| 质粒名称: | pR26 CAG AsiSI/MluI |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 74286 |
| 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: | 12625bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 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: | |
| 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 CAG AsiSI/MluI 12625 bp ds-DNA circular SYN 23-SEP-2022
DEFINITION Gene targeting vector for the mouse Rosa26 locus, including a CAG
promoter and loxP flanked stop cassette, for cloning into AsiSI or
Mlui.
ACCESSION .
VERSION .
KEYWORDS pR26 CAG AsiSI/MluI
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12625)
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 12625)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 23, 2022 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..12625
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 26..1108
/label=Rosa26 left arm
/note="left homology arm from intron 1 of the mouse Rosa26
gene (Soriano, 1999)"
misc_feature 1131..1234
/label=Ad SA
/note="adenovirus major late transcript splice acceptor
(Friedrich and Soriano, 1991)"
intron 1478..1543
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
polyA_signal 1590..1724
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
protein_bind 1856..1903
/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)."
enhancer 1918..2297
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
intron 2575..3591
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 3515..3537
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
protein_bind 3638..3671
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
CDS 3768..4367
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(3768..3787)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 4264..4284
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
polyA_signal 4597..4731
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
protein_bind 4733..4766
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
primer_bind complement(4857..4873)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4857..4873)
/label=pBluescriptSK
/note="For pBluescript vector"
primer_bind complement(4887..4904)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 4893..5117
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
promoter complement(5161..5178)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 5183..5317
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
misc_feature 5390..9730
/label=Rosa26 right arm
/note="right homology arm from intron 1 of the mouse Rosa26
gene (Soriano, 1999)"
primer_bind complement(9780..9800)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(9780..9798)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(9819..9835)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(9819..9835)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(9832..9854)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 9843..9859
/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(9867..9897)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 9912..9933
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(10050..10067)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(10221..10809)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(10301..10320)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(10980..11840)
/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 11603..11622
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(11841..11945)
/gene="bla"
/label=AmpR promoter
rep_origin 11972..12427
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(12059..12078)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 12269..12290
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 12553..12575
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 12567..12584
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 12568..12584
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 12594..12613
/label=T7
/note="T7 promoter, forward primer"
promoter 12594..12612
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 ccaccgcatt accctgttat ccctagcggc aggccctccg agcgtggtgg agccgttctg
61 tgagacagcc gggtacgagt cgtgacgctg gaaggggcaa gcgggtggtg ggcaggaatg
121 cggtccgccc tgcagcaacc ggagggggag ggagaaggga gcggaaaagt ctccaccgga
181 cgcggccatg gctcgggggg gggggggcag cggaggagcg cttccggccg acgtctcgtc
241 gctgattggc ttcttttcct cccgccgtgt gtgaaaacac aaatggcgtg ttttggttgg
301 cgtaaggcgc ctgtcagtta acggcagccg gagtgcgcag ccgccggcag cctcgctctg
361 cccactgggt ggggcgggag gtaggtgggg tgaggcgagc tggacgtgcg ggcgcggtcg
