| 质粒名称: | pgRNA-CKB,pmU6-gRNA-CAG-mKate2-T2A-Bsd |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 73501 |
| 存储实验室: | Bruce Conklin |
|
载体骨架基本信息 |
|
| 载体名称: | pSico |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 9596 bp |
| 载体类型: | 哺乳动物表达，慢病毒，CRISPR |
| 筛选标记: | 杀稻瘟菌素 |
| 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: | mKate2-T2A-Bsd |
| 物种来源: | H. sapiens (human), Synthetic |
| 启动子 | CAG |
| 标签/融合蛋白: | NLS（插入件上的N端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | EcoRI（未销毁） |
| 3' 克隆位点: | EcoRI（未销毁） |
| 5' 测序引物 | CCTCTGCTAACCATGTTCATGC |
| 3' 测序引物 | GATCTACCACATTTGTAGAG |
|
文献引用方法 |
|
| 材料和方法部分: | pgRNA-CKB was a gift from Bruce Conklin (Addgene plasmid # 73501 ; http://n2t.net/addgene:73501 ; RRID:Addgene_73501) |
| 参考文献部分: | CRISPR Interference Efficiently Induces Specific and Reversible Gene Silencing in Human iPSCs. Mandegar MA, Huebsch N, Frolov EB, Shin E, Truong A, Olvera MP, Chan AH, Miyaoka Y, Holmes K, Spencer CI, Judge LM, Gordon DE, Eskildsen TV, Villalta JE, Horlbeck MA, Gilbert LA, Krogan NJ, Sheikh SP, Weissman JS, Qi LS, So PL, Conklin BR. Cell Stem Cell. 2016 Apr 7;18(4):541-53. doi: 10.1016/j.stem.2016.01.022. Epub 2016 Mar 10. 10.1016/j.stem.2016.01.022 PubMed 26971820 |
买家导航
pgRNACKB载体基本信息
pgRNACKB载体图谱质粒图谱和多克隆位点信息
pgRNACKB载体简介
pgRNACKB载体序列
LOCUS pgRNA-CKB 9596 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION gRNA expression vector (with mKate2).
ACCESSION .
VERSION .
KEYWORDS pgRNA-CKB
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9596)
AUTHORS Mandegar MA, Huebsch N, Frolov EB, Shin E, Truong A, Olvera MP, Chan
AH, Miyaoka Y, Holmes K, Spencer CI, Judge LM, Gordon DE, Eskildsen
TV, Villalta JE, Horlbeck MA, Gilbert LA, Krogan NJ, Sheikh SP,
Weissman JS, Qi LS, So PL, Conklin BR
TITLE CRISPR Interference Efficiently Induces Specific and Reversible Gene
Silencing in Human iPSCs.
JOURNAL Cell Stem Cell. 2016 Apr 7;18(4):541-53. doi:
10.1016/j.stem.2016.01.022. Epub 2016 Mar 10.
PUBMED 26971820
REFERENCE 2 (bases 1 to 9596)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9596
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 309..426
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
protein_bind 679..712
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
primer_bind 716..736
/label=mU6-F
/note="Mouse U6 promoter, forward primer"
enhancer 1053..1356
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 1358..1635
/label=chicken beta-actin promoter
intron 1637..2654
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 2578..2600
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 2662..2681
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
CDS 2719..2739
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 2743..2763
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 2767..2787
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 2803..3498
/codon_start=1
/product="monomeric far-red fluorescent protein (Shcherbo
et al., 2009)"
/label=mKate2
/note="mammalian codon-optimized"
/translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
CDLPSKLGHR"
CDS 3499..3552
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
primer_bind complement(3996..4015)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
protein_bind 4250..4283
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
misc_feature 4339..4927
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4392..4412)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4810..4821)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(4930..4946)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4931..4947)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 5456..5636
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
rep_origin complement(5698..6286)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5778..5797)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6457..7317)
/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 7080..7099
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7318..7422)
/gene="bla"
/label=AmpR promoter
primer_bind complement(7506..7525)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 7697..8076
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 8078..8276
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 8231..8251
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 8294..8474
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 8521..8646
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 9145..9378
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
ORIGIN
1 gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata aaattattca
61 taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga
121 atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg
181 gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca
241 ttcgattagt gaacggatcg gcactgcgtg cgccaattct gcagacaaat ggcagtattc
301 atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
361 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
421 aattttcggg tttattacag ggacagcaga gatccagttt ggttagtacc gggcccgctc
481 tagagatccg acgcgccatc tctaggcccg cgccggcccc ctcgcacgga cttgtgggag
541 aagctcggct actcccctgc cccggttaat ttgcatataa tatttcctag taactataga
601 ggcttaatgt gcgataaaag acagataatc tgttcttttt aatactagct acattttaca
661 tgataggctt ggatttctat aacttcgtat agcatacatt atacgaagtt ataaacagca
721 caaaaggaaa ctcaccctaa ctgtaaagta attgtgtgtt ttgagactat aagtatccct
781 tggagaacca ccttgttggg gagacggacg tctccgttta agagctatgc tggaaacagc
841 atagcaagtt taaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
901 ctttttttct cgagtactag gatccattag gcggccgcgt ggataaccgt attaccgcca
961 tgcatcccgg gtgactgact gagcggccgc actagttatt aatagtaatc aattacgggg
1021 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
1081 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
1141 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
1201 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
1261 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
1321 cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc cacgttctgc
1381 ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa
1441 ttattttgtg cagcgatggg ggcggggggg gggggggggc gcgcgccagg cggggcgggg
1501 cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc
1561 gcgctccgaa agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga
1621 agcgcgcggc gggcggggag tcgctgcgac gctgccttcg ccccgtgccc cgctccgccg
1681 ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc
1741 gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt
1801 ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc ggggggagcg
1861 gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc
1921 ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg
1981 aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag
2041 gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc
2101 tgcaaccccc cctgcacccc cctccccgag ttgctgagca cggcccggct tcgggtgcgg
2161 ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg
2221 gggtgccggg cggggcgggg ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg
2281 gcccccggag cgccggcggc tgtcgaggcg cggcgagccg cagccattgc cttttatggt
2341 aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg
2401 ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc gccggcagga
2461 aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc
2521 agcctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt
