtttg ttccaatatg
11041 gtagccaata atcaaattac tctttaagca ctggaaatgt taccaaggaa ctaattttta
11101 tttgaagtgt aactgtggac agaggagcca taactgcaga cttgtgggat acagaagacc
11161 aatgcagact ttaatgtctt ttctcttaca ctaagcaata aagaaataaa aattgaactt
11221 ctagtatcct atttgtttaa actgctagct ttacttaact tttgtgcttc atctatacaa
11281 agctgaaagc taagtctgca gccattacta aacatgaaag caagtaatga taattttgga
11341 tttcaaaaat gtagggccag agtttagcca gccagtggtg gtgcttgcct ttatgccttt
11401 aatcccagca ctctggaggc agagacaggc agatctctga gtttgagccc agcctggtct
11461 acacatcaag ttctatctag gatagccagg aatacacaca gaaaccctgt tggggagggg
11521 ggctctgaga tttcataaaa ttataattga agcattccct aatgagccac tatggatgtg
11581 gctaaatccg tctacctttc tgatgagatt tgggtattat tttttctgtc tctgctgttg
11641 gttgggtctt ttgacactgt gggctttctt taaagcctcc ttcctgccat gtggtctctt
11701 gtttgctact aacttcccat ggcttaaatg gcatggcttt ttgccttcta agggcagctg
11761 ctgagatttg cagcctgatt tccagggtgg ggttgggaaa tctttcaaac actaaaattg
11821 tcctttaatt ttttttttaa aaaatgggtt atataataaa cctcataaaa tagttatgag
11881 gagtgaggtg gactaatatt aaatgagtcc ctcccctata aaagagctat taaggctttt
11941 tgtcttatac ttaacttttt ttttaaatgt ggtatcttta gaaccaaggg tcttagagtt
12001 ttagtataca gaaactgttg catcgcttaa tcagattttc tagtttcaaa tccagagaat
12061 ccaaattctt cacagccaaa gtcaaattaa gaatttctga cttttaatgt taatttgctt
12121 actgtgaata taaaaatgat agcttttcct gaggcagggt ctcactatgt atctctgcct
12181 gatctgcaac aagatatgta gactaaagtt ctgcctgctt ttgtctcctg aatactaagg
12241 ttaaaatgta gtaatacttt tggaacttgc aggtcagatt cttttatagg ggacacacta
12301 agggagcttg ggtgatagtt ggtaaaatgt gtttcaagtg atgaaaactt gaattattat
12361 caccgcaacc tactttttaa aaaaaaaagc caggcctgtt agagcatgct taagggatcc
12421 ctaggacttg ctgagcacac aagagtagtt acttggcagg ctcctggtga gagcatattt
12481 caaaaaacaa ggcagacaac caagaaacta cagttaaggt tacctgtctt taaaccatct
12541 gcatatacac agggatatta aaatattcca aataatattt cattcaagtt ttcccccatc
12601 aaattgggac atggatttct ccggtgaata ggcagagttg gaaactaaac aaatgttggt
12661 tttgtgattt gtgaaattgt tttcaagtga tagttaaagc ccatgagata cagaacaaag
12721 ctgctatttc gaggtctctt ggtttatact cagaagcact tctttgggtt tccctgcact
12781 atcctgatca tgtgctaggc ctaccttagg ctgattgttg ttcaaataaa cttaagtttc
12841 ctgtcaggtg atgtcatatg atttcatata tcaaggcaaa acatgttata tatgttaaac
12901 atttgtactt aatgtgaaag ttaggtcttt gtgggtttga tttttaattt tcaaaacctg
12961 agctaaataa gtcattttta catgtcttac atttggtgga attgtataat tgtggtttgc
13021 aggcaagact ctctgaccta gtaaccctac ctatagagca ctttgctggg tcacaagtct
13081 aggagtcaag catttcacct tgaagttgag acgttttgtt agtgtatact agtttatatg
13141 ttggaggaca tgtttatcca gaagatattc aggactattt ttgactgggc taaggaattg
13201 attctgatta gcactgttag tgagcattga gtggccttta ggcttgaatt ggagtcactt
13261 gtatatctca aataatgctg gcctttttta aaaagccctt gttctttatc accctgtttt
13321 ctacataatt tttgttcaaa gaaatacttg tttggatctc cttttgacaa caatagcatg
13381 ttttcaagcc atattttttt tccttttttt tttttttttt ggtttttcga gacagggttt
13441 ctctgtatag ccctggctgt cctggaactc actttgtaga ccaggctggc ctcgaactca
13501 gaaatccgcc tgcctctgcc tcctgagtgc cgggattaaa ggcgtgcacc accacgcctg
13561 gctaagttgg atattttgtt atataactat aaccaatact aactccactg ggtggatttt
13621 taattcagtc agtagtctta agtggtcttt attggccctt cattaaaatc tactgttcac
13681 tctaacagag gctgttggta ctagtggcac ttaagcaact tcctacggat atactagcag
13741 attaagggtc agggatagaa actagtctag cgttttgtat acctaccagc tttatactac
13801 cttgttctga tagaaatatt tcaggacatc tagcttatcg ataccgtcga cggtatcgat
13861 aagcttgata tcgaattcta ccgggtaggg gaggcgcttt tcccaaggca gtctggagca
13921 tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg gcctcgcaca
13981 cattccacat ccaccggtag gcgccgccgc cctgcagggc gatcgcggcc gcggtaccca
14041 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
14101 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
14161 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
14221 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
14281 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
14341 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
14401 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
14461 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
14521 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
14581 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
14641 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
14701 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
14761 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
14821 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
14881 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
14941 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
15001 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
15061 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
15121 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
15181 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
15241 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
15301 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
15361 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
15421 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
15481 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
15541 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
15601 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
15661 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
15721 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
15781 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
15841 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
15901 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
15961 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
16021 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
16081 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
16141 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
16201 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
16261 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
16321 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
16381 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
16441 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
16501 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
16561 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
16621 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
16681 gccgctacag ggcacgttcc attcgccatt caggctgcgc aactgttggg aagggcgatc
16741 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
16801 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg
16861 cgcgtaatac gactcactat agggcgaatt ggagct
//
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