| 质粒名称: | pCMV-T7-ABEmax(7.10)-SpCas9-NG-P2A-EGFP (RTW4564) |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 140005 |
| 存储实验室: | Benjamin Kleinstiver |
|
载体骨架基本信息 |
|
| 载体名称: | pCMV_ABEmax_P2A_GFP |
| 载体出品公司: | David Liu (Addgene Plasmid 112101) |
| 载体类型: | Mammalian Expression, CRISPR ; in vitro transcription; T7 |
| 载体抗性: | 氨苄霉素 |
| 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: | human codon optimized ABEmax(7.10) SpCas9-NG with P2A-EGFP/RTW4564 |
| 物种来源: | Synthetic |
| 基因突变: | nSpCas9=D10A; SpCas9-NG=L1111R/D1135V/G1218R/E1219F/A1322R/R1335V/T1337R |
| 启动子: | CMV and T7 |
| 标签/融合蛋白: | P2A-EGFP (C terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5’测序引物 | roBK6928-CCAAGTCTCCACCCCATTGACG |
| 3’测序引物 | BK219-GGGAGTGGCACCTTCCAGGGTC |
|
文献引用方法 |
|
| 材料和方法部分: | pCMV-T7-ABEmax(7.10)-SpCas9-NG-P2A-EGFP (RTW4564) was a gift from Benjamin Kleinstiver (Addgene plasmid # 140005 ; http://n2t.net/addgene:140005 ; RRID:Addgene_140005) |
| 参考文献部分: | Unconstrained genome targeting with near-PAMless engineered CRISPR-Cas9 variants. Walton RT, Christie KA, Whittaker MN, Kleinstiver BP. Science. 2020 Apr 17;368(6488):290-296. doi: 10.1126/science.aba8853. Epub 2020 Mar 26. 10.1126/science.aba8853 PubMed 32217751 |
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
-SpCas9-NG-P2A-EGFP (RTW4564)140005-质粒图谱.png)
载体简介
载体序列
LOCUS pCMV-T7-ABEmax(7.10)-SpCas9-NG-P2A-EGFP (RTW4564) 9627 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION CMV and T7 promoter expression plasmid for human codon optimized
ABEmax(7.10) A-to-G base editor with
SpCas9-NG(D10A/L1111R/D1135V/G1218R/E1219F/A1322R/R1335V/T1337R) and
P2A-EGFP.
ACCESSION .
VERSION .
KEYWORDS pCMV-T7-ABEmax(7.10)-SpCas9-NG-P2A-EGFP (RTW4564)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9627)
AUTHORS Walton RT, Christie KA, Whittaker MN, Kleinstiver BP
TITLE Unconstrained genome targeting with near-PAMless engineered
CRISPR-Cas9 variants.
JOURNAL Science. 2020 Apr 17;368(6488):290-296. doi:
10.1126/science.aba8853. Epub 2020 Mar 26.
PUBMED 32217751
REFERENCE 2 (bases 1 to 9627)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..9627
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 521..540
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(759..863)
/gene="bla"
/label=AmpR promoter
primer_bind complement(942..961)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 1133..1512
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 1513..1716
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 1667..1687
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 1758..1777
/label=T7
/note="T7 promoter, forward primer"
promoter 1758..1776
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1826..1846
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 3035..4750
/codon_start=1
/product="N-terminal portion of Streptococcus pyogenes Cas9
(Zetsche et al., 2015)"
/label=Cas9(N)
/translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
CDS 7178..7198
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7208..7264
/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 7265..7981
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(7310..7331)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(7571..7590)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 7918..7939
/label=EGFP-C
/note="EGFP, forward primer"
CDS 7994..8014
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 8023..8040
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(8063..8080)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 8069..8293
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(8364..8380)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8364..8380)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(8377..8399)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 8388..8404
/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(8412..8442)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8457..8478
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(8595..8612)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(8766..9354)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(8846..8865)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(9525..758)
/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"
ORIGIN
1 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
61 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
121 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
181 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
241 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
301 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
361 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
421 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
481 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
541 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
601 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
661 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
721 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
781 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
841 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
901 tcgggagatc gatctcccga tcccctaggg tcgactctca gtacaatctg ctctgatgcc
961 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
1021 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
1081 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
1141 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
1201 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
1261 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
1321 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
1381 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
1441 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
1501 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
1561 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
1621 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
1681 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc
1741 tagagatccg cggccgctaa tacgactcac tatagggaga gccgccacca tgaaacggac
1801 agccgacgga agcgagttcg agtcaccaaa gaagaagcgg aaagtctctg aagtcgagtt
1861 tagccacgag tattggatga ggcacgcact gaccctggca aagcgagcat gggatgaaag
1921 agaagtcccc gtgggcgccg tgctggtgca caacaataga gtgatcggag agggatggaa
1981 caggccaatc ggccgccacg accctaccgc acacgcagag atcatggcac tgaggcaggg
2041 aggcctggtc atgcagaatt accgcctgat cgatgccacc ctgtatgtga cactggagcc
2101 atgcgtgatg tgcgcaggag caatgatcca cagcaggatc ggaagagtgg tgttcggagc
2161 acgggacgcc aagaccggcg cagcaggctc cctgatggat gtgctgcacc accccggcat
2221 gaaccaccgg gtggagatca cagagggaat cctggcagac gagtgcgccg ccctgctgag
2281 cgatttcttt agaatgcgga gacaggagat caaggcccag aagaaggcac agagctccac
2341 cgactctgga ggatctagcg gaggatcctc tggaagcgag acaccaggca caagcgagtc
2401 cgccacacca gagagctccg gcggctcctc cggaggatcc tctgaggtgg agttttccca
2461 cgagtactgg atgagacatg ccctgaccct ggccaagagg gcacgcgatg agagggaggt
2521 gcctgtggga gccgtgctgg tgctgaacaa tagagtgatc ggcgagggct ggaacagagc
2581 catcggcctg cacgacccaa cagcccatgc cgaaattatg gccctgagac agggcggcct
2641 ggtcatgcag aactacagac tgattgacgc caccctgtac gtgacattcg agccttgcgt
2701 gatgtgcgcc ggcgccatga tccactctag gatcggccgc gtggtgtttg gcgtgaggaa
