| 质粒名称: | pCAG-enAsCas12a-HF1(E174R/N282A/S542R/K548R)-NLS(nuc)-3xHA (AAS1815) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 107942 |
| 存储实验室: | Keith Joung, Benjamin Kleinstiver |
|
载体骨架基本信息 |
|
| 载体名称: | pCAG-CFP |
| 载体出品公司: | Connie Cepko lab (Addgene plasmid #11179) |
| 载体抗性: | 氨苄青霉素 |
| 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: | human codon optimized enAsCas12a-HF1 (E174R/N282A/S542R/K548R),AAS1815,enhanced, high-fidelity version of Cas12a from Acidaminococcus sp. BV3L6 |
| 物种来源: | Acidaminococcus sp. BV3L6 |
| 基因突变: | E174R, S542R, K548R, and N282A |
| 启动子 | CAG |
| 标签/融合蛋白: |
NLS (nucleoplasmin)（插入物上的C端）
3x HA（插入件上的C端子）
|
| 克隆方法: | Gibson克隆 |
| 5' 测序引物 | pCAG-F (5'-GCAACGTGCTGGTTATTGTG-3') |
| 3' 测序引物 | Bglob-pA-R (5'-ttttggcagagggaaaaaga-3') |
|
文献引用方法 |
|
| 材料和方法部分: | pCAG-enAsCas12a-HF1(E174R/N282A/S542R/K548R)-NLS(nuc)-3xHA (AAS1815) was a gift from Keith Joung & Benjamin Kleinstiver (Addgene plasmid # 107942 ; http://n2t.net/addgene:107942 ; RRID:Addgene_107942) |
| 参考文献部分: | Engineered CRISPR-Cas12a variants with increased activities and improved targeting ranges for gene, epigenetic and base editing. Kleinstiver BP, Sousa AA, Walton RT, Tak YE, Hsu JY, Clement K, Welch MM, Horng JE, Malagon-Lopez J, Scarfo I, Maus MV, Pinello L, Aryee MJ, Joung JK. Nat Biotechnol. 2019 Feb 11. pii: 10.1038/s41587-018-0011-0. doi: 10.1038/s41587-018-0011-0. 10.1038/s41587-018-0011-0 PubMed 30742127 |
pCAG-enAsCas12a-HF1(E174R/N282A/S542R/K548R)-NLS(nuc)-3xHA (
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(nuc)-3xHA (AAS1815)质粒图谱.png)
载体简介
载体序列
LOCUS pCAG-enAsCas12a-HF1(E174R/N282A/S542R/K548R)-NLS(nuc)-3xHA (AAS1815) 8893 bp ds-DNA circular SYN 07-AUG-2018
DEFINITION Mammalian expression plasmid for human codon optimized
enAsCas12a-HF1 (enhanced high-fidelity AsCas12a) encoding
E174R/N282A/S542R/K548R substitutions.
ACCESSION .
VERSION .
KEYWORDS pCAG-enAsCas12a-HF1(E174R/N282A/S542R/K548R)-NLS(nuc)-3xHA (AAS1815)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8893)
AUTHORS Keith Joung
TITLE Joung lab Cas12a plasmids
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 8893)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 7, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8893
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 5..384
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 387..662
/label=chicken beta-actin promoter
intron 663..1671
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1595..1617
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1679..1698
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
CDS 1770..5690
/codon_start=1
/product="CRISPR-associated protein from Acidaminococcus
sp. BV3L6 (Zetsche et al., 2015)"
/label=AsCpf1
/translation="MTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARND
HYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRN
AIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFD
KFTTYFSGFYRNRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHF
ENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVL
ALAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLL
RNENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTG
KITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTL
KKQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKA
RNYATKKPYSVEKFKLNFQMPTLARGWDVNREKNNGAILFVKNGLYYLGIMPKQKGRYK
ALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEP
LEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSID
LSSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKG
HHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLK
DQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFF
HVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQR
SLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAV
VVLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQ
FTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYD
VKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIEN
HRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQM
RNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKES
KDLKLQNGISNQDWLAYIQELRN"
regulatory 5410..5419
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 5691..5738
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 5745..5825
/codon_start=1
/product="three tandem HA epitope tags"
/label=3xHA
/translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
primer_bind complement(5915..5934)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_site 5980..6035
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal"
primer_bind complement(6034..6053)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(6396..6412)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6396..6412)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6409..6431)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6420..6436
/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(6444..6474)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6489..6510
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 6569..6764
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 6615..6750
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 6677..6696