421 gcctctggcg gggcgggggr ggggagggag ggtcagcgaa agtagctcgc gcgcgagcgg
481 ccgcccaccc tccccttcct ctgggggagt cgttttaccc gccgccggcc gggcctcgtc
541 gtctgattgg ctctcggggc ccagaaaact ggcccttgcc attggctcgt gttcgtgcaa
601 gttgagtcca tccgccggcc agcgggggcg gcgaggaggc gctcccaggt tccggccctc
661 ccctcggccc cgcgccgcag agtctggccg cgcgcccctg cgcaacgtgg caggaagcgc
721 gcgctggggg cggggacggg cagtagggct gagcggctgc ggggcgggtg caagcacgtt
781 tccgacttga gttgcctcaa gaggggcgtg ctgagccaga cctccatcgc gcactccggg
841 gagtggaggg aaggagcgag ggctcagttg ggctgttttg gaggcaggaa gcacttgctc
901 tcccaaagtc gctctgagtt gttatcagta agggagctgc agtggagtag gcggggagaa
961 ggccgcaccc ttctccggag gggggagggg agtgttgcaa tacctttctg ggagttctct
1021 gctgcctcct ggcttctgag gaccgccctg ggcctgggag aatcccttcc ccctcttccc
1081 tcgtgatctg caactccagt ctttctaggc gcgcccgggc tgcagatctg tagggcgcag
1141 tagtccaggg tttccttgat gatgtcatac ttatcctgtc cctttttttt ccacagctcg
1201 cggttgagga caaactcttc gcggtctttc cagtggggat cgacggtatc gataagctgg
1261 ccgctctagt ggccgtacgg gcccacctgc cgggccactt aattaaattt aaatcacaat
1321 tccagctgag cgccggtcgc taccattacc agttggtctg gtgtcaaaaa taataataac
1381 cgggcagggg ggatctgcat ggatctttgt gaaggaacct tacttctgtg gtgtgacata
1441 attggacaaa ctacctacag agatttaaag ctctaaggta aatataaaat ttttaagtgt
1501 ataatgtgtt aaactactga ttctaattgt ttgtgtattt tagattccaa cctatggaac
1561 tgatgaatgg gagcagtggt ggaatgccag atccagacat gataagatac attgatgagt
1621 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
1681 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
1741 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
1801 tctacaaatg tggtatggct gattatgatc tgcggccgtg ctagcgctta agcttgaagt
1861 tcctattccg aagttcctat tctctagaaa gtataggaac ttcggcgcgc cggattcgac
1921 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
1981 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
2041 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
2101 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
2161 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
2221 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
2281 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc
2341 ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg
2401 gggcgggggg gggggggggg cgcgccrggs ggggsggggs ggggsgrggg gsggggsggg
2461 gsgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt
2521 atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc
2581 gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg
2641 ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc
2701 tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg tgaaagcctt
2761 gaggggctcc gggagggccc tttgtgcggg gggagcggct cggggggtgc gtgcgtgtgt
2821 gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga gcgctgcggg
2881 cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc cgggggcggt
2941 gccccgcggt gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg
3001 ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct gcacccccct
3061 ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg gcgtggcgcg
3121 gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg
3181 cctcgggccg gggagggctc gggggagggg cgcggcggcc cccggagcgc cggcggctgt
3241 cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga
3301 cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct
3361 agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
3421 cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg
3481 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
3541 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gatccttaat
3601 taaccgtctt aagcctgtaa caaccggtac agttcgaata acttcgtata gcatacatta
3661 tacgaagtta taagcttgca tgcctgcagg tcggccgcca cgaccggccg gtgccgccac
3721 catcccctga cccacgcccc tgacccctca caaggagacg accttccatg accgagtaca
3781 agcccacggt gcgcctcgcc acccgcgacg acgtcccccg ggccgtacgc accctcgccg
3841 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga cccggaccgc cacatcgagc
3901 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
3961 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
4021 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
4081 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
4141 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
4201 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
4261 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
4321 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg
4381 acccgcagcg cccgaccgaa aggagcgcac gaccccatgg ctccgaccga agccacccgg
4441 ggcggccccg ccgaccccgc acccgccccc gaggcccacc gactctagag gatcataatc