2761 cgcaaaaacc ggcgccgcag gctccctgat ggacgtgctg cactaccccg gcatgaatca
2821 ccgcgtcgaa attaccgagg gaatcctggc agatgaatgt gccgccctgc tgtgctattt
2881 ctttcggatg cctagacagg tgttcaatgc tcagaagaag gcccagagct ccaccgactc
2941 cggaggatct agcggaggct cctctggctc tgagacacct ggcacaagcg agagcgcaac
3001 acctgaaagc agcgggggca gcagcggggg gtcagacaag aagtacagca tcggcctggc
3061 catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa
3121 gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc
3181 cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag
3241 aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat
3301 ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga
3361 taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga
3421 gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga
3481 cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat
3541 cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca
3601 gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc
3661 catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc
3721 cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc
3781 caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac
3841 ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt
3901 tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac
3961 cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca
4021 ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat
4081 tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga
4141 agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct
4201 cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat
4261 cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta
4321 cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta
4381 ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga
4441 ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag
4501 cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa
4561 gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt
4621 gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga
4681 cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa
4741 gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc
4801 cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga
4861 ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga
4921 gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca
4981 gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat
5041 ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa
5101 cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa
5161 agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag
5221 ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt
5281 gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac
5341 ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga
5401 gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa
5461 gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat
5521 caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc tgaaggacga
5581 ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt
5641 gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa
5701 gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga
5761 actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca
5821 cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat
5881 ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt
5941 ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa
6001 cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta
6061 cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg
6121 caaggctacc gccaagtact tcttctacag caacatcatg aactttttca agaccgagat
6181 taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg
6241 ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc
6301 ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat
6361 cagacccaag aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa
6421 gtacggcggc ttcgtgagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga
6481 aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga
6541 aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt
6601 gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg
6661 gaagagaatg ctggcctctg ccagattcct gcagaaggga aacgaactgg ccctgccctc
6721 caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga
6781 ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat
6841 cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt
6901 gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat
6961 ccacctgttt accctgacca atctgggagc ccctagagcc ttcaagtact ttgacaccac
7021 catcgaccgg aaggtgtaca gaagcaccaa agaggtgctg gacgccaccc tgatccacca
7081 gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag gtgactctgg
7141 cggctcaaaa agaaccgccg acggcagcga attcgagccc aagaagaaga ggaaagtcgg
7201 aagcggagct actaacttca gcctgctgaa gcaggctgga gacgtggagg agaaccctgg
7261 acctatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
7321 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
7381 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
7441 caccctcgtg accaccctga cctatggagt gcagtgcttc agccgctacc ccgaccacat
7501 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
7561 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
7621 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
7681 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
7741 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
7801 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
7861 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
7921 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
7981 gtctggtggt tctcccaaga agaagaggaa agtctaaccg gtcatcatca ccatcaccat
8041 tgagtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
8101 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
8161 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
8221 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
8281 gtgggctcta tggcttctga ggcggaaaga accagctggg gctcgatacc gtcgacctct
8341 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
8401 acaattccac acaacatacg agccggaagc ataaagtgta aagcctaggg tgcctaatga
8461 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
8521 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
8581 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
8641 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
8701 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
8761 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
8821 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
8881 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
8941 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
9001 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
9061 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
9121 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
9181 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
9241 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
9301 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
9361 cctttgatct tttctacggg gtctgacact cagtggaacg aaaactcacg ttaagggatt
9421 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
9481 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
9541 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
9601 gtcgtgtaga taactacgat acgggag
//
