| 质粒名称: | pCMV-MMLVgag-3xNES-Cas9 |
|---|---|
| 目录编号: | 181752 |
| 存储实验室: | David Liu |
|
载体骨架基本信息 |
|
| 载体名称: | pCDNA3-like |
| 载体出品公司: | Clontech |
| 空载体大小: | 4757bp |
| 完整质粒大小: | 10783 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | Ampicillin, 100 μg/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: | MMLV-Gag-3xNES-Cas9 (Spy) |
| 物种来源: | H. sapiens (human), M. musculus (mouse) |
| 插入大小: | 4757 bp |
| 启动子: | CMV |
| 标签/融合蛋白: | Flag (N terminal on insert) |
| 克隆方法: | Ligation Independent Cloning |
| 5’测序引物 | ACCCTGGTCATCATCCTG |
| 3’测序引物 | CAGGAAACAGCTATGAC; CCTTCCGAGTGAGAGAC |
|
文献引用方法 |
|
| 材料和方法部分: | pCMV-MMLVgag-3xNES-Cas9 was a gift from David Liu (Addgene plasmid # 181752 ; http://n2t.net/addgene:181752 ; RRID:Addgene_181752) |
| 参考文献部分: | Engineered virus-like particles for efficient in vivo delivery of therapeutic proteins. Banskota S, Raguram A, Suh S, Du SW, Davis JR, Choi EH, Wang X, Nielsen SC, Newby GA, Randolph PB, Osborn MJ, Musunuru K, Palczewski K, Liu DR. Cell. 2022 Jan 7. pii: S0092-8674(21)01484-7. doi: 10.1016/j.cell.2021.12.021. 10 |
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 pCMV-MMLVgag-3xNES-Cas9 10783 bp ds-DNA circular SYN 08-FEB-2022
DEFINITION MMLV-Gag fused to 3x NES-SpyCas9 with the TSTLLMENSS cleavage site
in between NES and Cas9. For production of virus like particles with
Cas9 nuclease RNP cargo..
ACCESSION .
VERSION .
KEYWORDS pCMV-MMLVgag-3xNES-Cas9
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10783)
AUTHORS Banskota S, Raguram A, Suh S, Du SW, Davis JR, Choi EH, Wang X,
Nielsen SC, Newby GA, Randolph PB, Osborn MJ, Musunuru K, Palczewski
K, Liu DR
TITLE Engineered virus-like particles for efficient in vivo delivery of
therapeutic proteins.
JOURNAL Cell. 2022 Jan 7. pii: S0092-8674(21)01484-7. doi:
10.1016/j.cell.2021.12.021.
PUBMED 35021064
REFERENCE 2 (bases 1 to 10783)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Feb 8, 2022 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..10783
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 4..420
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 350..372
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 388..404
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
CDS 1633..1659
/codon_start=1
/product="nuclear export signal from the HIV Rev protein
(Fischer et al., 1995)"
/label=NES
/translation="LPPLERLTL"
CDS 1672..1698
/codon_start=1
/product="nuclear export signal from the HIV Rev protein
(Fischer et al., 1995)"
/label=NES
/translation="LPPLERLTL"
CDS 1711..1737
/codon_start=1
/product="nuclear export signal from the HIV Rev protein
(Fischer et al., 1995)"
/label=NES
/translation="LPPLERLTL"
CDS 1771..1794
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 1828..1848
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1861..5961
/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 6004..6024
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
primer_bind complement(6030..6049)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 6095..6150
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(6149..6168)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(6508..6524)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6508..6524)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6521..6543)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6532..6548
/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(6556..6586)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6601..6622
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6739..6756)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6910..7498)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6990..7009)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7669..8529)
/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 8292..8311
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8530..8634)
/gene="bla"
/label=AmpR promoter
rep_origin complement(8660..9115)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(8797..8818)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 9009..9028
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 9242..9264
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 9256..9273
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 9257..9273
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 9283..9302
/label=T7
/note="T7 promoter, forward primer"
promoter 9283..9301
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
enhancer 9422..9801
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 9802..10005
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 9956..9976
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 10002..10026
/label=LNCX
/note="Human CMV promoter, forward primer"
intron 10117..10689
/label=-beta-globin intron
/note="intron from rabbit beta-globin gene"
ORIGIN
1 atgggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa agatgtcgag
61 cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac cttctgctct
121 gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa ccgagacctc
181 atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc agaccaggtc
241 ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt caagcccttt
301 gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc ccttgaacct
361 cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc tctaggcgcc
421 aaacctaaac ctcaagttct ttctgacagt ggggggccgc tcatcgacct acttacagaa
481 gaccccccgc cttataggga cccaagacca cccccttccg acagggacgg aaatggtgga
541 gaagcgaccc ctgcgggaga ggcaccggac ccctccccaa tggcatctcg cctacgtggg
601 agacgggagc cccctgtggc cgactccact acctcgcagg cattccccct ccgcgcagga
661 ggaaacggac agcttcaata ctggccgttc tcctcttctg acctttacaa ctggaaaaat
721 aataaccctt ctttttctga agatccaggt aaactgacag ctctgatcga gtctgtcctc
781 atcacccatc agcccacctg ggacgactgt cagcagctgt tggggactct gctgaccgga
841 gaagaaaaac aacgggtgct cttagaggct agaaaggcgg tgcggggcga tgatgggcgc
901 cccactcaac tgcccaatga agtcgatgcc gcttttcccc tcgagcgccc agactgggat
961 tacaccaccc aggcaggtag gaaccaccta gtccactatc gccagttgct cctagcgggt
1021 ctccaaaacg cgggcagaag ccccaccaat ttggccaagg taaaaggaat aacacaaggg
1081 cccaatgagt ctccctcggc cttcctagag agacttaagg aagcctatcg caggtacact
1141 ccttatgacc ctgaggaccc agggcaagaa actaatgtgt ctatgtcttt catttggcag
1201 tctgccccag acattgggag aaagttagag aggttagaag atttaaaaaa caagacgctt
1261 ggagatttgg ttagagaggc agaaaagatc tttaataaac gagaaacccc ggaagaaaga
1321 gaggaacgta tcaggagaga aacagaggaa aaagaagaac gccgtaggac agaggatgag
1381 cagaaagaga aagaaagaga tcgtaggaga catagagaga tgagcaagct attggccact
1441 gtcgttagtg gacagaaaca ggatagacag ggaggagaac gaaggaggtc ccaactcgat
1501 cgcgaccagt gtgcctactg caaagaaaag gggcactggg ctaaagattg tcccaagaaa
1561 ccacgaggac ctcggggacc aagaccccag acctccctcc tgaccctaga tgactctggc
1621 ggctcacttc aactgcctcc acttgaaaga ctgacactgg gatcattaca attacctcct
1681 ttagaacgat taacactcgg ttcactacag cttccgcctc ttgagagatt gacattaacg
1741 tccacgctgc taatggagaa ctcgtctgga gattacaaag acgatgacga taagaaacgg
1801 acagccgacg gaagcgagtt cgagtcacca aagaagaagc ggaaagtcag cggggggtca
1861 gacaagaagt acagcatcgg cctggacatc ggcaccaact ctgtgggctg ggccgtgatc
1921 accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac
1981 agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc
2041 acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat
2101 ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg
2161 gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac
2221 atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa
2281 ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg
2341 atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg
2401 gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc
2461 aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg
2521 ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg
2581 attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat
2641 gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag
2701 atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg
2761 ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg
2821 atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag
2881 cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc
