| 质粒名称: | pCBh_NLS_hfCas13X(Cas13X_M17YY)_NLS-pA-U6-DR-BpiI-BpiI-DR-pSV40-EGFP-pA-pSV40-mCherry-pA |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 190033 |
| 存储实验室: | Huawei Tong |
|
载体骨架基本信息 |
|
| 载体名称: | piggyBac-CBh |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | 9185 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄青霉素，100μg/mL |
| 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: | humanized hfCas13X |
| 物种来源: | Synthetic |
| 插入大小: | -- |
| 基因突变: | Y672A,Y676A |
| GenBank编号 | -- |
| 启动子: | CBh, SV40, hU6 |
| 标签/融合蛋白: | -- |
| 克隆方法: | Gibson Cloning |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pCBh_NLS_hfCas13X(Cas13X_M17YY)_NLS-pA-U6-DR-BpiI-BpiI-DR-pSV40-EGFP-pA-pSV40-mCherry-pA was a gift from Huawei Tong (Addgene plasmid # 190033 ; http://n2t.net/addgene:190033 ; RRID:Addgene_190033) |
| 参考文献部分: | High-fidelity Cas13 variants for targeted RNA degradation with minimal collateral effects. Tong H, Huang J, Xiao Q, He B, Dong X, Liu Y, Yang X, Han D, Wang Z, Wang X, Ying W, Zhang R, Wei Y, Xu C, Zhou Y, Li Y, Cai M, Wang Q, Xue M, Li G, Fang K, Zhang H, Yang H. Nat Biotechnol. 2022 Aug 11. pii: 10.1038/s41587-022-01419-7. doi: 10.1038/s41587-022-01419-7. 10.1038/s41587-022-01419-7 PubMed 35953673 |
pCBh_NLS_hfCas13X(Cas13X_M17YY)_NLS-pA-U6-DR-BpiI-BpiI-DR-pS
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
_NLS-pA-U6-DR-BpiI-BpiI-DR-pSV40-EGFP-pA-pSV40-mCherry-pA质粒图谱.png)
载体简介
载体序列
LOCUS pCBh_NLS_hfCas13X(Cas13X_M17YY)_NLS-pA-U6-DR-BpiI-BpiI-DR-pSV40-EGFP-pA-pSV40-mCherry-pA 9185 bp ds-DNA circular SYN 12-SEP-2022
DEFINITION vector for encoding a human codon-optimized High-fidelity Cas13X
(hfCas13X) driven by CBh promoter, guide RNAs compatible with Cas13X
driven by hU6, EGFP and mCherry driven by SV40 promoters.
ACCESSION .
VERSION .
KEYWORDS
pCBh_NLS_hfCas13X(Cas13X_M17YY)_NLS-pA-U6-DR-BpiI-BpiI-DR-pSV40-EGFP-pA-pSV40-mCherry-pA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9185)
TITLE High-fidelity Cas13 variants for targeted RNA degradation with
minimal collateral effects
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 9185)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 12, 2022 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..9185
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 25..87
/label=piggyBac right (3') inverted repeat
/note="piggyBac transposon-specific inverted terminal
repeat sequence (ITR)"
enhancer 396..681
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer;
contains an 18-bp deletion relative to the standard CMV
enhancer"
intron 961..1188
/label=hybrid intron
/note="hybrid between chicken beta-actin (CBA) and minute
virus of mice (MMV) introns (Gray et al., 2011)"
CDS 1209..1229
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 3552..3572
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 3582..3716
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
promoter 3723..3963
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 3723..3743
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 3894..3913
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
promoter 4088..4159
/gene="bla"
/label=AmpR promoter
promoter 4161..4518
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 4369..4504
/label=SV40 ori
/note="SV40 origin of replication"
regulatory 4559..4568
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 4565..5281
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(4610..4631)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(4871..4890)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 5218..5239
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 5407..5528
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
promoter 5535..5606
/gene="bla"
/label=AmpR promoter
promoter 5608..5965
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 5816..5951
/label=SV40 ori
/note="SV40 origin of replication"
regulatory 6006..6015
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 6012..6722
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
DDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(6157..6175)
/label=mCherry-R
/note="mCherry, reverse primer"
polyA_signal 6845..6966
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
repeat_region 7323..7357
/label=piggyBac left (5') inverted repeat
/note="piggyBac transposon-specific inverted terminal
repeat sequence (ITR)"
promoter 7388..7492
/gene="bla"
/label=AmpR promoter
CDS 7493..8353
/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 complement(7711..7730)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 8524..9112
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 9013..9032
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tatcccctga ttctctcgag ttaaccctag aaagataatc atattgtgac gtacgttaaa
61 gataatcatg cgtaaaattg acgcatgtgt tttatcgatc tgtatatcga ggtttattta
121 ttaatttgaa tagatattaa gttttattat atttacactt acatactaat aataaattca
