| 质粒名称: | pTE4254 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 89050 |
| 存储实验室: | Ervin Welker |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.3-TOPO |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5805 bp |
| 完整质粒大小: | 9202 bp |
| 载体类型: | 哺乳细胞表达,CRISPR |
| 筛选标记: | 新霉素（G418） |
| 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: | St chimeric gRNA,Streptococcus thermophilus guideRNA |
| 插入大小: | 152 bp |
| 启动子 | human U6 |
| 5' 测序引物 | AAAACAGGAAGGCAAAATGC |
| 3' 测序引物 | ggtcgggcaggaagaggg |
|
文献引用方法 |
|
| 材料和方法部分: | pTE4254 was a gift from Ervin Welker (Addgene plasmid # 89050 ; http://n2t.net/addgene:89050 ; RRID:Addgene_89050) |
| 参考文献部分: | Cpf1 nucleases demonstrate robust activity to induce DNA modification by exploiting homology directed repair pathways in mammalian cells. Toth E, Weinhardt N, Bencsura P, Huszar K, Kulcsar PI, Talas A, Fodor E, Welker E. Biol Direct. 2016 Sep 14;11:46. doi: 10.1186/s13062-016-0147-0. 10.1186/s13062-016-0147-0 [pii] PubMed 27630115 |
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 pTE4254 9204 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expresses St chimeric gRNA and human codon-optimized StCas9.
ACCESSION .
VERSION .
KEYWORDS pTE4254
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9204)
AUTHORS Toth E, Weinhardt N, Bencsura P, Huszar K, Kulcsar PI, Talas A,
Fodor E, Welker E
TITLE Cpf1 nucleases demonstrate robust activity to induce DNA
modification by exploiting homology directed repair pathways in
mammalian cells.
JOURNAL Biol Direct. 2016 Sep 14;11:46. doi: 10.1186/s13062-016-0147-0.
PUBMED 27630115
REFERENCE 2 (bases 1 to 9204)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9204
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 18..397
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 398..601
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 552..572
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 598..622
/label=LNCX
/note="Human CMV promoter, forward primer"
CDS 716..4078
/codon_start=1
/product="Cas9 endonuclease from the Streptococcus
thermophilus Type II CRISPR/Cas system (Esvelt et al.,
2013)"
/label=St1Cas9
/note="generates RNA-guided double strand breaks in DNA"
/translation="MSDLVLGLDIGIGSVGVGILNKVTGEIIHKNSRIFPAAQAENNLV
RRTNRQGRRLARRKKHRRVRLNRLFEESGLITDFTKISINLNPYQLRVKGLTDELSNEE
LFIALKNMVKHRGISYLDDASDDGNSSVGDYAQIVKENSKQLETKTPGQIQLERYQTYG
QLRGDFTVEKDGKKHRLINVFPTSAYRSEALRILQTQQEFNPQITDEFINRYLEILTGK
RKYYHGPGNEKSRTDYGRYRTSGETLDNIFGILIGKCTFYPDEFRAAKASYTAQEFNLL
NDLNNLTVPTETKKLSKEQKNQIINYVKNEKAMGPAKLFKYIAKLLSCDVADIKGYRID
KSGKAEIHTFEAYRKMKTLETLDIEQMDRETLDKLAYVLTLNTEREGIQEALEHEFADG
SFSQKQVDELVQFRKANSSIFGKGWHNFSVKLMMELIPELYETSEEQMTILTRLGKQKT
TSSSNKTKYIDEKLLTEEIYNPVVAKSVRQAIKIVNAAIKEYGDFDNIVIEMARETNED
DEKKAIQKIQKANKDEKDAAMLKAANQYNGKAELPHSVFHGHKQLATKIRLWHQQGERC
LYTGKTISIHDLINNSNQFEVDHILPLSITFDDSLANKVLVYATANQEKGQRTPYQALD
SMDDAWSFRELKAFVRESKTLSNKKKEYLLTEEDISKFDVRKKFIERNLVDTLYASRVV
LNALQEHFRAHKIDTKVSVVRGQFTSQLRRHWGIEKTRDTYHHHAVDALIIAASSQLNL
WKKQKNTLVSYSEDQLLDIETGELISDDEYKESVFKAPYQHFVDTLKSKEFEDSILFSY
QVDSKFNRKISDATIYATRQAKVGKDKADETYVLGKIKDIYTQDGYDAFMKIYKKDKSK
FLMYRHDPQTFEKVIEPILENYPNKQINDKGKEVPCNPFLKYKEEHGYIRKYSKKGNGP
EIKSLKYYDSKLGNHIDITPKDSNNKVVLQSVSPWRADVYFNKTTGKYEILGLKYADLQ
FDKGTGTYKISQEKYNDIKKKEGVDSDSEFKFTLYKNDLLLVKDTETKEQQLFRFLSRT
MPKQKHYVELKPYDKQKFEGGEALIKVLGNVANSGQCKKGLGKSNISIYKVRTDVLGNQ
HIIKNEGDKPKLDF"
CDS 4091..4111
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
primer_bind complement(4167..4186)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4211..4259
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4461..4889
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4548..4567)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4758..4779
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4898..4918)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 4903..5232
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 5083..5218
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5145..5164
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 5299..6093
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(5353..5372)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5963..5982
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 6269..6390
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6306..6325)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 6360..6379
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(6439..6455)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6439..6455)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6452..6474)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6463..6479
/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(6487..6517)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6532..6553
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6670..6687)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6841..7429)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6921..6940)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7600..8460)
/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 8223..8242
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8461..8565)
/gene="bla"
/label=AmpR promoter