4501 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
4561 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
4621 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
4681 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat ccataacttc
4741 gtatagcata cattatacga agttataccg ggccaccata gtcgcgagta gcttgggcca
4801 gctaggcctt gaccaaagtt cctctggaat tgcgatcgca cgcgtaaatg attgcagatc
4861 cactagttct agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
4921 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
4981 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
5041 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
5101 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcga gcctctagaa
5161 ctatagtgag tcgtattacg tagatccaga catgataaga tacattgatg agtttggaca
5221 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
5281 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
5341 tatgtttcag gttcaggggg aggtgtggga ggttttttaa ttcgcggccc tagaagatgg
5401 gcgggagtct tctgggcagg cttaaaggct aacctggtgt gtgggcgttg tcctgcaggg
5461 gaattgaaca ggtgtaaaat tggagggaca agacttccca cagattttcg gttttgtcgg
5521 gaagtttttt aataggggca aataaggaaa atgggaggat aggtagtcat ctggggtttt
5581 atgcagcaaa actacaggtt attattgctt gtgatccgcc tcggagtatt ttccatcgag
5641 gtagattaaa gacatgctca cccgagtttt atactctcct gcttgagatc cttactacag
5701 tatgaaatta cagtgtcgcg agttagacta tgtaagcaga attttaatca tttttaaaga
5761 gcccagtact tcatatccat ttctcccgct ccttctgcag ccttatcaaa aggtatttta
5821 gaacactcat tttagcccca ttttcattta ttatactggc ttatccaacc cctagacaga
5881 gcattggcat tttccctttc ctgatcttag aagtctgatg actcatgaaa ccagacagat
5941 tagttacata caccacaaat cgaggctgta gctggggcct caacactgca gttcttttat
6001 aactccttag tacacttttt gttgatcctt tgccttgatc cttaattttc agtgtctatc
6061 acctctcccg tcaggtggtg ttccacattt gggcctattc tcagtccagg gagttttaca
6121 acaatagatg tattgagaat ccaacctaaa gcttaacttt ccactcccat gaatgcctct
6181 ctcctttttc tccatttata aactgagcta ttaaccatta atggtttcca ggtggatgtc
6241 tcctccccca atattacctg atgtatctta catattgcca ggctgatatt ttaagacatt
6301 aaaaggtata tttcattatt gagccacatg gtattgatta ctgcttacta aaattttgtc
6361 attgtacaca tctgtaaaag gtggttcctt ttggaatgca aagttcaggt gtttgttgtc
6421 tttcctgacc taaggtcttg tgagcttgta ttttttctat ttaagcagtg ctttctcttg
6481 gactggcttg actcatggca ttctacacgt tattgctggt ctaaatgtga ttttgccaag
6541 cttcttcagg acctataatt ttgcttgact tgtagccaaa cacaagtaaa atgattaagc
6601 aacaaatgta tttgtgaagc ttggttttta ggttgttgtg ttgtgtgtgc ttgtgctcta
6661 taataatact atccaggggc tggagaggtg gctcggagtt caagagcaca gactgctctt
6721 ccagaagtcc tgagttcaat tcccagcaac cacatggtgg ctcacaacca tctgtaatgg
6781 gatctgatgc cctcttctgg tgtgtctgaa gaccacaagt gtattcacat taaataaata
6841 aatcctcctt cttcttcttt tttttttttt taaagagaat actgtctcca gtagaattta
6901 ctgaagtaat gaaatacttt gtgtttgttc caatatggta gccaataatc aaattactct
6961 ttaagcactg gaaatgttac caaggaacta atttttattt gaagtgtaac tgtggacaga
7021 ggagccataa ctgcagactt gtgggataca gaagaccaat gcagacttta atgtcttttc
7081 tcttacacta agcaataaag aaataaaaat tgaacttcta gtatcctatt tgtttaaact
7141 gctagcttta cttaactttt gtgcttcatc tatacaaagc tgaaagctaa gtctgcagcc
7201 attactaaac atgaaagcaa gtaatgataa ttttggattt caaaaatgta gggccagagt
7261 ttagccagcc agtggtggtg cttgccttta tgcctttaat cccagcactc tggaggcaga
7321 gacaggcaga tctctgagtt tgagcccagc ctggtctaca catcaagttc tatctaggat
7381 agccaggaat acacacagaa accctgttgg ggaggggggc tctgagattt cataaaatta
7441 taattgaagc attccctaat gagccactat ggatgtggct aaatccgtct acctttctga
7501 tgagatttgg gtattatttt ttctgtctct gctgttggtt gggtcttttg acactgtggg
7561 ctttctttaa agcctccttc ctgccatgtg gtctcttgtt tgctactaac ttcccatggc
7621 ttaaatggca tggctttttg ccttctaagg gcagctgctg agatttgcag cctgatttcc
7681 agggtggggt tgggaaatct ttcaaacact aaaattgtcc tttaattttt tttttaaaaa
7741 atgggttata taataaacct cataaaatag ttatgaggag tgaggtggac taatattaaa
7801 tgagtccctc ccctataaaa gagctattaa ggctttttgt cttatactta actttttttt
7861 taaatgtggt atctttagaa ccaagggtct tagagtttta gtatacagaa actgttgcat
7921 cgcttaatca gattttctag tttcaaatcc agagaatcca aattcttcac agccaaagtc
7981 aaattaagaa tttctgactt ttaatgttaa tttgcttact gtgaatataa aaatgatagc
8041 ttttcctgag gcagggtctc actatgtatc tctgcctgat ctgcaacaag atatgtagac
8101 taaagttctg cctgcttttg tctcctgaat actaaggtta aaatgtagta atacttttgg
8161 aacttgcagg tcagattctt ttatagggga cacactaagg gagcttgggt gatagttggt
8221 aaaatgtgtt tcaagtgatg aaaacttgaa ttattatcac cgcaacctac tttttaaaaa
8281 aaaaagccag gcctgttaga gcatgcttaa gggatcccta ggacttgctg agcacacaag
8341 agtagttact tggcaggctc ctggtgagag catatttcaa aaaacaaggc agacaaccaa