181 acaaacaatt tatttatgtt tatttattta ttaaaaaaaa acaaaaactc aaaatttctt
241 ctataaagta acaaaacttt taaacattct ctcttttaca aaaataaact tattttgtac
301 tttaaaaaca gtcatgttgt attataaaat aagtaattag cttaacttat acataataga
361 aacaaattat acttattaat taatctagag gtacccgtta cataacttac ggtaaatggc
421 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caatagtaac gccaataggg
481 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
541 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
601 tggcattgtg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
661 ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc
721 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg
781 atgggggcgg gggggggggg ggggsgsgcg csrggsgggg sggggsgggg sgrggggsgg
841 ggsggggsgr ggsggrgrgg kgsggsggcr gccaatcaga gcggcgcgct ccgaaagttt
901 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
961 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
1021 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
1081 ccgggctgta attagctgag caagaggtaa gggtttaagg gatggttggt tggtggggta
1141 ttaatgttta attacctgga gcacctgcct gaaatcactt tttttcaggt tggaccggtg
1201 ccaccatgcc caagaagaag cggaaggtgg cccaggtgag caagcagacc tccaagaaga
1261 gggagctgag catcgacgag taccagggcg cccggaagtg gtgcttcacc attgccttca
1321 acaaggccct ggtgaaccgg gacaagaacg acggcctgtt cgtggaaagc ctgctgagac
1381 acgagaagta cagcaagcac gactggtacg acgaagatac ccgggccctg atcaagtgca
1441 gcacccaggc cgccaacgcc aaggctgaag ccctgcggaa ctacttcagt cactaccggc
1501 atagccctgg ctgcctgacc ttcaccgccg aggacgaact gcggaccatc atggagagag
1561 cctatgagcg ggccatcttc gagtgcagaa gaagagagac agaggtgatc atcgagtttc
1621 ccagcctgtt cgagggcgac cggatcacca ccgccggcgt ggtgtttttc gtgagctttt
1681 tcgtggaaag aagagtgctg gatcggctgt atggagccgt gtccggcctg aagaagaatg
1741 agggacagta caagctgacc cggaaggccc tgagcatgta ctgcctgaag gacagcagat
1801 tcaccaaggc ctgggataag cgggtgctgc tgttcagaga catcctggcc cagctgggaa
1861 gaatccccgc cgaggcctac gagtactacc acggcgagca gggtgataag aagagagcta
1921 acgacaatga gggcacaaat cccaagcggc acaaggacaa gttcatcgaa tttgcactgc
1981 actacctgga agcccagcac agcgagatct gcttcggcag acgccacatc gtgcgggaag
2041 aggccggcgc cggcgatgag cacaagaagc accggaccaa gggaaaggtg gtggtggact
2101 tcagcaagaa ggacgaggac cagagctact atatctccaa gaacaacgtg atcgtgcgga
2161 tcgacaagaa cgccggccct agaagctacc ggatgggcct gaacgagctg aagtacctcg
2221 tgctgctgag cctgcagggg aagggcgacg atgccatcgc caagctgtac agatacagac
2281 agcacgtgga gaacatcctg gatgtggtga aggtgaccga taaggataac cacgtgttcc
2341 tgccccgctt cgtgctggag cagcacggca tcggcagaaa ggccttcaag cagcggatcg
2401 atggacgggt gaagcacgtg cggggcgtgt gggagaagaa gaaggccgcc accaatgaaa
2461 tgaccctgca cgagaaggcc agagacatcc tgcagtacgt gaacgaaaac tgcacccggt
2521 ccttcaaccc tggcgaatac aacagactgc tggtgtgcct ggtgggcaag gacgtggaga
2581 actttcaggc cggcctgaag cggctgcagc tggccgaaag gatcgatggc cgggtgtact
2641 ccatcttcgc ccagaccagc accatcaatg agatgcacca ggtggtgtgc gaccagatcc
2701 tgaaccggct gtgcagaatc ggcgaccaga agctgtacga ttacgtggga ctgggcaaga
2761 aggacgaaat cgactacaag cagaaggtgg cctggttcaa ggagcacatc agcatccgga
2821 gaggattcct gagaaagaag ttctggtacg atagcaagaa gggattcgca aagctggtgg
2881 aggaacacct ggagtccggc ggcggccagc gcgacgtggg cctggacaag aagtactacc
2941 acatcgacgc catcggcaga ttcgagggcg ccaaccccgc cctgtacgag accctggcca
3001 gagatcggct gtgcctcatg atggcccagt acttcctggg cagcgtgaga aaggaactgg
3061 gcaacaagat tgtgtggagc aacgacagca tcgaactgcc tgtggaaggc tctgtgggaa
3121 atgagaagag catcgtgttc tccgtgtctg actacggcaa gctgtacgtg ctggacgatg
3181 ccgaattcct gggccggatc tgcgaatact tcatgcccca cgaaaagggc aagatccggg
3241 cccacacagt ggccgaaaag ggctttagag catacaacga cctgcagaag aagtgcgtgg
3301 aggccgtgct ggctttcgaa gagaaggtgg tgaaggccaa gaagatgagc gagaaggaag
3361 gcgcccacta catcgacttc cgggagatcc tggcccagac catgtgcaag gaggccgaaa
3421 agaccgcagt gaacaaggtg agacgcgcct tcttccacca ccacctgaag ttcgtgattg
3481 acgagttcgg cctgttcagc gacgtgatga agaagtacgg catcgagaag gaatggaagt
3541 tccctgtcaa gcccaagaag aagcggaagg tgtagaagct taacttgttt attgcagctt
3601 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
3661 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcacta
3721 gtgagggcct atttcccatg attccttcat atttgcatat acgatacaag gctgttagag
3781 agataattgg aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta
3841 gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc
3901 atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag
3961 gacgaaacac cggctggagc agcccccgat ttgtggggtg attacagcgg tcttcgatat
4021 tcaagcgtcg gaagacctgc tggagcagcc cccgatttgt ggggtgatta cagctttttt
4081 tacgcgtcat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
4141 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
4201 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
4261 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
4321 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
4381 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
