| 质粒名称: | pU6-sgRNA EF1Alpha-puro-T2A-BFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 60955 |
| 存储实验室: | Jonathan Weissman |
|
载体骨架基本信息 |
|
| 载体名称: | pSICO衍生物 |
| 载体出品公司: | Addgene |
| 空载体大小: | 8200 bp |
| 完整质粒大小: | 8888 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，慢病毒，CRISPR |
| 筛选标记: | -- |
| 载体抗性: | 氨苄青霉素，100μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | Stbl3 |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | sgGFP-NT2 |
| 物种来源: | -- |
| 插入大小: | 113 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pU6-sgRNA EF1Alpha-puro-T2A-BFP was a gift from Jonathan Weissman (Addgene plasmid # 60955 ; http://n2t.net/addgene:60955 ; RRID:Addgene_60955) |
| 参考文献部分: | Genome-Scale CRISPR-Mediated Control of Gene Repression and Activation. Gilbert LA, Horlbeck MA, Adamson B, Villalta JE, Chen Y, Whitehead EH, Guimaraes C, Panning B, Ploegh HL, Bassik MC, Qi LS, Kampmann M, Weissman JS. Cell. 2014 Oct 23;159(3):647-61. doi: 10.1016/j.cell.2014.09.029. Epub 2014 Oct 9. 10.1016/j.cell.2014.09.029 PubMed 25307932 |
买家导航
pU6-sgRNA EF1Alpha-puro-T2A-BFP载体质粒基本信息
pU6-sgRNA EF1Alpha-puro-T2A-BFP质粒图谱载体图谱和pU6-sgRNA EF1Alpha-puro-T2A-BFP载体序列质粒序列多克隆位点信息
pU6-sgRNA EF1Alpha-puro-T2A-BFP质粒载体简介
pU6-sgRNA EF1Alpha-puro-T2A-BFP质粒序列载体序列
LOCUS pU6-sgRNA EF1Alpha-puro-T2A-BFP 8877 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Parental vector for the CRISPRi/a libraries. Expresses an sgRNA from
the U6 promoter and a puromycin resistance cassette and BFP from the
EF1Alpha promoter.
ACCESSION .
VERSION .
KEYWORDS pU6-sgRNA EF1Alpha-puro-T2A-BFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8877)
AUTHORS Gilbert LA, Horlbeck MA, Adamson B, Villalta JE, Chen Y, Whitehead
EH, Guimaraes C, Panning B, Ploegh HL, Bassik MC, Qi LS, Kampmann M,
Weissman JS
TITLE Genome-Scale CRISPR-Mediated Control of Gene Repression and
Activation.
JOURNAL Cell. 2014 Oct 23;159(3):647-61. doi: 10.1016/j.cell.2014.09.029.
Epub 2014 Oct 9.
PUBMED 25307932
REFERENCE 2 (bases 1 to 8877)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8877
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(47..66)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 238..617
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 619..817
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 772..792
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 835..1015
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1062..1187
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1684..1917
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 2102..2146
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 2444..2561
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
protein_bind 2814..2847
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
primer_bind 2851..2871
/label=mU6-F
/note="Mouse U6 promoter, forward primer"
promoter 3116..3327
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
intron 3328..4266
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 4223..4243
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
CDS 4287..4883
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(4287..4306)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 4783..4803
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
CDS 4893..4946
/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"
CDS 4959..5657
/codon_start=1
/product="monomeric blue fluorescent protein"
/label=TagBFP
/note="mammalian codon-optimized"
/translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
LPSKLGHKLN"
protein_bind 5677..5710
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
misc_feature 5766..6354
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(5819..5839)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(6237..6248)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(6357..6373)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6358..6374)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 6883..7063
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
rep_origin complement(7125..7713)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7205..7224)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7884..8744)
/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 8507..8526
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8745..8849)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
1501 aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga
1561 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
1621 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
1681 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
1741 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
1801 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
1861 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
1921 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
1981 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
2041 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
2101 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
2161 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
2221 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
2281 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
2341 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact
2401 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg
2461 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
2521 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
2581 gcagagatcc agtttggtta gtaccgggcc cgctctagag atccgacgcg ccatctctag
2641 gcccgcgccg gccccctcgc acggacttgt gggagaagct cggctactcc cctgccccgg
2701 ttaatttgca tataatattt cctagtaact atagaggctt aatgtgcgat aaaagacaga
2761 taatctgttc tttttaatac tagctacatt ttacatgata ggcttggatt tctataactt
2821 cgtatagcat acattatacg aagttataaa cagcacaaaa ggaaactcac cctaactgta
2881 aagtaattgt gtgttttgag actataagta tcccttggag aaccaccttg ttggaccagg
2941 atgggcacca cccgtttaag agctaagctg gaaacagcat agcaagttta aataaggcta
3001 gtccgttatc aacttgaaaa agtggcaccg agtcggtgct ttttttctcg agtactagga
3061 tccattaggc ggccgcgtgg ataaccgtat taccgccatg catgtgcccg tcagtgggca
