| 质粒名称: | SECURE BE3(R33A/K34A)-P2A-EGFP (pJUL1410) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 123615 |
| 存储实验室: | Keith Joung |
|
载体骨架基本信息 |
|
| 载体名称: | CAG |
| 载体出品公司: | pSQT817 (Addgene #53373) |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4843 bp |
| 完整质粒大小: | 10768 bp |
| 载体类型: | 哺乳细胞表达 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | BE3(R33A/K34A)-P2A-EGFP,rAPOBEC1(R33A/K34A)-XTEN-hSpCas9n(D10A)-UGI-NLS(SV40)-P2A-EGFP |
| 物种来源: | R. norvegicus (rat), Synthetic; Streptococcus pyogenes, Bacillus subtilis bacteriophage PBS1 |
| 插入大小: | 5925 bp |
| 基因突变: | R33A/K34A in rAPOBEC1, D10A in SpCas9 |
| 启动子 | CAG |
| 克隆方法: | Gibson克隆 |
| 5' 测序引物 | CTCTGCTAACCATGTTCATGCC |
| 3' 测序引物 | CAGAGGGAAAAAGATCTCAGTGG |
|
文献引用方法 |
|
| 材料和方法部分: | SECURE BE3(R33A/K34A)-P2A-EGFP (pJUL1410) was a gift from Keith Joung (Addgene plasmid # 123615 ; http://n2t.net/addgene:123615 ; RRID:Addgene_123615) |
| 参考文献部分: | Transcriptome-wide off-target RNA editing induced by CRISPR-guided DNA base editors. Grunewald J, Zhou R, Garcia SP, Iyer S, Lareau CA, Aryee MJ, Joung JK. Nature. 2019 Apr 17. pii: 10.1038/s41586-019-1161-z. doi: 10.1038/s41586-019-1161-z. 10.1038/s41586-019-1161-z PubMed 30995674 |
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
-P2A-EGFP (pJUL1410)质粒图谱.png)
载体简介
载体序列
LOCUS SECURE BE3(R33A/K34A)-P2A-EGFP (pJUL1410) 10768 bp ds-DNA circular SYN 26-MAR-2019
DEFINITION CAG promoter expression plasmid for
rAPOBEC1(R33A/K34A)-XTEN-hSpCas9n(D10A)-UGI-NLS(SV40)-P2A-EGFP
(SECURE variant BE3-R33A/K34A)..
ACCESSION .
VERSION .
KEYWORDS SECURE BE3(R33A/K34A)-P2A-EGFP (pJUL1410)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10768)
TITLE Expression plasmids for Base Editors
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 10768)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Mar 26, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..10768
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(87..106)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_site 152..207
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal"
primer_bind complement(206..225)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(568..584)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(568..584)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(581..603)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 592..608
/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(616..646)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 661..682
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 741..936
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 787..922
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 849..868
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 942..1076
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(979..998)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1033..1052
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(1144..1161)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1315..1903)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1395..1414)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2074..2934)
/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 2697..2716
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2935..3039)
/gene="bla"
/label=AmpR promoter
enhancer 3070..3449
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 3451..3728
/label=chicken beta-actin promoter
intron 3729..4745
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 4669..4691
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 4753..4772
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
CDS 5579..9679
/codon_start=1
/product="nickase mutant of the Cas9 endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system"
/label=Cas9(D10A)
/note="generates RNA-guided single strand nicks in DNA due
to the D10A mutation in the RuvC catalytic domain"
/translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 9692..9940
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
CDS 9953..9973
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 9992..10048