4441 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
4501 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gctctagagc
4561 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
4621 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
4681 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
4741 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
4801 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
4861 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
4921 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
4981 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
5041 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
5101 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
5161 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
5221 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
5281 gtgaagcggc cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt
5341 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
5401 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
5461 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
5521 gtatcttagt cgaccattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
5581 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
5641 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
5701 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
5761 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
5821 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
5881 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
5941 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgct
6001 ctagagccac catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca
6061 tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg
6121 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg
6181 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct
6241 acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca
6301 agtgggagcg cgtgatgaac ttcgatgacg gcggcgtggt gaccgtgacc caggactcct
6361 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg
6421 acggccccgt aatgcagaag aaaaccatgg gctgggaggc ctcctccgag cggatgtacc
6481 ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc
6541 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg
6601 cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg
6661 aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt
6721 aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
6781 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
6841 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
6901 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
6961 atcttagtcg acggcgcgcc ctagacgaag cagctccagc ctacacaatc gctcaagacg
7021 tgtaatgctt ttattatata ttagtcacga tatctataac aagaaaatat atatataata
7081 agttatcacg taagtagaac atgaaataac aatataatta tcgtatgagt taaatcttaa
7141 aagtcacgta aaagataatc atgcgtcatt ttgactcacg cggtcgttat agttcaaaat
7201 cagtgacact taccgcattg acaagcacgc ctcacgggag ctccaagcgg cgactgagat
7261 gtcctaaatg cacagcgacg gattcgcgct atttagaaag agagagcaat atttcaagaa
7321 tgcatgcgtc aattttacgc agactatctt tctagggtta aggatccgca cttttcgggg
7381 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
7441 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
7501 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
7561 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
7621 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
7681 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
7741 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
7801 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
7861 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
7921 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
7981 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
8041 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
8101 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
8161 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
8221 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
8281 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
8341 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
8401 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
8461 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
8521 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
8581 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
8641 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
8701 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
8761 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
8821 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
8881 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
8941 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
9001 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
9061 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
9121 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
9181 tgcgt
//