8401 gaaactacag ttaaggttac ctgtctttaa accatctgca tatacacagg gatattaaaa
8461 tattccaaat aatatttcat tcaagttttc ccccatcaaa ttgggacatg gatttctccg
8521 gtgaataggc agagttggaa actaaacaaa tgttggtttt gtgatttgtg aaattgtttt
8581 caagtgatag ttaaagccca tgagatacag aacaaagctg ctatttcgag gtctcttggt
8641 ttatactcag aagcacttct ttgggtttcc ctgcactatc ctgatcatgt gctaggccta
8701 ccttaggctg attgttgttc aaataaactt aagtttcctg tcaggtgatg tcatatgatt
8761 tcatatatca aggcaaaaca tgttatatat gttaaacatt tgtacttaat gtgaaagtta
8821 ggtctttgtg ggtttgattt ttaattttca aaacctgagc taaataagtc atttttacat
8881 gtcttacatt tggtggaatt gtataattgt ggtttgcagg caagactctc tgacctagta
8941 accctaccta tagagcactt tgctgggtca caagtctagg agtcaagcat ttcaccttga
9001 agttgagacg ttttgttagt gtatactagt ttatatgttg gaggacatgt ttatccagaa
9061 gatattcagg actatttttg actgggctaa ggaattgatt ctgattagca ctgttagtga
9121 gcattgagtg gcctttaggc ttgaattgga gtcacttgta tatctcaaat aatgctggcc
9181 ttttttaaaa agcccttgtt ctttatcacc ctgttttcta cataattttt gttcaaagaa
9241 atacttgttt ggatctcctt ttgacaacaa tagcatgttt tcaagccata ttttttttcc
9301 tttttttttt tttttttggt ttttcgagac agggtttctc tgtatagccc tggctgtcct
9361 ggaactcact ttgtagacca ggctggcctc gaactcagaa atccgcctgc ctctgcctcc
9421 tgagtgccgg gattaaaggc gtgcaccacc acgcctggct aagttggata ttttgttata
9481 taactataac caatactaac tccactgggt ggatttttaa ttcagtcagt agtcttaagt
9541 ggtctttatt ggcccttcat taaaatctac tgttcactct aacagaggct gttggtacta
9601 gtggcactta agcaacttcc tacggatata ctagcagatt aagggtcagg gatagaaact
9661 agtctagcgt tttgtatacc taccagcttt atactacctt gttctgatag aaatatttca
9721 ggacatctag cttatcgata ccgtcgacgg tatcgataag cttgatccag cttttgttcc
9781 ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
9841 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
9901 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
9961 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
10021 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
10081 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
10141 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
10201 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
10261 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
10321 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
10381 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
10441 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
10501 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
10561 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
10621 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
10681 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
10741 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
10801 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
10861 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
10921 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
10981 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
11041 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
11101 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
11161 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
11221 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
11281 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
11341 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
11401 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
11461 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
11521 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
11581 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
11641 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
11701 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
11761 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
11821 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
11881 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
11941 ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg
12001 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
12061 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
12121 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
12181 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
12241 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
12301 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
12361 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
12421 acaatttcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
12481 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
12541 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg
12601 actcactata gggcgaattg gagct
//