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 6770..6904
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6807..6826)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 6861..6880
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(6972..6989)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(7143..7731)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7223..7242)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7902..8762)
/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 8525..8544
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8763..8867)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
181 gggactttcc attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta
241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
361 gtattagtca tcgctattac catgggtcga ggtgagcccc acgttctgct tcactctccc
421 catctccccc ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc
481 agcgatgggg gcgggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc
541 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
661 cgggagtcgc tgcgttgcct tcgccccgtg ccccgctccg cgccgcctcg cgccgcccgc
721 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
781 cgggctgtaa ttagcgcttg gtttaatgac ggctcgtttc ttttctgtgg ctgcgtgaaa
841 gccttaaagg gctccgggag ggccctttgt gcggggggga gcggctcggg gggtgcgtgc
901 gtgtgtgtgt gcgtggggag cgccgcgtgc ggcccgcgct gcccggcggc tgtgagcgct
961 gcgggcgcgg cgcggggctt tgtgcgctcc gcgtgtgcgc gaggggagcg cggccggggg
1021 cggtgccccg cggtgcgggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc
1081 gtgggggggt gagcaggggg tgtgggcgcg gcggtcgggc tgtaaccccc ccctgcaccc
1141 ccctccccga gttgctgagc acggcccggc ttcgggtgcg gggctccgtg cggggcgtgg
1201 cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg
1261 gccgcctcgg gccggggagg gctcggggga ggggcgcggc ggccccggag cgccggcggc
1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
1381 ggacttcctt tgtcccaaat ctggcggagc cgaaatctgg gaggcgccgc cgcaccccct
1441 ctagcgggcg cgggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct
1501 tcgtgcgtcg ccgcgccgcc gtccccttct ccatctccag cctcggggct gccgcagggg
1561 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
1621 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
1681 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattctgcag tcgacggtac
1741 cgcgggcccg ggatccaccg gtcgccacca tgacacagtt cgagggcttt accaacctgt
1801 atcaggtgag caagacactg cggtttgagc tgatcccaca gggcaagacc ctgaagcaca
1861 tccaggagca gggcttcatc gaggaggaca aggcccgcaa tgatcactac aaggagctga
1921 agcccatcat cgatcggatc tacaagacct atgccgacca gtgcctgcag ctggtgcagc
1981 tggattggga gaacctgagc gccgccatcg actcctatag aaaggagaaa accgaggaga
2041 caaggaacgc cctgatcgag gagcaggcca catatcgcaa tgccatccac gactacttca
2101 tcggccggac agacaacctg accgatgcca tcaataagag acacgccgag atctacaagg
2161 gcctgttcaa ggccgagctg tttaatggca aggtgctgaa gcagctgggc accgtgacca
2221 caaccgagca cgagaacgcc ctgctgcgga gcttcgacaa gtttacaacc tacttctccg
2281 gcttttatag aaacaggaag aacgtgttca gcgccgagga tatcagcaca gccatcccac
2341 accgcatcgt gcaggacaac ttccccaagt ttaaggagaa ttgtcacatc ttcacacgcc
2401 tgatcaccgc cgtgcccagc ctgcgggagc actttgagaa cgtgaagaag gccatcggca
2461 tcttcgtgag cacctccatc gaggaggtgt tttccttccc tttttataac cagctgctga
2521 cacagaccca gatcgacctg tataaccagc tgctgggagg aatctctcgg gaggcaggca
2581 ccgagaagat caagggcctg aacgaggtgc tggccctggc catccagaag aatgatgaga
2641 cagcccacat catcgcctcc ctgccacaca gattcatccc cctgtttaag cagatcctgt
2701 ccgataggaa caccctgtct ttcatcctgg aggagtttaa gagcgacgag gaagtgatcc
2761 agtccttctg caagtacaag acactgctga gaaacgagaa cgtgctggag acagccgagg
2821 ccctgtttaa cgagctgaac agcatcgacc tgacacacat cttcatcagc cacaagaagc
2881 tggagacaat cagcagcgcc ctgtgcgacc actgggatac actgaggaat gccctgtatg
2941 agcggagaat ctccgagctg acaggcaaga tcaccaagtc tgccaaggag aaggtgcagc
3001 gcagcctgaa gcacgaggat atcaacctgc aggagatcat ctctgccgca ggcaaggagc
3061 tgagcgaggc cttcaagcag aaaaccagcg agatcctgtc ccacgcacac gccgccctgg
3121 atcagccact gcctacaacc ctgaagaagc aggaggagaa ggagatcctg aagtctcagc
3181 tggacagcct gctgggcctg taccacctgc tggactggtt tgccgtggat gagtccaacg
3241 aggtggaccc cgagttctct gcccggctga ccggcatcaa gctggagatg gagccttctc
3301 tgagcttcta caacaaggcc agaaattatg ccaccaagaa gccctactcc gtggagaagt
3361 tcaagctgaa ctttcagatg cctacactgg ccagaggctg ggacgtgaat agagagaaga
3421 acaatggcgc catcctgttt gtgaagaacg gcctgtacta tctgggcatc atgccaaagc
3481 agaagggcag gtataaggcc ctgagcttcg agcccacaga gaaaaccagc gagggctttg
3541 ataagatgta ctatgactac ttccctgatg ccgccaagat gatcccaaag tgcagcaccc
3601 agctgaaggc cgtgacagcc cactttcaga cccacacaac ccccatcctg ctgtccaaca
3661 atttcatcga gcctctggag atcacaaagg agatctacga cctgaacaat cctgagaagg
3721 agccaaagaa gtttcagaca gcctacgcca agaaaaccgg cgaccagaag ggctacagag
3781 aggccctgtg caagtggatc gacttcacaa gggattttct gtccaagtat accaagacaa
3841 cctctatcga tctgtctagc ctgcggccat cctctcagta taaggacctg ggcgagtact
3901 atgccgagct gaatcccctg ctgtaccaca tcagcttcca gagaatcgcc gagaaggaga
3961 tcatggatgc cgtggagaca ggcaagctgt acctgttcca gatctataac aaggactttg
4021 ccaagggcca ccacggcaag cctaatctgc acacactgta ttggaccggc ctgttttctc
4081 cagagaacct ggccaagaca agcatcaagc tgaatggcca ggccgagctg ttctaccgcc
