| 质粒名称: | pCMV-T7-SpG-HF1-P2A-EGFP (RTW5000) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 139996 |
| 存储实验室: | Benjamin Kleinstiver |
|
载体骨架基本信息 |
|
| 载体名称: | RTW3027 |
| 载体出品公司: | Kleinstiver Lab (爱迪基因 ID 139987) |
| 载体抗性: | 氨苄青霉素 |
| 载体类型: | 哺乳动物表达，CRISPR；体外转录；T7启动子 |
| 筛选标记: | -- |
| 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: | human codon optimized SpCas9 variant named SpG-HF1 with BPNLS-3xFLAG-P2A-EGFP,RTW5000 |
| 物种来源: | Synthetic |
| 基因突变: | SpG=D1135L/S1136W/G1218K/E1219Q/R1335Q/T1337R; HF1=N497A/R661A/Q695A/Q926A |
| 启动子 | CMV and T7 |
| 标签/融合蛋白: | BPNLS-3XLAGG-P2A-EGFP（插入件上的C端子） |
| 克隆方法: | Gibson 克隆 |
| 5' 测序引物 | oBK6928-CCAAGTCTCCACCCCATTGACG |
| 3' 测序引物 | oBK219-GGGAGTGGCACCTTCCAGGGTC |
|
文献引用方法 |
|
| 材料和方法部分: | pCMV-T7-SpG-HF1-P2A-EGFP (RTW5000) was a gift from Benjamin Kleinstiver (Addgene plasmid # 139996 ; http://n2t.net/addgene:139996 ; RRID:Addgene_139996) |
| 参考文献部分: | Unconstrained genome targeting with near-PAMless engineered CRISPR-Cas9 variants. Walton , Russell T., Christie, Kathleen A., Whittaker, Madelynn N., Kleinstiver, Benjamin P.. Science , 26 Mar 2020 10.1126/science.aba8853 |
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
质粒图谱.png)
载体简介
载体序列
LOCUS pCMV-T7-SpG-HF1-P2A-EGFP (RTW5000) 8451 bp ds-DNA circular SYN 20-MAR-2020
DEFINITION CMV and T7 promoter expression plasmid for human codon optimized
SpG-HF1(SpG=D1135L/S1136W/G1218K/E1219Q/R1335Q/T1337R;HF1=N497A/R661
A/Q695A/Q926A) with a c-terminal bi-partite NLS, 3x flag tag, and
P2A-EGFP.
ACCESSION .
VERSION .
KEYWORDS pCMV-T7-SpG-HF1-P2A-EGFP (RTW5000)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8451)
AUTHORS Kleinstiver
TITLE Kleinstiver lab CRISPR plasmids
JOURNAL unpublished
REFERENCE 2 (bases 1 to 8451)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Mar 20, 2020 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8451
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 4036..4056
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 4069..4134
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 4153..4209
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS 4210..4929
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(4255..4276)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(4516..4535)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 4863..4884
/label=EGFP-C
/note="EGFP, forward primer"
CDS 4935..4952
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(4975..4992)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 4981..5205
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(5276..5292)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5276..5292)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5289..5311)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5300..5316
/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(5324..5354)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5369..5390
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5507..5524)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5678..6266)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5758..5777)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6437..7297)
/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 7060..7079
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7298..7402)
/gene="bla"
/label=AmpR promoter
primer_bind complement(7481..7500)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 7672..8051
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 8052..8255
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 8206..8226
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 8297..8316
/label=T7
/note="T7 promoter, forward primer"
promoter 8297..8315
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 8323..8332
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 8329..1596
/codon_start=1
/product="N-terminal portion of Streptococcus pyogenes Cas9
(Zetsche et al., 2015)"
/label=Cas9(N)
/translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTAFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
ORIGIN
1 agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc
61 acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat
121 ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg
181 gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac
241 atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa
301 ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg
361 atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg
421 gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc
481 aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg
541 ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg
601 attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat
661 gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag
721 atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg
781 ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg
841 atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag
901 cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc
961 tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa
1021 aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag
1081 cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc
1141 attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag
1201 aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga
1261 ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg
1321 gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccgcctt cgataagaac
1381 ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat
1441 aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc
1501 ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg
1561 aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc
1621 ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc
1681 aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg
1741 accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac
1801 ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcgcc
1861 ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat
1921 ttcctgaagt ccgacggctt cgccaacaga aacttcatgg ccctgatcca cgacgacagc
1981 ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac
2041 gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg
2101 aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc
2161 gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg
2221 aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg
2281 gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat
2341 atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc
2401 gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac
2461 aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac
2521 tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc
2581 aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg
2641 gtggaaaccc gggccatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact
2701 aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag
2761 ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac
2821 caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac
2881 cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg
2941 atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac
3001 atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct
3061 ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc
3121 accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag
3181 acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc
3241 agaaagaagg actgggaccc taagaagtac ggcggcttcc tgtggcccac cgtggcctat
3301 tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa
3361 gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt
3421 ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac
3481 tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccaa gcagctgcag
3541 aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac
3601 tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag
3661 cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc
3721 ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc
3781 atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct
3841 gccgccttca agtactttga caccaccatc gaccggaagc agtacagaag caccaaagag
3901 gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac
3961 ctgtctcagc tgggaggcga cggatccggc ggaggcggaa gcgggaaaag aaccgccgac
4021 ggcagcgaat tcgagcccaa gaagaagagg aaagtctcga gcggaggcga ctacaaagac
4081 catgacggtg attataaaga tcatgacatc gattacaagg atgacgatga caagggaggc
4141 ggtggaagcg gcgcaacaaa cttctctctg ctgaaacaag ccggagatgt cgaagagaat
4201 cctggaccga tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
4261 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
4321 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
4381 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
4441 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
4501 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
4561 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
4621 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
4681 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
4741 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
4801 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
4861 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
4921 tacaagtgac cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga
4981 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
5041 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
5101 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
5161 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
5221 gaaccagctg gggctcgata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
5281 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
5341 gcataaagtg taaagcctag ggtgcctaat gagtgagcta actcacatta attgcgttgc
5401 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
5461 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
5521 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
5581 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
5641 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
5701 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
5761 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
5821 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
5881 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
5941 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
6001 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
6061 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
6121 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
6181 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
6241 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
6301 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
6361 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
6421 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
6481 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
6541 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
6601 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
6661 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
6721 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
6781 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
6841 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
6901 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
6961 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
7021 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
7081 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
7141 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
7201 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
7261 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
7321 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
7381 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
7441 cgggagatcg atctcccgat cccctagggt cgactctcag tacaatctgc tctgatgccg
7501 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
7561 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
7621 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat
7681 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
7741 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
7801 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
7861 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
7921 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
7981 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
8041 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
8101 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
8161 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
8221 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
8281 agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat ggacaagaag
8341 tacagcatcg gcctggacat cggcaccaac tctgtgggct gggccgtgat caccgacgag
8401 tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca c
//
