| 质粒名称: | pCBh-NLS_hfCas13d(RfxCas13d_N2V8)_HA_NLS-pA-U6-DR-BpiI-BpiI-pSV40-EGFP-pA-pSV40-mCherry-pA |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 190034 |
| 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: | 9761 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 hfCas13d |
| 物种来源: | Synthetic |
| 插入大小: | -- |
| 基因突变: | A134V,A140V,A141V,A143V |
| 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_hfCas13d(RfxCas13d_N2V8)_HA_NLS-pA-U6-DR-BpiI-BpiI-pSV40-EGFP-pA-pSV40-mCherry-pA was a gift from Huawei Tong (Addgene plasmid # 190034 ; http://n2t.net/addgene:190034 ; RRID:Addgene_190034) |
| 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_hfCas13d(RfxCas13d_N2V8)_HA_NLS-pA-U6-DR-BpiI-BpiI-
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
_HA_NLS-pA-U6-DR-BpiI-BpiI-pSV40-EGFP-pA-pSV40-mCherry-pA质粒图谱.png)
载体简介
载体序列
LOCUS pCBh-NLS_hfCas13d(RfxCas13d_N2V8)_HA_NLS-pA-U6-DR-BpiI-BpiI-pSV40-EGFP-pA-pSV40-mCherry-pA 9761 bp ds-DNA circular SYN 12-SEP-2022
DEFINITION vector for encoding a human codon-optimized High-fidelity RfxCas13d
(hfCas13d) driven by CBh promoter, guide RNAs compatible with
RfxCas13d driven by hU6, EGFP and mCherry driven by SV40 promoters.
ACCESSION .
VERSION .
KEYWORDS
pCBh-NLS_hfCas13d(RfxCas13d_N2V8)_HA_NLS-pA-U6-DR-BpiI-BpiI-pSV40-EGFP-pA-pSV40-mCherry-pA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9761)
TITLE High-fidelity Cas13 variants for targeted RNA degradation with
minimal collateral effects
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 9761)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 12, 2022 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..9761
/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 4146..4172
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 4173..4193
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 4206..4340
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
promoter 4347..4587
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 4347..4367
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 4518..4537
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 4597..4632
/label=RfxCas13d DR36
/note="36-base direct repeat for an RfxCas13d pre-gRNA
(Konermann et al., 2018)"
promoter 4664..4735
/gene="bla"
/label=AmpR promoter
promoter 4737..5094
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 4945..5080
/label=SV40 ori
/note="SV40 origin of replication"
regulatory 5135..5144
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 5141..5857
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(5186..5207)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(5447..5466)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 5794..5815
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 5983..6104
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
promoter 6111..6182
/gene="bla"
/label=AmpR promoter
promoter 6184..6541
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 6392..6527
/label=SV40 ori
/note="SV40 origin of replication"
regulatory 6582..6591
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 6588..7298
/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(6733..6751)
/label=mCherry-R
/note="mCherry, reverse primer"
polyA_signal 7421..7542
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
repeat_region 7899..7933
/label=piggyBac left (5') inverted repeat
/note="piggyBac transposon-specific inverted terminal
repeat sequence (ITR)"
promoter 7964..8068
/gene="bla"
/label=AmpR promoter
CDS 8069..8929
/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(8287..8306)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 9100..9688
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 9589..9608
/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 ggggsgcgcs csrggsgggg sggggsgggg sgrggggsgg
841 ggsggggsgr ggsggrgrgg kgsggsggcr gccaatcaga gcggcgcgct ccgaaagttt
901 ccttttatgg cgrggcggcg 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 taaaaagaaa agaaaggtgg gttctggtat cgagaagaag aagagcttcg
1261 ccaagggcat gggagtgaag agcaccctgg tgtccggctc taaggtgtac atgaccacat
1321 ttgctgaggg aagcgacgcc aggctggaga agatcgtgga gggcgatagc atcagatccg
1381 tgaacgaggg agaggctttc agcgccgaga tggctgacaa gaacgctggc tacaagatcg
1441 gaaacgccaa gttttcccac ccaaagggct acgccgtggt ggctaacaac ccactgtaca
1501 ccggaccagt gcagcaggac atgctgggac tgaaggagac actggagaag aggtacttcg
1561 gcgagtccgc cgacggaaac gataacatct gcatccaggt catccacaac atcctggata
