| 质粒名称: | pR26 iBFP Dest |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 74284 |
| 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: | 14523 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 iBFP Dest was a gift from Ralf Kuehn (Addgene plasmid # 74284 ; http://n2t.net/addgene:74284 ; RRID:Addgene_74284) |
| 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 |
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 iBFP Dest 14523 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Gene targeting vector for the mouse Rosa26 locus, including a loxP
flanked stop cassette and BFP, for gateway cloning.
ACCESSION .
VERSION .
KEYWORDS pR26 iBFP Dest
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14523)
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 14523)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..14523
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 3..1085
/label=Rosa26 left arm
/note="left homology arm from intron 1 of the mouse Rosa26
gene (Soriano, 1999)"
misc_feature 1092..1195
/label=Ad SA
/note="adenovirus major late transcript splice acceptor
(Friedrich and Soriano, 1991)"
protein_bind 1206..1239
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
polyA_signal 1246..1294
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
CDS complement(1301..2104)
/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(1412..1431)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 2022..2041
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
intron 2918..2983
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 3113..3133
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 3558..3692
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3595..3614)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3649..3668
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
protein_bind 3712..3745
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
protein_bind 3758..3882
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 3907..3937
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 3991..4650
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(4057..4076)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
CDS 4992..5297
/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 5176..5195
/label=ccdB-fwd
/note="ccdB gene, forward primer"
protein_bind complement(5338..5462)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
protein_bind complement(5474..5507)
/label=FRT (minimal)
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
misc_feature 5544..6096
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(5688..5705)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 5915..5934
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 6117..6836
/codon_start=1
/product="enhanced blue variant of GFP (Ai et al., 2007)"
/label=EBFP2
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
EFRTAAGITLGMDELYK"
primer_bind complement(6162..6183)
/label=EGFP-N
/note="EGFP, reverse primer"
protein_bind complement(6844..6877)
/label=FRT (minimal)
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
polyA_signal 6886..7093
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
misc_feature 7132..11471
/label=Rosa26 right arm
/note="right homology arm from intron 1 of the mouse Rosa26
gene (Soriano, 1999)"
primer_bind complement(11676..11696)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(11676..11694)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(11715..11731)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(11715..11731)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(11728..11750)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 11739..11755
/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(11763..11793)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 11808..11829
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(11946..11963)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(12117..12705)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(12197..12216)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(12876..13736)
/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 13499..13518
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(13737..13841)
/gene="bla"
/label=AmpR promoter