primer_bind complement(8640..8659)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
misc_RNA complement(8776..8900)
/label=St1 gRNA scaffold
/note="guide RNA scaffold for the Streptococcus
thermophilus CRISPR/Cas9 system (Esvelt et al., 2013)"
promoter complement(8929..9169)
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind complement(8979..8998)
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
primer_bind complement(9149..9169)
/label=hU6-F
/note="Human U6 promoter, forward primer"
ORIGIN
1 atgctagttg gcgcgttgac attgattatt gactagttat taatagtaat caattacggg
61 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
121 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
181 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
241 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
301 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
361 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
421 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
481 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
541 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
601 tcgtttagtg aaccgtcaga tcgcctggga tccccatcca cgctgttttg acctccatag
661 aagacaccgg gaccgatcca gcctccggac tctagaggat cgaacccttg ccaccatgag
721 cgacctggtg ctgggcctgg acatcggcat cggcagcgtg ggcgtgggca tcctgaacaa
781 ggtgaccggc gagatcatcc acaagaacag tcgcatcttc cctgctgctc aggctgagaa
841 caacctggtg cgccgcacca accgccaggg tcgccggctt gctcgccgca agaagcaccg
901 gcgcgtgcgc ctgaaccgcc tgttcgagga gagcggcctg atcaccgact tcaccaagat
961 cagcatcaac ctgaacccct accagctgcg cgtgaagggc ctgaccgacg agctgagcaa
1021 cgaggagctg ttcatcgccc tgaagaacat ggtgaagcac cgcggcatca gctacctgga
1081 cgacgccagc gacgacggca acagcagcgt gggcgactac gcccagatcg tgaaggagaa
1141 cagcaagcag ctggagacca agacccccgg ccagatccag ctggagcgct accagaccta
1201 cggccagctg cgcggcgact tcaccgtgga gaaggacggc aagaagcacc gcctgatcaa
1261 cgtgttcccc accagcgcct accgcagcga ggccctgcgc atcctgcaga cccagcagga
1321 gttcaacccc cagatcaccg acgagttcat caaccgctac ctggagatcc tgaccggcaa
1381 gcgcaagtac taccacggcc ccggcaacga gaagagccgc accgactacg gccgctaccg
1441 caccagcggc gagaccctgg acaacatctt cggcatcctg atcggcaagt gcaccttcta
1501 ccccgacgag ttccgcgccg ccaaggccag ctacaccgcc caggagttca acctgctgaa
1561 cgacctgaac aacctgaccg tgcccaccga gaccaagaag ctgagcaagg agcagaagaa
1621 ccagatcatc aactacgtga agaacgagaa ggccatgggc cccgccaagc tgttcaagta
1681 catcgccaag ctgctgagct gcgacgtggc cgacatcaag ggctaccgca tcgacaagag
1741 cggcaaggcc gagatccaca ccttcgaggc ctaccgcaag atgaagaccc tggagaccct
1801 ggacatcgag cagatggacc gagagaccct ggacaagctg gcctacgtgc tgaccctgaa
1861 caccgagcgc gagggcatcc aggaggccct ggagcacgag ttcgccgacg gcagcttcag
1921 ccagaagcag gtggacgagc tggtgcagtt ccgcaaggcc aacagcagca tcttcggcaa
1981 gggctggcac aacttcagcg tgaagctgat gatggagctg atccccgagc tgtacgagac
2041 cagcgaggag cagatgacca tcctgacccg cctgggcaag cagaagacca ccagcagcag
2101 caacaagacc aagtacatcg acgagaagct gctgaccgag gagatctaca accccgtggt
2161 ggccaagagc gtgcgccagg ccatcaagat cgtgaacgcc gccatcaagg agtacggcga
2221 cttcgacaac atcgtgatcg agatggcccg cgagaccaac gaggacgacg agaagaaggc
2281 catccagaag atccagaagg ccaacaagga cgagaaggac gccgccatgc tgaaggccgc
2341 caaccagtac aacggcaagg ccgagctgcc ccacagcgtg ttccacggcc acaagcagct
2401 ggccaccaag atccgcctgt ggcaccagca gggcgagcgc tgcctgtaca ccggcaagac
2461 catcagcatc cacgacctga tcaacaacag caaccagttc gaggtggacc acatcctgcc
2521 cctgagcatc accttcgacg acagcctggc caacaaggtg ctggtgtacg ccaccgccaa
2581 ccaggagaag ggccagcgca ccccctacca ggccctggac agcatggacg acgcctggag
2641 cttccgcgag ctgaaggcct tcgtgcgcga gagcaagacc ctgagcaaca agaagaagga
2701 gtacctgctg accgaggagg acatcagcaa gttcgacgtg cgcaagaagt tcatcgagcg
2761 caacctggtg gacaccctct acgccagccg cgtggtgctg aacgccctgc aggagcactt
2821 ccgcgcccac aagatcgaca ccaaggtgag cgtggtgcgc ggccagttca ccagccagct
2881 gcgccgccac tggggcatcg agaagacccg cgacacctac caccaccacg ccgtggacgc
2941 cctgatcatt gcggcttcta gccagctgaa cctgtggaag aagcagaaga acaccctggt
3001 gagctacagc gaggaccagc tgctggacat cgagaccggc gagctgatca gcgacgacga
3061 gtacaaggag agcgtgttca aggcccccta ccagcacttc gtggacaccc tgaagagcaa
3121 ggagttcgag gacagcatcc tgttcagcta ccaggtggac agcaagttca accgcaagat
3181 cagcgacgcc accatctacg ccacccgcca ggccaaggtg ggcaaggaca aggccgacga
3241 gacctacgtg ctgggcaaga tcaaggacat ctacacccag gacggctacg acgccttcat
3301 gaagatctac aagaaggaca agagcaagtt cctgatgtac cgccacgacc cccagacctt
3361 cgagaaggtg atcgagccca tcctggagaa ctaccccaac aagcagatca acgataaagg
3421 caaggaggtg ccctgcaacc ccttcctgaa gtacaaggag gagcacggct acatccgcaa
3481 gtacagcaag aagggcaacg gccccgagat caagagcctg aagtactacg acagcaagct
3541 gggcaaccac atcgacatca cccccaagga cagcaacaac aaggtggtgc tgcagagcgt
3601 gagcccctgg cgcgccgacg tgtacttcaa caagaccacc ggcaagtacg agatcctggg
3661 cctgaagtac gccgacctgc agtttgataa gggcaccggc acctacaaga tcagccagga
3721 gaagtacaac gacatcaaga agaaggaggg cgtggacagc gacagcgagt tcaagttcac
3781 cctgtacaag aacgaccttc tgctggtgaa ggacaccgag accaaggagc aacagctgtt
3841 ccgcttcctg agccgcacca tgcccaagca gaagcactac gtggagctga agccctacga
3901 caagcagaag ttcgagggcg gcgaggccct gatcaaggtg ctgggcaacg tggccaacag
3961 cggccagtgc aagaagggcc tgggcaagag caacatcagc atctacaagg tgcgcaccga
4021 cgtgctgggc aaccagcaca tcatcaagaa cgagggcgac aagcccaagt tggacttcag