1621 tcgagaagat cctggtggag tacatcacaa acgtggtgta cgtggtgaac aacatctccg
1681 gcctggacaa ggatatcatc ggcttcggaa agttttctac cgtgtacaca tacgacgagt
1741 tcaaggatcc agagcaccac cgggccgctt ttaacaacaa cgacaagctg atcaacgcca
1801 tcaaggctca gtacgacgag ttcgataact ttctggataa ccccaggctg ggctacttcg
1861 gacaggcttt cttttctaag gagggcagaa actacatcat caactacgga aacgagtgtt
1921 acgacatcct ggccctgctg agcggactga ggcactgggt ggtgcacaac aacgaggagg
1981 agtctcggat cagccgcacc tggctgtaca acctggacaa gaacctggat aacgagtaca
2041 tctccacact gaactacctg tacgacagga tcaccaacga gctgacaaac agcttctcca
2101 agaactctgc cgctaacgtg aactacatcg ctgagaccct gggcatcaac ccagctgagt
2161 tcgctgagca gtacttcaga ttttccatca tgaaggagca gaagaacctg ggcttcaaca
2221 tcacaaagct gagagaagtg atgctggaca gaaaggatat gtccgagatc aggaagaacc
2281 acaaggtgtt cgattctatc agaaccaagg tgtacacaat gatggacttt gtgatctaca
2341 ggtactacat cgaggaggat gccaaggtgg ccgctgccaa caagagcctg cccgacaacg
2401 agaagtctct gagcgagaag gatatcttcg tgatcaacct gagaggctcc tttaacgacg
2461 atcagaagga cgctctgtac tacgatgagg ccaacaggat ctggagaaag ctggagaaca
2521 tcatgcacaa catcaaggag ttccggggaa acaagacccg cgagtacaag aagaaggacg
2581 ctccaaggct gcctaggatc ctgcctgctg gaagggacgt gagcgccttc agcaagctga
2641 tgtacgccct gacaatgttt ctggacggaa aggagatcaa cgatctgctg accacactga
2701 tcaacaagtt cgacaacatc cagtcttttc tgaaagtgat gcctctgatc ggcgtgaacg
2761 ctaagttcgt ggaggagtac gccttcttta aggacagcgc caagatcgct gatgagctgc
2821 ggctgatcaa gtcctttgcc aggatgggag agccaatcgc tgacgctagg agagctatgt
2881 acatcgatgc catccggatc ctgggaacca acctgtctta cgacgagctg aaggctctgg
2941 ccgacacctt cagcctggat gagaacggca acaagctgaa gaagggcaag cacggaatgc
3001 gcaacttcat catcaacaac gtgatcagca acaagcggtt tcactacctg atcagatacg
3061 gcgacccagc tcacctgcac gagatcgcta agaacgaggc cgtggtgaag ttcgtgctgg
3121 gacggatcgc cgatatccag aagaagcagg gccagaacgg aaagaaccag atcgaccgct
3181 actacgagac ctgcatcggc aaggataagg gaaagtccgt gtctgagaag gtggacgctc
3241 tgaccaagat catcacaggc atgaactacg accagttcga taagaagaga tctgtgatcg
3301 aggacaccgg aagggagaac gccgagagag agaagtttaa gaagatcatc agcctgtacc
3361 tgacagtgat ctaccacatc ctgaagaaca tcgtgaacat caacgctaga tacgtgatcg
3421 gcttccactg cgtggagcgc gatgcccagc tgtacaagga gaagggatac gacatcaacc
3481 tgaagaagct ggaggagaag ggctttagct ccgtgaccaa gctgtgcgct ggaatcgacg
3541 agacagcccc cgacaagagg aaggatgtgg agaaggagat ggccgagaga gctaaggaga
3601 gcatcgactc cctggagtct gctaacccta agctgtacgc caactacatc aagtactccg
3661 atgagaagaa ggccgaggag ttcaccaggc agatcaacag agagaaggcc aagaccgctc
3721 tgaacgccta cctgaggaac acaaagtgga acgtgatcat ccgggaggac ctgctgcgca
3781 tcgataacaa gacctgtaca ctgttccgga acaaggctgt gcacctggag gtggctcgct
3841 acgtgcacgc ctacatcaac gacatcgccg aggtgaactc ctactttcag ctgtaccact
3901 acatcatgca gaggatcatc atgaacgaga gatacgagaa gtctagcggc aaggtgtctg
3961 agtacttcga cgccgtgaac gatgagaaga agtacaacga tagactgctg aagctgctgt
4021 gcgtgccttt cggatactgt atcccacggt ttaagaacct gagcatcgag gccctgttcg
4081 accgcaacga ggctgccaag tttgataagg agaagaagaa ggtgagcggc aactccggtt
4141 ctggttaccc atacgacgta ccagattacg ctcccaagaa gaagcgcaag gtgggataaa
4201 agcttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
4261 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
4321 tatcttatca tgtctggatc actagtgagg gcctatttcc catgattcct tcatatttgc
4381 atatacgata caaggctgtt agagagataa ttggaattaa tttgactgta aacacaaaga
4441 tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa
4501 aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct
4561 tggctttata tatcttgtgg aaaggacgaa acaccgcaag taaaccccta ccaactggtc
4621 ggggtttgaa acgggtcttc gagaagacct tttttttacg cgtcattcaa atatgtatcc
4681 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
4741 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
4801 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
4861 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
4921 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
4981 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
5041 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
5101 tcaagagaca ggatgaggat cgtttcgctc tagagccacc atggtgagca agggcgagga
5161 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
5221 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
5281 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
5341 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
5401 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