rep_origin complement(13867..14322)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(14004..14025)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 14216..14235
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 14449..14471
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 14463..14480
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 14464..14480
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 14490..14509
/label=T7
/note="T7 promoter, forward primer"
promoter 14490..14508
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 ccgcggcagg ccctccgagc gtggtggagc cgttctgtga gacagccggg tacgagtcgt
61 gacgctggaa ggggcaagcg ggtggtgggc aggaatgcgg tccgccctgc agcaaccgga
121 gggggaggga gaagggagcg gaaaagtctc caccggacgc ggccatggct cggggggggg
181 ggggcagcgg aggagcgctt ccggccgacg tctcgtcgct gattggcttc ttttcctccc
241 gccgtgtgtg aaaacacaaa tggcgtgttt tggttggcgt aaggcgcctg tcagttaacg
301 gcagccggag tgcgcagccg ccggcagcct cgctctgccc actgggtggg gcgggaggta
361 ggtggggtga ggcgagctgg acgtgcgggc gcggtcggcc tctggcgggg cgggggaggg
421 gagggagggt cagcgaaagt agctcgcgcg cgagcggccg cccaccctcc ccttcctctg
481 ggggagtcgt tttacccgcc gccggccggg cctcgtcgtc tgattggctc tcggggccca
541 gaaaactggc ccttgccatt ggctcgtgtt cgtgcaagtt gagtccatcc gccggccagc
601 gggggcggcg aggaggcgct cccaggttcc ggccctcccc tcggccccgc gccgcagagt
661 ctggccgcgc gcccctgcgc aacgtggcag gaagcgcgcg ctgggggcgg ggacgggcag
721 tagggctgag cggctgcggg gcgggtgcaa gcacgtttcc gacttgagtt gcctcaagag
781 gggcgtgctg agccagacct ccatcgcgca ctccggggag tggagggaag gagcgagggc
841 tcagttgggc tgttttggag gcaggaagca cttgctctcc caaagtcgct ctgagttgtt
901 atcagtaagg gagctgcagt ggagtaggcg gggagaaggc cgcacccttc tccggagggg
961 ggaggggagt gttgcaatac ctttctggga gttctctgct gcctcctggc ttctgaggac
1021 cgccctgggc ctgggagaat cccttccccc tcttccctcg tgatctgcaa ctccagtctt
1081 tctagcatct gtagggcgca gtagtccagg gtttccttga tgatgtcata cttatcctgt
1141 cccttttttt tccacagctc gcggttgagg acaaactctt cgcggtcttt ccagtggtta
1201 attaaataac ttcgtatagc atacattata cgaagttatg gatccgaaca aacgacccaa
1261 cacccgtgcg ttttattctg tctttttatt gccgatcccc tcagaagaac tcgtcaagaa
1321 ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc
1381 ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct
1441 gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt
1501 ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg
1561 gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt
1621 ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat
1681 gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg
1741 catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc
1801 ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag
1861 ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcctgcagtt
1921 cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca
1981 gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata
2041 gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atggccgatc
2101 ccatattggc tgcagggtcg ctcggtgttc gaggccacac gcgtcacctt aatatgcgaa
2161 gtggacctgg gaccgcgccg ccccgactgc atctgcgtgt tcgaattcgc caatgacaag
2221 acgctgggcg gggtttgctc gacattgggt ggaaacattc caggcctggg tggagaggct
2281 ttttgcttcc tcttgcaaaa ccacactgct cgacattggg tggaaacatt ccaggcctgg
2341 gtggagaggc tttttgcttc ctcttgcaaa accacactgc tcgactagtg attaaagtcg
2401 actcggggac accaaatatg gcgatctcgg ccttttcgtt tcttggagct gggacatgtt
2461 tgccatcgat ccatctacca ccagaacggc cgttagatct gctgccaccg ttgtttccac
2521 cgaagaaacc accgttgccg taaccaccac gacggttgtt gctaaagaag ctgccaccgc
2581 cacggccacc gttgtagccg ccgttgttgt tattgtagtt gctcatgtta tttctggcac
2641 ttcttggttt tcctcttaag tgaggaggaa cataaccatt ctcgttgttg tcgttgatgc
2701 ttaaattttg cacttgttcg ctcagttcag ccataatatg aaatgctttt cttgttgttc
2761 ttacggaata ccacttgcca cctatcacca caactaactt tttcccgttc ctccatctct
2821 tttatatttt ttttctcgag ggatctttgt gaaggaacct tacttctgtg gtgtgacata
2881 attggacaaa ctacctacag agatttaaag ctctaaggta aatataaaat ttttaagtgt
2941 ataatgtgtt aaactactga ttctaattgt ttgtgtattt tagattccaa cctatggaac
3001 tgatgaatgg gagcagtggt ggaatgcctt taatgaggaa aacctgtttt gctcagaaga
3061 aatgccatct agtgatgatg aggctactgc tgactctcaa cattctactc ctccaaaaaa
3121 gaagagaaag gtagaagacc ccaaggactt tccttcagaa ttgctaagtt ttttgagtca
3181 tgctgtgttt agtaatagaa ctcttgcttg ctttgctatt tacaccacaa aggaaaaagc