2581 tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt catgccttct
2641 tctttttcct acagctcctg ggcaacgtgc tggttattgt gctgtctcat cattttggca
2701 aagaattccg ccaccatgcc aaaaaagaag agaaaggtag atccaaaaaa gaagagaaag
2761 gtagatccaa aaaagaagag aaaggtacac gtgtcaacgc gtatggtgag cgagctgatt
2821 aaggagaaca tgcacatgaa gctgtacatg gagggcaccg tgaacaacca ccacttcaag
2881 tgcacatccg agggcgaagg caagccctac gagggcaccc agaccatgag aatcaaggcg
2941 gtcgagggcg gccctctccc cttcgccttc gacatcctgg ctaccagctt catgtacggc
3001 agcaaaacct tcatcaacca cacccagggc atccccgact tctttaagca gtccttcccc
3061 gagggcttca catgggagag agtcaccaca tacgaagacg ggggcgtgct gaccgctacc
3121 caggacacca gcctccagga cggctgcctc atctacaacg tcaagatcag aggggtgaac
3181 ttcccatcca acggccctgt gatgcagaag aaaacactcg gctgggaggc ctccaccgag
3241 accctgtacc ccgctgacgg cggcctggaa ggcagagccg acatggccct gaagctcgtg
3301 ggcgggggcc acctgatctg caacttgaag accacataca gatccaagaa acccgctaag
3361 aacctcaaga tgcccggcgt ctactatgtg gacagaagac tggaaagaat caaggaggcc
3421 gacaaagaga cctacgtcga gcagcacgag gtggctgtgg ccagatactg cgacctccct
3481 agcaaactgg ggcacagaga gggcagagga agtcttctaa catgcggtga cgtggaggag
3541 aatcccggcc ctatgaagac cttcaacatc tcccagcagg atctagaatt agtagaagta
3601 gcgacagaga agattacaat gctttatgag gataataaac atcatgtggg agcggcaatt
3661 cgtacgaaaa caggagaaat catttcggca gtacatattg aagcgtatat aggacgagta
3721 actgtttgtg cagaagccat tgcgattggt agtgcagttt cgaatggaca aaaggatttt
3781 gacacgattg tagctgttag acacccttat tctgacgaag tagatagaag tattcgagtg
3841 gtaagtcctt gtggtatgtg tagggagttg atttcagact atgcaccaga ttgttttgtg
3901 ttaatagaaa tgaatggcaa gttagtcaaa actacgattg aagaactcat tccactcaaa
3961 tatacccgaa attaatcgac atgataagat acattgatga gtttggacaa accacaacta
4021 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtga
4081 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
4141 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
4201 caagtaaaac ctctacaaat gtggtagatc gaattcgtcg agggacctaa taacttcgta
4261 tagcatacat tatacgaagt tatacatgtt taagggttcc ggttccacta ggtacaattc
4321 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
4381 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
4441 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
4501 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
4561 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
4621 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
4681 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
4741 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
4801 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
4861 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
4921 tccccgcatc gataccgtcg acctcgatcg agacctagaa aaacatggag caatcacaag
4981 tagcaataca gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga
5041 ggtgggtttt ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt
5101 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
5161 aagacaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattggca
5221 gaactacaca ccagggccag ggatcagata tccactgacc tttggatggt gctacaagct
5281 agtaccagtt gagcaagaga aggtagaaga agccaatgaa ggagagaaca cccgcttgtt
5341 acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtattag agtggaggtt
5401 tgacagccgc ctagcatttc atcacatggc ccgagagctg catccggact gtactgggtc
5461 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
5521 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
5581 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagcat
5641 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
5701 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
5761 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
5821 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
5881 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
5941 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
6001 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
6061 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
6121 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
6181 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
6241 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
6301 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
6361 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
6421 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
6481 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
6541 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
6601 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
6661 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
6721 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
6781 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
6841 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
6901 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
6961 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
7021 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
7081 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
7141 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
7201 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
7261 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
7321 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
7381 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
7441 gccacctgac gcttaagcgg tcgacggatc gggagatctc ccgatcccct atggtgcact
7501 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
7561 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
7621 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
7681 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
7741 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
7801 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
7861 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
7921 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
7981 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
8041 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
8101 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
8161 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
8221 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg
8281 ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
8341 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
8401 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
8461 gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag
8521 ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga
8581 ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag
8641 agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca
8701 gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga
8761 ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag
8821 ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca
8881 accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag
8941 atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgg ccgcgctgat
9001 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
9061 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
9121 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
9181 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
9241 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
9301 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
9361 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
9421 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
9481 ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
9541 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaat
//
