| 质粒名称: | pCMV-Hs.cargo(Cas9) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 174864 |
| 存储实验室: | Feng Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1 |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | 10649 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | SpCas9 flanked by HsPeg10 UTRs |
| 物种来源: | H. sapiens (human); S.pyogenes |
| 插入大小: | 5200 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | CMV |
| 标签/融合蛋白: |
HA-SV40 NLS（插入件上的N端子）
SV40 NLS（插入件上的C端子）
|
| 克隆方法: | Gibson克隆 |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | CMV Forward |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pCMV-Hs.cargo(Cas9) was a gift from Feng Zhang (Addgene plasmid # 174864 ; http://n2t.net/addgene:174864 ; RRID:Addgene_174864) |
| 参考文献部分: | Mammalian retrovirus-like protein PEG10 packages its own mRNA and can be pseudotyped for mRNA delivery. Segel M, Lash B, Song J, Ladha A, Liu CC, Jin X, Mekhedov SL, Macrae RK, Koonin EV, Zhang F. Science. 2021 Aug 20;373(6557):882-889. doi: 10.1126/science.abg6155. 10.1126/science.abg6155 PubMed 34413232 |
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
载体图谱.png)
载体简介
载体序列
LOCUS pCMV-Hs.cargo(Cas9) 10634 bp ds-DNA circular SYN 14-SEP-2021
DEFINITION Expresses human Peg10 cargoRNA encoding SpCas9.
ACCESSION .
VERSION .
KEYWORDS pCMV-Hs.cargo(Cas9)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10634)
AUTHORS Segel M, Lash B, Song J, Ladha A, Liu CC, Jin X, Mekhedov SL, Macrae
RK, Koonin EV, Zhang F
TITLE Mammalian retrovirus-like protein PEG10 packages its own mRNA and
can be pseudotyped for mRNA delivery.
JOURNAL Science. 2021 Aug 20;373(6557):882-889. doi:
10.1126/science.abg6155.
PUBMED 34413232
REFERENCE 2 (bases 1 to 10634)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 14, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..10634
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 629..648
/label=T7
/note="T7 promoter, forward primer"
promoter 629..647
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1165..1191
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 1195..1215
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1225..5325
/codon_start=1
/product="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
/label=Cas9
/note="generates RNA-guided double strand breaks in DNA"
/translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 5329..5349
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
primer_bind complement(5991..6008)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5997..6221
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 6267..6695
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6354..6373)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6564..6585
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(6704..6724)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 6709..7038
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 6889..7024
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 6951..6970
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 7105..7899
/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(7159..7178)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 7769..7788
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 8073..8194
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(8110..8129)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 8164..8183
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(8243..8259)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8243..8259)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(8256..8278)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 8267..8283
/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(8291..8321)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8336..8357
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(8474..8491)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(8645..9233)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(8725..8744)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(9404..10264)
/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 10027..10046
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(10265..10369)
/gene="bla"
/label=AmpR promoter
primer_bind complement(10444..10463)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
661 gctagcgttt aaaaccctcc tcggtgcaac ctatataagg ctcacagtct gcgctcctgg
721 tacacgcgct tcaacttcgg ttggtgtgtg tcgaagaaac ctgactgcgc cctgaggaga
781 acagcggaga aggtccaccg agcctggcga aaggtccgct gagcgggctg tcgtccggag
841 ccactccggg ctgcggagca cccagtggag accgcgcctg gctcaggtgt gggaccccat
901 ccttcctgtc ttcgcagagg agtcctcgcg tgaaataagc gggttttgaa aacaaaaaaa
961 agaaggagtg gaagaggggg ccaggatcca ggcctccatc cccacagaag tgaagctaca
1021 gctgggaggt ctcctcccac cccaaccgtc accctgggtc ccgactgccc acctcctcct
1081 cctccccctc cccccaacaa caacaacaac aacaactcca agcacaccgg ccataagagt
1141 gcgtgtgtcc ccaacgccac catgtaccca tacgatgttc cagattacgc ttcgccgaag
1201 aaaaagcgca aggtcgaagc gtccgacaag aagtacagca tcggcctgga catcggcacc
1261 aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag
1321 gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc
1381 gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc
1441 agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg
1501 gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac
1561 gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc
1621 accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg
1681 atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac
1741 ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac
1801 cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct
1861 gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag
1921 aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag
1981 agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac
2041 gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc
2101 aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc
2161 aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc
2221 ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac
2281 cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga agagttctac
2341 aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg
2401 aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag
2461 atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg
2521 aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc
2581 cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc
2641 accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag
2701 cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg
2761 ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga
2821 atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc
2881 aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag
2941 tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca
3001 taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag
3061 gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag
3121 gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg
3181 cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag
3241 cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc
3301 atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg
3361 tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt
3421 aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg
3481 cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga
3541 cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc
3601 cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg
3661 tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg
3721 tccgactacg atgtggacca tatcgtgcct cagagctttc tgaaggacga ctccatcgac
3781 aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa
3841 gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc
3901 cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag
3961 gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag
4021 atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg
4081 aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac
4141 aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg
4201 ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac
4261 aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc
4321 gccaagtact tcttctacag caacatcatg aactttttca agaccgagat taccctggcc
4381 aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg
4441 tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat
4501 atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag
4561 aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc
4621 ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag
4681 tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc
4741 ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac
4801 ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg
4861 ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg
4921 aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag
4981 cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc
5041 agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc
5101 tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt
5161 accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg
5221 aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc
5281 ggcctgtacg agacacggat cgacctgtct cagctgggag gcgacagccc caagaagaag
5341 agaaaggtgg aggccagcta aatacctgtc atgtccttca ggatctctgc cctcaaaatt
5401 tattcctgtt cagcttctca atcagtgact gtgtgctaaa ttttaggcta ctgtatcttc
5461 aggccacctg aggcacatcc tctctgaaac ggctatggaa ggttagggcc actctggact
5521 ggcacacatc ctaaagcacc aaaagacctt caacattttc tgagagcaac agagtatttg
5581 ccaataaatg atctctcatt tttccacctt gactgccaat ctaactaaaa taattaataa
5641 gtttactttc cagccagtcc tggaagtctg ggttttacct gccaaaacct ccatcaccat
5701 ctaaattata ggctgccaaa tttgctgttt aacatttaca gagaagctga tacaaacgca
5761 ggaaatgctg atttctttat ggagggggag acgaggagga ggaggacatg acttttcttg
5821 cggtttcggt accctctttt taaatcactg gaggactgag gccttattaa ggaagcctta
5881 agcttggtac cgagctcgga tccactagtc cagtgtggtg gaattctgca gatatccagc
5941 acagtggcgg ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt
6001 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
6061 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
6121 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
6181 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
6241 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
6301 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
6361 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
6421 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
6481 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
6541 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
6601 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
6661 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
6721 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
6781 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
6841 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
6901 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
6961 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
7021 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
7081 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
7141 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
7201 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
7261 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
7321 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
7381 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
7441 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
7501 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
7561 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
7621 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
7681 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
7741 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
7801 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
7861 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
7921 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
7981 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
8041 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
8101 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
8161 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
8221 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
8281 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
8341 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
8401 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
8461 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
8521 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
8581 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
8641 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
8701 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
8761 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
8821 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
8881 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
8941 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
9001 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
9061 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
9121 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
9181 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
9241 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
9301 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
9361 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
9421 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
9481 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
9541 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
9601 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
9661 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
9721 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
9781 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
9841 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
9901 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
9961 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
10021 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
10081 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
10141 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
10201 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
10261 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
10321 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
10381 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
10441 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
10501 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
10561 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
10621 cagatatacg cgtt
//