3241 tgcactgcta tacaagaaaa ttatggaaaa atattctgta acctttataa gtaggcataa
3301 cagttataat cataacatac tgttttttct tactccacac aggcatagag tgtctgctat
3361 taataactat gctcaaaaat tgtgtacctt tagcttttta atttgtaaag gggttaataa
3421 ggaatatttg atgtatagtg ccttgactag agatcataat cagccatacc acatttgtag
3481 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa catagaatga
3541 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
3601 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
3661 aactcatcaa tgtatcttat catgtctgga tctgacatgg taagtaagct tataacttcg
3721 tatagcatac attatacgaa gttatcggcg cgccatcaca agtttgtaca aaaaagctga
3781 acgagaaacg taaaatgata taaatatcaa tatattaaat tagattttgc ataaaaaaca
3841 gactacataa tactgtaaaa cacaacatat ccagtcacta tggcggccgc attaggcacc
3901 ccaggcttta cactttatgc ttccggctcg tataatgtgt ggattttgag ttaggatccg
3961 tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc
4021 gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa
4081 tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa
4141 aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat
4201 ccggaattcc gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct
4261 tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac
4321 gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac
4381 ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg
4441 gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt
4501 ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag
4561 gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag
4621 tactgcgatg agtggcaggg cggggcgtaa acgcgtggat ccggcttact aaaagccaga
4681 taacagtatg cgtatttgcg cgctgatttt tgcggtataa gaatatatac tgatatgtat
4741 acccgaagta tgtcaaaaag aggtatgcta tgaagcagcg tattacagtg acagttgaca
4801 gcgacagcta tcagttgctc aaggcatata tgatgtcaat atctccggtc tggtaagcac
4861 aaccatgcag aatgaagccc gtcgtctgcg tgccgaacgc tggaaagcgg aaaatcagga
4921 agggatggct gaggtcgccc ggtttattga aatgaacggc tcttttgctg acgagaacag
4981 gggctggtga aatgcagttt aaggtttaca cctataaaag agagagccgt tatcgtctgt
5041 ttgtggatgt acagagtgat attattgaca cgcccgggcg acggatggtg atccccctgg
5101 ccagtgcacg tctgctgtca gataaagtct cccgtgaact ttacccggtg gtgcatatcg
5161 gggatgaaag ctggcgcatg atgaccaccg atatggccag tgtgccggtc tccgttatcg
5221 gggaagaagt ggctgatctc agccaccgcg aaaatgacat caaaaacgcc attaacctga
5281 tgttctgggg aatataaatg tcaggctccc ttatacacag ccagtctgca ggtcgaccat
5341 agtgactgga tatgttgtgt tttacagcat tatgtagtct gttttttatg caaaatctaa
5401 tttaatatat tgatatttat atcattttac gtttctcgtt cagctttctt gtacaaagtg
5461 gtgatggcgc gccgaagttc ctatactttc tagagaatag gaacttctct ctcgaggtta
5521 acgaattccg cccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg
5581 tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg
5641 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg
5701 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca
5761 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct
5821 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca
5881 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa
5941 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg
6001 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg
6061 ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaacccgg gacaccatgg
6121 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
6181 acgtaaacgg ccacaagttc agcgtgaggg gcgagggcga gggcgatgcc accaacggca
6241 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
6301 tgaccaccct gagccacggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc
6361 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
6421 aggacgacgg cacctacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
6481 accgcatcga gctgaagggc gtcgacttca aggaggacgg caacatcctg gggcacaagc
6541 tggagtacaa cttcaacagc cacaacatct atatcatggc cgtcaagcag aagaacggca
6601 tcaaggtgaa cttcaagatc cgccacaacg tggaggacgg cagcgtgcag ctcgccgacc
6661 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac agccactacc
6721 tgagcaccca gtccgtgctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
6781 tggagttccg caccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaacccc
6841 ggggaagttc ctatactttc tagagaatag gaacttcgct agagactgtg ccttctagtt
6901 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
6961 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