2941 tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa
3001 aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag
3061 cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc
3121 attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag
3181 aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga
3241 ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg
3301 gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac
3361 ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat
3421 aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc
3481 ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg
3541 aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc
3601 ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc
3661 aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg
3721 accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac
3781 ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg
3841 ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat
3901 ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc
3961 ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac
4021 gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg
4081 aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc
4141 gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg
4201 aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg
4261 gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat
4321 atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc
4381 gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac
4441 aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac
4501 tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc
4561 aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg
4621 gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact
4681 aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag
4741 ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac
4801 caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac
4861 cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg
4921 atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac
4981 atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct
5041 ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc
5101 accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag
5161 acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc
5221 agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat
5281 tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa
5341 gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt
5401 ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac
5461 tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag
5521 aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac
5581 tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag
5641 cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc
5701 ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc
5761 atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct
5821 gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag
5881 gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac
5941 ctgtctcagc tgggaggtga ctctggcggc tcaaaaagaa ccgccgacgg cagcgaattc
6001 gagcccaaga agaagaggaa agtctgagat ctttttccct ctgccaaaaa ttatggggac
6061 atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca
6121 atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat
6181 ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct
6241 gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc
6301 cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt
6361 gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact agccagattt
6421 ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg gagatccctc
6481 gacggatcgg ccgcaattcg taaccatgtc atagctgttt cctgtgtgaa attgttatcc
6541 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
6601 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
6661 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
6721 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
6781 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
6841 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
6901 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
6961 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
7021 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
7081 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
7141 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
7201 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
7261 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
7321 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
7381 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
7441 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
7501 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
7561 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
7621 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
7681 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
7741 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
7801 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
7861 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
7921 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
7981 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
8041 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
8101 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
8161 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
8221 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
8281 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
8341 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
8401 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
8461 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
8521 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
8581 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
8641 tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
8701 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
8761 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
8821 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
8881 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
8941 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
9001 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
9061 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat
9121 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
9181 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
9241 tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga ctcactatag
9301 ggcgaattgg agctccaccg cggtggcggc cgctctagag agcttggccc attgcatacg
9361 ttgtatccat atcataatat gtacatttat attggctcat gtccaacatt accgccatgt
9421 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
9481 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
9541 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
9601 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
9661 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
9721 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
9781 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
9841 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
9901 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
9961 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt
10021 cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgggaccga
10081 tccagcctcc ggtcgaccga tcctgagaac ttcagggtga gtttggggac ccttgattgt
10141 tctttctttt tcgctattgt aaaattcatg ttatatggag ggggcaaagt tttcagggtg
10201 ttgtttagaa tgggaagatg tcccttgtat caccatggac cctcatgata attttgtttc
10261 tttcactttc tactctgttg acaaccattg tctcctctta ttttcttttc attttctgta
10321 actttttcgt taaactttag cttgcatttg taacgaattt ttaaattcac ttttgtttat
10381 ttgtcagatt gtaagtactt tctctaatca cttttttttc aaggcaatca gggtatatta
10441 tattgtactt cagcacagtt ttagagaaca attgttataa ttaaatgata aggtagaata
10501 tttctgcata taaattctgg ctggcgtgga aatattctta ttggtagaaa caactacacc
10561 ctggtcatca tcctgccttt ctctttatgg ttacaatgat atacactgtt tgagatgagg
10621 ataaaatact ctgagtccaa accgggcccc tctgctaacc atgttcatgc cttcttctct
10681 ttcctacagc tcctgggcaa cgtgctggtt gttgtgctgt ctcatcattt tggcaaagaa
10741 ttcctcgagg gccggatccg aattcctgta tttgtctgaa aac
//
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