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS 10049..10768
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(10094..10115)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(10355..10374)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 10702..10723
/label=EGFP-C
/note="EGFP, forward primer"
ORIGIN
1 agcggccgca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat
61 gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc
121 atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata
181 gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta
241 aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc
301 atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat
361 tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg
421 ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc
481 ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac
541 ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
601 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
661 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
721 cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact
781 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
841 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
901 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt
961 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
1021 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct
1081 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
1141 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
1201 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
1261 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
1321 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
1381 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
1441 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
1501 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
1561 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
1621 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
1681 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
1741 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
1801 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
1861 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
1921 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
1981 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
2041 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
2101 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
2161 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
2221 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
2281 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
2341 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
2401 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
2461 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
2521 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
2581 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
2641 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
2701 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
2761 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
2821 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
2881 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
2941 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
3001 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
3061 acctgggtcg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
3121 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
3181 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
3241 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
3301 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
3361 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
3421 tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc
3481 tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt
3541 gtgcagcgat gggggcgggg gggggggggg ggcgcgcgcc rggggggggg gggggggggg
3601 gggggggggg gggggsgggg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc
3661 gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc
3721 ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc
3781 gccgcccgcc ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc
3841 cttctcctcc gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc
3901 tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggg
3961 gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct
4021 gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg
4081 cggccggggg cggtgccccg cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc
4141 ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc
4201 ccctgcaccc ccctccccga gttgctgagc acggcccggc ttcgggtgcg gggctccgta
4261 cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg
4321 gcggggcggg gccgcctcgg gccggggagg gctcggggga ggggcgcggc ggcccccgga
4381 gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg taatcgtgcg
4441 agagggcgca gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc
4501 gccgcacccc ctctagcggg cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg
4561 gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc cagcctcggg
4621 gctgtccgcg gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct
4681 ggcgtgtgac cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc
4741 tacagctcct gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagaattct
4801 gcagtcgacg gtaccgcggg cccgggatcc accggtcgcc accatgagct cagagactgg
4861 cccagtggct gtggacccca cattgagacg gcggatcgag ccccatgagt ttgaggtatt
4921 cttcgatccg agagagctcg ctgctgagac ctgcctgctt tacgaaatta attggggggg
4981 ccggcactcc atttggcgac atacatcaca gaacactaac aagcacgtcg aagtcaactt
5041 catcgagaag ttcacgacag aaagatattt ctgtccgaac acaaggtgca gcattacctg
5101 gtttctcagc tggagcccat gcggcgaatg tagtagggcc atcactgaat tcctgtcaag
5161 gtatccccac gtcactctgt ttatttacat cgcaaggctg taccaccacg ctgacccccg
5221 caatcgacaa ggcctgcggg atttgatctc ttcaggtgtg actatccaaa ttatgactga
5281 gcaggagtca ggatactgct ggagaaactt tgtgaattat agcccgagta atgaagccca
5341 ctggcctagg tatccccatc tgtgggtacg actgtacgtt cttgaactgt actgcatcat
5401 actgggcctg cctccttgtc tcaacattct gagaaggaag cagccacagc tgacattctt
5461 taccatcgct cttcagtctt gtcattacca gcgactgccc ccacacattc tctgggccac
5521 cgggttgaaa agcggcagcg agactcccgg gacctcagag tccgccacac ccgaaagtga
5581 taaaaagtat tctattggtt tagccatcgg cactaattcc gttggatggg ctgtcataac
5641 cgatgaatac aaagtacctt caaagaaatt taaggtgttg gggaacacag accgtcattc
5701 gattaaaaag aatcttatcg gtgccctcct attcgatagt ggcgaaacgg cagaggcgac
5761 tcgcctgaaa cgaaccgctc ggagaaggta tacacgtcgc aagaaccgaa tatgttactt
5821 acaagaaatt tttagcaatg agatggccaa agttgacgat tctttctttc accgtttgga
5881 agagtccttc cttgtcgaag aggacaagaa acatgaacgg caccccatct ttggaaacat
5941 agtagatgag gtggcatatc atgaaaagta cccaacgatt tatcacctca gaaaaaagct
6001 agttgactca actgataaag cggacctgag gttaatctac ttggctcttg cccatatgat
6061 aaagttccgt gggcactttc tcattgaggg tgatctaaat ccggacaact cggatgtcga
6121 caaactgttc atccagttag tacaaaccta taatcagttg tttgaagaga accctataaa
6181 tgcaagtggc gtggatgcga aggctattct tagcgcccgc ctctctaaat cccgacggct
6241 agaaaacctg atcgcacaat tacccggaga gaagaaaaat gggttgttcg gtaaccttat
6301 agcgctctca ctaggcctga caccaaattt taagtcgaac ttcgacttag ctgaagatgc
6361 caaattgcag cttagtaagg acacgtacga tgacgatctc gacaatctac tggcacaaat
6421 tggagatcag tatgcggact tatttttggc tgccaaaaac cttagcgatg caatcctcct
6481 atctgacata ctgagagtta atactgagat taccaaggcg ccgttatccg cttcaatgat
6541 caaaaggtac gatgaacatc accaagactt gacacttctc aaggccctag tccgtcagca
6601 actgcctgag aaatataagg aaatattctt tgatcagtcg aaaaacgggt acgcaggtta
6661 tattgacggc ggagcgagtc aagaggaatt ctacaagttt atcaaaccca tattagagaa
6721 gatggatggg acggaagagt tgcttgtaaa actcaatcgc gaagatctac tgcgaaagca
6781 gcggactttc gacaacggta gcattccaca tcaaatccac ttaggcgaat tgcatgctat
6841 acttagaagg caggaggatt tttatccgtt cctcaaagac aatcgtgaaa agattgagaa
6901 aatcctaacc tttcgcatac cttactatgt gggacccctg gcccgaggga actctcggtt
6961 cgcatggatg acaagaaagt ccgaagaaac gattactcca tggaattttg aggaagttgt
7021 cgataaaggt gcgtcagctc aatcgttcat cgagaggatg accaactttg acaagaattt
7081 accgaacgaa aaagtattgc ctaagcacag tttactttac gagtatttca cagtgtacaa
7141 tgaactcacg aaagttaagt atgtcactga gggcatgcgt aaacccgcct ttctaagcgg