7021 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
7081 ggcatgctgg ggatgcggtc tagcgcggcc gccgcgccga tatcgaattc gctagaagat
7141 gggcgggagt cttctgggca ggcttaaagg ctaacctggt gtgtgggcgt tgtcctgcag
7201 gggaattgaa caggtgtaaa attggaggga caagacttcc cacagatttt cggttttgtc
7261 gggaagtttt ttaatagggg caaataagga aaatgggagg ataggtagtc atctggggtt
7321 ttatgcagca aaactacagg ttattattgc ttgtgatccg cctcggagta ttttccatcg
7381 aggtagatta aagacatgct cacccgagtt ttatactctc ctgcttgaga tccttactac
7441 agtatgaaat tacagtgtcg cgagttagac tatgtaagca gaattttaat catttttaaa
7501 gagcccagta cttcatatcc atttctcccg ctccttctgc agccttatca aaaggtattt
7561 tagaacactc attttagccc cattttcatt tattatactg gcttatccaa cccctagaca
7621 gagcattggc attttccctt tcctgatctt agaagtctga tgactcatga aaccagacag
7681 attagttaca tacaccacaa atcgaggctg tagctggggc ctcaacactg cagttctttt
7741 ataactcctt agtacacttt ttgttgatcc tttgccttga tccttaattt tcagtgtcta
7801 tcacctctcc cgtcaggtgg tgttccacat ttgggcctat tctcagtcca gggagtttta
7861 caacaataga tgtattgaga atccaaccta aagcttaact ttccactccc atgaatgcct
7921 ctctcctttt tctccattta taaactgagc tattaaccat taatggtttc caggtggatg
7981 tctcctcccc caatattacc tgatgtatct tacatattgc caggctgata ttttaagaca
8041 ttaaaaggta tatttcatta ttgagccaca tggtattgat tactgcttac taaaattttg
8101 tcattgtaca catctgtaaa aggtggttcc ttttggaatg caaagttcag gtgtttgttg
8161 tctttcctga cctaaggtct tgtgagcttg tattttttct atttaagcag tgctttctct
8221 tggactggct tgactcatgg cattctacac gttattgctg gtctaaatgt gattttgcca
8281 agcttcttca ggacctataa ttttgcttga cttgtagcca aacacaagta aaatgattaa
8341 gcaacaaatg tatttgtgaa gcttggtttt taggttgttg tgttgtgtgt gcttgtgctc
8401 tataataata ctatccaggg gctggagagg tggctcggag ttcaagagca cagactgctc
8461 ttccagaagt cctgagttca attcccagca accacatggt ggctcacaac catctgtaat
8521 gggatctgat gccctcttct ggtgtgtctg aagaccacaa gtgtattcac attaaataaa
8581 taaatcctcc ttcttcttct tttttttttt tttaaagaga atactgtctc cagtagaatt
8641 tactgaagta atgaaatact ttgtgtttgt tccaatatgg tagccaataa tcaaattact
8701 ctttaagcac tggaaatgtt accaaggaac taatttttat ttgaagtgta actgtggaca
8761 gaggagccat aactgcagac ttgtgggata cagaagacca atgcagactt taatgtcttt
8821 tctcttacac taagcaataa agaaataaaa attgaacttc tagtatccta tttgtttaaa
8881 ctgctagctt tacttaactt ttgtgcttca tctatacaaa gctgaaagct aagtctgcag
8941 ccattactaa acatgaaagc aagtaatgat aattttggat ttcaaaaatg tagggccaga
9001 gtttagccag ccagtggtgg tgcttgcctt tatgccttta atcccagcac tctggaggca
9061 gagacaggca gatctctgag tttgagccca gcctggtcta cacatcaagt tctatctagg
9121 atagccagga atacacacag aaaccctgtt ggggaggggg gctctgagat ttcataaaat
9181 tataattgaa gcattcccta atgagccact atggatgtgg ctaaatccgt ctacctttct
9241 gatgagattt gggtattatt ttttctgtct ctgctgttgg ttgggtcttt tgacactgtg
9301 ggctttcttt aaagcctcct tcctgccatg tggtctcttg tttgctacta acttcccatg
9361 gcttaaatgg catggctttt tgccttctaa gggcagctgc tgagatttgc agcctgattt
9421 ccagggtggg gttgggaaat ctttcaaaca ctaaaattgt cctttaattt tttttttaaa
9481 aaatgggtta tataataaac ctcataaaat agttatgagg agtgaggtgg actaatatta
9541 aatgagtccc tcccctataa aagagctatt aaggcttttt gtcttatact taactttttt
9601 tttaaatgtg gtatctttag aaccaagggt cttagagttt tagtatacag aaactgttgc
9661 atcgcttaat cagattttct agtttcaaat ccagagaatc caaattcttc acagccaaag
9721 tcaaattaag aatttctgac ttttaatgtt aatttgctta ctgtgaatat aaaaatgata
9781 gcttttcctg aggcagggtc tcactatgta tctctgcctg atctgcaaca agatatgtag
9841 actaaagttc tgcctgcttt tgtctcctga atactaaggt taaaatgtag taatactttt
9901 ggaacttgca ggtcagattc ttttataggg gacacactaa gggagcttgg gtgatagttg
9961 gtaaaatgtg tttcaagtga tgaaaacttg aattattatc accgcaacct actttttaaa
10021 aaaaaaagcc aggcctgtta gagcatgctt aagggatccc taggacttgc tgagcacaca
10081 agagtagtta cttggcaggc tcctggtgag agcatatttc aaaaaacaag gcagacaacc
10141 aagaaactac agttaaggtt acctgtcttt aaaccatctg catatacaca gggatattaa
10201 aatattccaa ataatatttc attcaagttt tcccccatca aattgggaca tggatttctc
10261 cggtgaatag gcagagttgg aaactaaaca aatgttggtt ttgtgatttg tgaaattgtt
10321 ttcaagtgat agttaaagcc catgagatac agaacaaagc tgctatttcg aggtctcttg
10381 gtttatactc agaagcactt ctttgggttt ccctgcacta tcctgatcat gtgctaggcc
10441 taccttaggc tgattgttgt tcaaataaac ttaagtttcc tgtcaggtga tgtcatatga
10501 tttcatatat caaggcaaaa catgttatat atgttaaaca tttgtactta atgtgaaagt