3121 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt
3181 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt
3241 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt
3301 cgcaacgggt ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc
3361 ctctttacgg gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg
3421 attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa
3481 ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg
3541 cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa
3601 aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc
3661 caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg
3721 tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg
3781 gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc
3841 ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg
3901 cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg
3961 ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac
4021 tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgcgctt ttggagtacg
4081 tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg
4141 gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt
4201 gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca
4261 tttcaggtgt cgtgagctag cccaccatga ccgagtacaa gcccacggtg cgcctcgcca
4321 cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
4381 ccacgcgcca caccgtcgac ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
4441 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
4501 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
4561 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc
4621 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc
4681 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg
4741 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
4801 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
4861 gcatgacccg caagcccggt gccggatcgg gagagggcag aggaagtctg ctaacatgcg
4921 gtgacgtcga ggagaatcct ggcccaccgg tcgccaccag cgagctgatt aaggagaaca
4981 tgcacatgaa gctgtacatg gagggcaccg tggacaacca tcacttcaag tgcacatccg
5041 agggcgaagg caagccctac gagggcaccc agaccatgag aatcaaggtg gtcgagggcg
5101 gccctctccc cttcgccttc gacatcctgg ctactagctt cctctacggc agcaagacct
5161 tcatcaacca cacccagggc atccccgact tcttcaagca gtccttccct gagggcttca
5221 catgggagag agtcaccaca tacgaagacg ggggcgtgct gaccgctacc caggacacca
5281 gcctccagga cggctgcctc atctacaacg tcaagatcag aggggtgaac ttcacatcca
5341 acggccctgt gatgcagaag aaaacactcg gctgggaggc cttcaccgag acgctgtacc
5401 ccgctgacgg cggcctggaa ggcagaaacg acatggccct gaagctcgtg ggcgggagcc
5461 atctgatcgc aaacatcaag accacatata gatccaagaa acccgctaag aacctcaaga
5521 tgcctggcgt ctactatgtg gactacagac tggaaagaat caaggaggcc aacaacgaga
5581 cctacgtcga gcagcacgag gtggcagtgg ccagatactg cgacctccct agcaaactgg
5641 ggcacaagct taattaagaa ttcgtcgagg gacctaataa cttcgtatag catacattat
5701 acgaagttat acatgtttaa gggttccggt tccactaggt acaattcgat atcaagctta
5761 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
5821 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
5881 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
5941 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
6001 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
6061 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
6121 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
6181 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
6241 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
6301 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
6361 accgtcgacc tcgatcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca
6421 gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca
6481 gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
6541 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatatc
6601 cttgatctgt ggatctacca cacacaaggc tacttccctg attggcagaa ctacacacca
6661 gggccaggga tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag
6721 caagagaagg tagaagaagc caatgaagga gagaacaccc gcttgttaca ccctgtgagc
6781 ctgcatggga tggatgaccc ggagagagaa gtattagagt ggaggtttga cagccgccta
6841 gcatttcatc acatggcccg agagctgcat ccggactgta ctgggtctct ctggttagac
6901 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
6961 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
7021 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagcatgtg agcaaaaggc
7081 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
7141 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
7201 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
7261 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
7321 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
7381 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
7441 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
7501 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
7561 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
7621 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
7681 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
7741 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
7801 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
7861 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
7921 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
7981 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
8041 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
8101 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
8161 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
8221 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
8281 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
8341 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
8401 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
8461 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
8521 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
8581 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
8641 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
8701 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
8761 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
8821 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgac
//