5461 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
5521 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
5581 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
5641 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
5701 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
5761 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
5821 cgccgggatc actctcggca tggacgagct gtacaagtga agcggccgcg actctagatc
5881 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
5941 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
6001 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
6061 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttagtcgac cattcaaata
6121 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
6181 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
6241 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
6301 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
6361 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
6421 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
6481 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
6541 agatcgatca agagacagga tgaggatcgt ttcgctctag agccaccatg gtgagcaagg
6601 gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg
6661 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
6721 gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca
6781 tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc
6841 ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
6901 atgacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct
6961 acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaaa
7021 ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg
7081 agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca
7141 cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg
7201 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc
7261 gccactccac cggcggcatg gacgagctgt acaagtaaag cggccgcgac tctagatcat
7321 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
7381 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
7441 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
7501 gcattctagt tgtggtttgt ccaaactcat caatgtatct tagtcgacgg cgcgccctag
7561 acgaagcagc tccagcctac acaatcgctc aagacgtgta atgcttttat tatatattag
7621 tcacgatatc tataacaaga aaatatatat ataataagtt atcacgtaag tagaacatga
7681 aataacaata taattatcgt atgagttaaa tcttaaaagt cacgtaaaag ataatcatgc
7741 gtcattttga ctcacgcggt cgttatagtt caaaatcagt gacacttacc gcattgacaa
7801 gcacgcctca cgggagctcc aagcggcgac tgagatgtcc taaatgcaca gcgacggatt
7861 cgcgctattt agaaagagag agcaatattt caagaatgca tgcgtcaatt ttacgcagac
7921 tatctttcta gggttaagga tccgcacttt tcggggaaat gtgcgcggaa cccctatttg
7981 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
8041 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
8101 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
8161 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
8221 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
8281 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
8341 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
8401 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
8461 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
8521 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
8581 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
8641 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
8701 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
8761 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
8821 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
8881 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
8941 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
9001 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
9061 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
9121 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
9181 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
9241 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
9301 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
9361 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
9421 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
9481 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
9541 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
9601 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
9661 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
9721 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg t
//