10561 taggtctttg tgggtttgat ttttaatttt caaaacctga gctaaataag tcatttttac
10621 atgtcttaca tttggtggaa ttgtataatt gtggtttgca ggcaagactc tctgacctag
10681 taaccctacc tatagagcac tttgctgggt cacaagtcta ggagtcaagc atttcacctt
10741 gaagttgaga cgttttgtta gtgtatacta gtttatatgt tggaggacat gtttatccag
10801 aagatattca ggactatttt tgactgggct aaggaattga ttctgattag cactgttagt
10861 gagcattgag tggcctttag gcttgaattg gagtcacttg tatatctcaa ataatgctgg
10921 ccttttttaa aaagcccttg ttctttatca ccctgttttc tacataattt ttgttcaaag
10981 aaatacttgt ttggatctcc ttttgacaac aatagcatgt tttcaagcca tatttttttt
11041 cctttttttt tttttttttg gtttttcgag acagggtttc tctgtatagc cctggctgtc
11101 ctggaactca ctttgtagac caggctggcc tcgaactcag aaatccgcct gcctctgcct
11161 cctgagtgcc gggattaaag gcgtgcacca ccacgcctgg ctaagttgga tattttgtta
11221 tataactata accaatacta actccactgg gtggattttt aattcagtca gtagtcttaa
11281 gtggtcttta ttggcccttc attaaaatct actgttcact ctaacagagg ctgttggtac
11341 tagtggcact taagcaactt cctacggata tactagcaga ttaagggtca gggatagaaa
11401 ctagtctagc gttttgtata cctaccagct ttatactacc ttgttctgat agaaatattt
11461 aggacatcta gcttatcgat ccgtcgacgg tatcgataag cttgatatcg aattctaccg
11521 ggtaggggag gcgcttttcc aaggcagtct gagcatgcgc ttagcagccc cgctggcact
11581 tggcgctaca caagtggcct tggcctcgca cacattccac atccaccggt aggcgccgcc
11641 gccctgcagg cgatcgctac gtacagcttt tgttcccttt agtgagggtt aattgcgcgc
11701 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
11761 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
11821 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
11881 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
11941 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
12001 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
12061 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
12121 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
12181 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
12241 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
12301 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
12361 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
12421 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
12481 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
12541 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
12601 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
12661 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
12721 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
12781 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
12841 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
12901 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
12961 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
13021 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
13081 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
13141 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
13201 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
13261 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
13321 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
13381 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
13441 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
13501 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
13561 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
13621 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
13681 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
13741 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
13801 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
13861 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
13921 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
13981 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
14041 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
14101 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
14161 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
14221 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
14281 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
14341 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
14401 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
14461 gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc gaattggagc
14521 tcc
//
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