4141 ctaagtccag gatgaagagg atggcacacc ggctgggaga gaagatgctg aacaagaagc
4201 tgaaggatca gaaaacccca atccccgaca ccctgtacca ggagctgtac gactatgtga
4261 atcacagact gtcccacgac ctgtctgatg aggccagggc cctgctgccc aacgtgatca
4321 ccaaggaggt gtctcacgag atcatcaagg ataggcgctt taccagcgac aagttctttt
4381 tccacgtgcc tatcacactg aactatcagg ccgccaattc cccatctaag ttcaaccaga
4441 gggtgaatgc ctacctgaag gagcaccccg agacacctat catcggcatc gatcggggcg
4501 agagaaacct gatctatatc acagtgatcg actccaccgg caagatcctg gagcagcgga
4561 gcctgaacac catccagcag tttgattacc agaagaagct ggacaacagg gagaaggaga
4621 gggtggcagc aaggcaggcc tggtctgtgg tgggcacaat caaggatctg aagcagggct
4681 atctgagcca ggtcatccac gagatcgtgg acctgatgat ccactaccag gccgtggtgg
4741 tgctggagaa cctgaatttc ggctttaaga gcaagaggac cggcatcgcc gagaaggccg
4801 tgtaccagca gttcgagaag atgctgatcg ataagctgaa ttgcctggtg ctgaaggact
4861 atccagcaga gaaagtggga ggcgtgctga acccatacca gctgacagac cagttcacct
4921 cctttgccaa gatgggcacc cagtctggct tcctgtttta cgtgcctgcc ccatatacat
4981 ctaagatcga tcccctgacc ggcttcgtgg accccttcgt gtggaaaacc atcaagaatc
5041 acgagagccg caagcacttc ctggagggct tcgactttct gcactacgac gtgaaaaccg
5101 gcgacttcat cctgcacttt aagatgaaca gaaatctgtc cttccagagg ggcctgcccg
5161 gctttatgcc tgcatgggat atcgtgttcg agaagaacga gacacagttt gacgccaagg
5221 gcaccccttt catcgccggc aagagaatcg tgccagtgat cgagaatcac agattcaccg
5281 gcagataccg ggacctgtat cctgccaacg agctgatcgc cctgctggag gagaagggca
5341 tcgtgttcag ggatggctcc aacatcctgc caaagctgct ggagaatgac gattctcacg
5401 ccatcgacac catggtggcc ctgatccgca gcgtgctgca gatgcggaac tccaatgccg
5461 ccacaggcga ggactatatc aacagccccg tgcgcgatct gaatggcgtg tgcttcgact
5521 cccggtttca gaacccagag tggcccatgg acgccgatgc caatggcgcc taccacatcg
5581 ccctgaaggg ccagctgctg ctgaatcacc tgaaggagag caaggatctg aagctgcaga
5641 acggcatctc caatcaggac tggctggcct acatccagga gctgcgcaac aaaaggccgg
5701 cggccacgaa aaaggccggc caggcaaaaa agaaaaaggg atcctaccca tacgatgttc
5761 cagattacgc ttatccctac gacgtgcctg attatgcata cccatatgat gtccccgact
5821 atgcctaaag cggccgcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg
5881 tggccaatgc cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg
5941 gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca
6001 ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa
6061 atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat gcccatatgc
6121 tggctgccat gaacaaaggt tggctataaa gaggtcatca gtatatgaaa cagccccctg
6181 ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt
6241 gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca
6301 gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc ttatggagat
6361 ccctcgacct gcagcccaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
6421 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
6481 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
6541 tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc atagtcccgc
6601 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
6661 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
6721 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagcta acttgtttat
6781 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
6841 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
6901 gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
6961 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
7021 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
7081 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
7141 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
7201 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
7261 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
7321 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
7381 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
7441 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
7501 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
7561 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
7621 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
7681 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
7741 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
7801 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
7861 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
7921 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
7981 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
8041 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
8101 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
8161 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
8221 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
8281 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
8341 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
8401 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
8461 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
8521 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
8581 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
8641 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
8701 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
8761 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
8821 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
8881 aaagtgccac ctg
//
