| 质粒名称: | pBA904 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 122238 |
| 存储实验室: | Jonathan Weissman |
|
载体骨架基本信息 |
|
| 载体名称: | pU6-sgRNA EF1Alpha-puro-T2A-BFP (Addgene, #60955) |
| 完整质粒大小: | 8914 bp |
| 载体修饰: | 使用BlpI和XhoI位点插入修饰的sgRNA（在恒定区/tracr的茎环2中具有捕获序列1）。 |
| 载体类型: | 哺乳细胞表达,慢病毒,CRISPR |
| 筛选标记: | 嘌呤霉素,BFP |
| 载体抗性: | 氨苄霉素100 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | sgRNA with cs1 in stem loop 2 of the constant region |
| 物种来源: | Synthetic |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BlpI (未破坏) |
| 3’克隆位点: | XhoI(未破坏) |
| 5’测序引物 | cagcacaaaaggaaactcacc |
|
文献引用方法 |
|
| 材料和方法部分: | pBA904 was a gift from Jonathan Weissman (Addgene plasmid # 122238 ; http://n2t.net/addgene:122238 ; RRID:Addgene_122238) |
| 参考文献部分: | Combinatorial single-cell CRISPR screens by direct guide RNA capture and targeted sequencing. Replogle JM, Norman TM, Xu A, Hussmann JA, Chen J, Cogan JZ, Meer EJ, Terry JM, Riordan DP, Srinivas N, Fiddes IT, Arthur JG, Alvarado LJ, Pfeiffer KA, Mikkelsen TS, Weissman JS, Adamson B. Nat Biotechnol. 2020 Aug;38(8):954-961. doi: 10.1038/s41587-020-0470-y. Epub 2020 Mar 30. 10.1038/s41587-020-0470-y PubMed 32231336 |
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 pBA904 8903 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Lentiviral CRISPR guide vector expressing a eGFP-NT2 sgRNA with cs1
incorporated in the loop of the sgRNA constant region..
ACCESSION .
VERSION .
KEYWORDS pBA904
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8903)
TITLE Direct capture of CRISPR guides enables scalable, multiplexed, and
multi-omic Perturb-seq
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 8903)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8903
/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 3142..3353
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
intron 3354..4292
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 4249..4269
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
CDS 4313..4909
/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(4313..4332)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 4809..4829
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
CDS 4919..4972
/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 4985..5683
/codon_start=1
/product="monomeric blue fluorescent protein"
/label=TagBFP
/note="mammalian codon-optimized"
/translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
LPSKLGHKLN"
protein_bind 5703..5736
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
misc_feature 5792..6380
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(5845..5865)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(6263..6274)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(6383..6399)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6384..6400)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 6909..7089
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
rep_origin complement(7151..7739)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7231..7250)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7910..8770)
/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 8533..8552
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8771..8875)
/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 aacttggccg ctttaaggcc ggtcctagca aggccaagtg gcaccgagtc
3061 ggtgcttttt ttctcgagta ctaggatcca ttaggcggcc gcgtggataa ccgtattacc
3121 gccatgcatg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
3181 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
3241 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
3301 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
3361 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
3421 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
3481 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
3541 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
3601 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
3661 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
3721 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
3781 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
3841 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
3901 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
3961 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
4021 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
4081 attagttctc gcgcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
4141 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
4201 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
4261 cagtggttca aagttttttt cttccatttc aggtgtcgtg agctagccca ccatgaccga
4321 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct
4381 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat
4441 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
4501 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
4561 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
4621 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
4681 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
4741 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
4801 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
4861 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgccg gatcgggaga
4921 gggcagagga agtctgctaa catgcggtga cgtcgaggag aatcctggcc caccggtcgc
4981 caccagcgag ctgattaagg agaacatgca catgaagctg tacatggagg gcaccgtgga
5041 caaccatcac ttcaagtgca catccgaggg cgaaggcaag ccctacgagg gcacccagac
5101 catgagaatc aaggtggtcg agggcggccc tctccccttc gccttcgaca tcctggctac
5161 tagcttcctc tacggcagca agaccttcat caaccacacc cagggcatcc ccgacttctt
5221 caagcagtcc ttccctgagg gcttcacatg ggagagagtc accacatacg aagacggggg
5281 cgtgctgacc gctacccagg acaccagcct ccaggacggc tgcctcatct acaacgtcaa
5341 gatcagaggg gtgaacttca catccaacgg ccctgtgatg cagaagaaaa cactcggctg
5401 ggaggccttc accgagacgc tgtaccccgc tgacggcggc ctggaaggca gaaacgacat
5461 ggccctgaag ctcgtgggcg ggagccatct gatcgcaaac atcaagacca catatagatc
5521 caagaaaccc gctaagaacc tcaagatgcc tggcgtctac tatgtggact acagactgga
5581 aagaatcaag gaggccaaca acgagaccta cgtcgagcag cacgaggtgg cagtggccag
5641 atactgcgac ctccctagca aactggggca caagcttaat taagaattcg tcgagggacc
5701 taataacttc gtatagcata cattatacga agttatacat gtttaagggt tccggttcca
5761 ctaggtacaa ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga
5821 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
5881 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
5941 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
6001 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
6061 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
6121 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
6181 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
6241 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
6301 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
6361 cctttgggcc gcctccccgc atcgataccg tcgacctcga tcgagaccta gaaaaacatg
6421 gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg ctagaagcac
6481 aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga ccaatgactt
6541 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
6601 ttcactccca acgaagacaa gatatccttg atctgtggat ctaccacaca caaggctact
6661 tccctgattg gcagaactac acaccagggc cagggatcag atatccactg acctttggat
6721 ggtgctacaa gctagtacca gttgagcaag agaaggtaga agaagccaat gaaggagaga
6781 acacccgctt gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtat
6841 tagagtggag gtttgacagc cgcctagcat ttcatcacat ggcccgagag ctgcatccgg
6901 actgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
6961 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
7021 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
7081 tctctagcag catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
7141 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
7201 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
7261 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
7321 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
7381 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
7441 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
7501 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
7561 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
7621 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
7681 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
7741 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
7801 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
7861 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
7921 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
7981 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
8041 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
8101 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
8161 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
8221 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
8281 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
8341 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
8401 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
8461 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
8521 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
8581 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
8641 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
8701 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
8761 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
8821 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
8881 ttccccgaaa agtgccacct gac
//