4081 cagggctgac cccaagaaga agaggaaggt gtgagcaggg ttcgatccct accggttagt
4141 aatgagttta aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
4201 cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca
4261 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
4321 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
4381 cccagggctc gcagccaacg tcggggcggc agaccctgcc atagcagatc tgcgcagctg
4441 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
4501 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
4561 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
4621 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
4681 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
4741 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
4801 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
4861 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
4921 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
4981 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
5041 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
5101 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
5161 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
5221 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
5281 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
5341 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
5401 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
5461 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
5521 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
5581 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
5641 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
5701 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
5761 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
5821 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
5881 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
5941 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
6001 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
6061 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac
6121 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
6181 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
6241 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
6301 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
6361 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
6421 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
6481 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
6541 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
6601 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
6661 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
6721 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
6781 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
6841 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
6901 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
6961 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
7021 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
7081 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
7141 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
7201 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
7261 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
7321 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
7381 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
7441 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
7501 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
7561 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
7621 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
7681 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
7741 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
7801 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
7861 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
7921 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
7981 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
8041 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
8101 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
8161 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
8221 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
8281 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
8341 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
8401 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
8461 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
8521 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
8581 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
8641 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
8701 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
8761 tgcttaagaa aaaaacaccc tgccataaaa tgacagggtg ttgatttcgg catgaagcct
8821 tatctttgta gcttctgcaa gatttaagta actgtgtaag tttcgttgta cagttactta
8881 aatcttgaga gtacaaaaac ggagacgggt acccgtctcc ggtgtttcgt cctttccaca
8941 agatatataa agccaagaaa tcgaaatact ttcaagttac ggtaagcata tgatagtcca
9001 ttttaaaaca taattttaaa actgcaaact acccaagaaa ttattacttt ctacgtcacg
9061 tattttgtac taatatcttt gtgtttacag tcaaattaat tctaattatc tctctaacag
9121 ccttgtatcg tatatgcaaa tatgaaggaa tcatgggaaa taggccctct tcctgcccga
9181 ccttagatcc gaatccatat cgat
//