7201 agaacagaag aaagcaatag tagatctgtt attcaagacc aaccgcaaag tgacagttaa
7261 gcaattgaaa gaggactact ttaagaaaat tgaatgcttc gattctgtcg agatctccgg
7321 ggtagaagat cgatttaatg cgtcacttgg tacgtatcat gacctcctaa agataattaa
7381 agataaggac ttcctggata acgaagagaa tgaagatatc ttagaagata tagtgttgac
7441 tcttaccctc tttgaagatc gggaaatgat tgaggaaaga ctaaaaacat acgctcacct
7501 gttcgacgat aaggttatga aacagttaaa gaggcgtcgc tatacgggct ggggacgatt
7561 gtcgcggaaa cttatcaacg ggataagaga caagcaaagt ggtaaaacta ttctcgattt
7621 tctaaagagc gacggcttcg ccaataggaa ctttatgcag ctgatccatg atgactcttt
7681 aaccttcaaa gaggatatac aaaaggcaca ggtttccgga caaggggact cattgcacga
7741 acatattgcg aatcttgctg gttcgccagc catcaaaaag ggcatactcc agacagtcaa
7801 agtagtggat gagctagtta aggtcatggg acgtcacaaa ccggaaaaca ttgtaatcga
7861 gatggcacgc gaaaatcaaa cgactcagaa ggggcaaaaa aacagtcgag agcggatgaa
7921 gagaatagaa gagggtatta aagaactggg cagccagatc ttaaaggagc atcctgtgga
7981 aaatacccaa ttgcagaacg agaaacttta cctctattac ctacaaaatg gaagggacat
8041 gtatgttgat caggaactgg acataaaccg tttatctgat tacgacgtcg atcacattgt
8101 accccaatcc tttttgaagg acgattcaat cgacaataaa gtgcttacac gctcggataa
8161 gaaccgaggg aaaagtgaca atgttccaag cgaggaagtc gtaaagaaaa tgaagaacta
8221 ttggcggcag ctcctaaatg cgaaactgat aacgcaaaga aagttcgata acttaactaa
8281 agctgagagg ggtggcttgt ctgaacttga caaggccgga tttattaaac gtcagctcgt
8341 ggaaacccgc caaatcacaa agcatgttgc acagatacta gattcccgaa tgaatacgaa
8401 atacgacgag aacgataagc tgattcggga agtcaaagta atcactttaa agtcaaaatt
8461 ggtgtcggac ttcagaaagg attttcaatt ctataaagtt agggagataa ataactacca
8521 ccatgcgcac gacgcttatc ttaatgccgt cgtagggacc gcactcatta agaaataccc
8581 gaagctagaa agtgagtttg tgtatggtga ttacaaagtt tatgacgtcc gtaagatgat
8641 cgcgaaaagc gaacaggaga taggcaaggc tacagccaaa tacttctttt attctaacat
8701 tatgaatttc tttaagacgg aaatcactct ggcaaacgga gagatacgca aacgaccttt
8761 aattgaaacc aatggggaga caggtgaaat cgtatgggat aagggccggg acttcgcgac
8821 ggtgagaaaa gttttgtcca tgccccaagt caacatagta aagaaaactg aggtgcagac
8881 cggagggttt tcaaaggaat cgattcttcc aaaaaggaat agtgataagc tcatcgctcg
8941 taaaaaggac tgggacccga aaaagtacgg tggcttcgat agccctacag ttgcctattc
9001 tgtcctagta gtggcaaaag ttgagaaggg aaaatccaag aaactgaagt cagtcaaaga
9061 attattgggg ataacgatta tggagcgctc gtcttttgaa aagaacccca tcgacttcct
9121 tgaggcgaaa ggttacaagg aagtaaaaaa ggatctcata attaaactac caaagtatag
9181 tctgtttgag ttagaaaatg gccgaaaacg gatgttggct agcgccggag agcttcaaaa
9241 ggggaacgaa ctcgcactac cgtctaaata cgtgaatttc ctgtatttag cgtcccatta
9301 cgagaagttg aaaggttcac ctgaagataa cgaacagaag caactttttg ttgagcagca
9361 caaacattat ctcgacgaaa tcatagagca aatttcggaa ttcagtaaga gagtcatcct
9421 agctgatgcc aatctggaca aagtattaag cgcatacaac aagcacaggg ataaacccat
9481 acgtgagcag gcggaaaata ttatccattt gtttactctt accaacctcg gcgctccagc
9541 cgcattcaag tattttgaca caacgataga tcgcaaacga tacacttcta ccaaggaggt
9601 gctagacgcg acactgattc accaatccat cacgggatta tatgaaactc ggatagattt
9661 gtcacagctt gggggtgact ctggtggttc tactaatctg tcagatatta ttgaaaagga
9721 gaccggtaag caactggtta tccaggaatc catcctcatg ctcccagagg aggtggaaga
9781 agtcattggg aacaagccgg aaagcgatat actcgtgcac accgcctacg acgagagcac
9841 cgacgagaat gtcatgcttc tgactagcga cgcccctgaa tacaagcctt gggctctggt
9901 catacaggat agcaacggtg agaacaagat taagatgctc tctggtggtt ctcccaagaa
9961 gaagaggaaa gtcggaggcg gtggaagcgg cgcaacaaac ttctctctgc tgaaacaagc
10021 cggagatgtc gaagagaatc ctggaccgat ggtgagcaag ggcgaggagc tgttcaccgg
10081 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
10141 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
10201 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
10261 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
10321 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
10381 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
10441 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
10501 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
10561 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
10621 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
10681 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
10741 tctcggcatg gacgagctgt acaagtga
//
