| 质粒名称: | pCAG-CBE4max-SpRY-P2A-EGFP (RTW5133) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 139999 |
| 存储实验室: | Benjamin Kleinstiver |
|
载体骨架基本信息 |
|
| 载体名称: | pRZ33 |
| 载体出品公司: | Ketih Joung |
| 载体抗性: | 氨苄青霉素 |
| 载体类型: | 哺乳细胞表达,CRISPR |
| 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 CBE4max SpCas9 variant named SpRY with P2A-EGFP，RTW5133 |
| 物种来源: | Synthetic |
| 基因突变: | nSpCas9=D10A; SpRY=A61R/L1111R/D1135L/S1136W/G1218K/E1219Q/N1317R/A1322R/R1333P/R1335Q/T1337R |
| 启动子 | CAG |
| 标签/融合蛋白: | P2A-EGFP（插入件上的C端子） |
| 克隆方法: | Gibson克隆 |
| 5' 测序引物 | oBK280-CCTACAGCTCCTGGGCAACG |
| 3' 测序引物 | oBK23-CAGAGGGAAAAAGATCTCAGTGG |
|
文献引用方法 |
|
| 材料和方法部分: | pCAG-CBE4max-SpRY-P2A-EGFP (RTW5133) was a gift from Benjamin Kleinstiver (Addgene plasmid # 139999 ; http://n2t.net/addgene:139999 ; RRID:Addgene_139999) |
| 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 pCAG-CBE4max-SpRY-P2A-EGFP (RTW5133) 11195 bp ds-DNA circular SYN 20-MAR-2020
DEFINITION CAG promoter expression plasmid for human codon optimized BE4max
C-to-T base editor with
SpRY(D10A/A61R/L1111R/D1135L/S1136W/G1218K/E1219Q/N1317R/A1322R/R133
3P/R1335Q/T1337R) and P2A-EGFP.
ACCESSION .
VERSION .
KEYWORDS pCAG-CBE4max-SpRY-P2A-EGFP (RTW5133)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11195)
AUTHORS Kleinstiver
TITLE Kleinstiver lab CRISPR plasmids
JOURNAL unpublished
REFERENCE 2 (bases 1 to 11195)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Mar 20, 2020 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..11195
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 37..416
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 418..693
/label=chicken beta-actin promoter
intron 694..1710
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1634..1656
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1718..1737
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
CDS 1845..1865
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1866..2549
/codon_start=1
/product="cytidine deaminase (C to U editing enzyme) from
rat"
/label=APOBEC-1
/note="can use ssDNA as a substrate (Komor et al., 2016)"
/translation="SSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGG
RHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLS
RYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNE
AHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHIL
WATGLK"
CDS 2646..4361
/codon_start=1
/product="N-terminal portion of Streptococcus pyogenes Cas9
(Zetsche et al., 2015)"
/label=Cas9(N)
/translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAERTRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
CDS 6777..7025
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
CDS 7056..7304
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
CDS 7347..7367
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7386..7442
/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 7443..8162
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(7488..7509)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(7749..7768)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 8096..8117
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(8249..8268)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_site 8314..8369
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal"
primer_bind complement(8368..8387)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(8730..8746)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8730..8746)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(8743..8765)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 8754..8770
/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(8778..8808)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8823..8844
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 8903..9098
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 8949..9084
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 9011..9030
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 9104..9238
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(9141..9160)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 9195..9214
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(9306..9323)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(9477..10065)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(9557..9576)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(10236..11096)
/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 10859..10878
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(11097..6)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 tccgcgcaca tttccccgaa aagtgccacc tgggtcgaca ttgattattg actagttatt
61 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
121 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
181 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
241 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
301 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
361 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg
421 aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt
481 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggcgc
541 gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg
601 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg
661 cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc
721 gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc
781 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa
841 tgacggcttg tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagggccct
901 ttgtgcgggg ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg
961 tgcggctccg cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg
1021 ctccgcagtg tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg cggggggggc
1081 tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg
1141 ggcgcgtcgg tcgggctgca accccccctg cacccccctc cccgagttgc tgagcacggc
1201 ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg
1261 gggtggcggc aggtgggggt gccgggcggg gcggggccgc ctcgggccgg ggagggctcg
1321 ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc
1381 cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt
1441 gcggagccga aatctgggag gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg
1501 tgcggcgccg gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt
1561 ccccttctcc ctctccagcc tcggggctgt ccgcgggggg acggctgcct tcggggggga
1621 cggggcaggg cggggttcgg cttctggcgt gtgaccggcg gctctagagc ctctgctaac
1681 catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg
1741 tctcatcatt ttggcaaaga attctgcagt cgacggtacc gcgggcccgg gatccaccgg
1801 tcgccaccat gaaacggaca gccgacggaa gcgagttcga gtcaccaaag aagaagcgga
1861 aagtctcctc agagactggg cctgtcgccg tcgatccaac cctgcgccgc cggattgaac
1921 ctcacgagtt tgaagtgttc tttgaccccc gggagctgag aaaggagaca tgcctgctgt
1981 acgagatcaa ctggggaggc aggcactcca tctggaggca cacctctcag aacacaaata
2041 agcacgtgga ggtgaacttc atcgagaagt ttaccacaga gcggtacttc tgccccaata
2101 ccagatgtag catcacatgg tttctgagct ggtccccttg cggagagtgt agcagggcca
2161 tcaccgagtt cctgtccaga tatccacacg tgacactgtt tatctacatc gccaggctgt
2221 atcaccacgc agacccaagg aataggcagg gcctgcgcga tctgatcagc tccggcgtga
2281 ccatccagat catgacagag caggagtccg gctactgctg gcggaacttc gtgaattatt
2341 ctcctagcaa cgaggcccac tggcctaggt acccacacct gtgggtgcgc ctgtacgtgc
2401 tggagctgta ttgcatcatc ctgggcctgc ccccttgtct gaatatcctg cggagaaagc
2461 agccccagct gaccttcttt acaatcgccc tgcagtcttg tcactatcag aggctgccac
2521 cccacatcct gtgggccaca ggcctgaagt ctggaggatc tagcggagga tcctctggca
2581 gcgagacacc aggaacaagc gagtcagcaa caccagagag cagtggcggc agcagcggcg
2641 gcagcgacaa gaagtacagc atcggcctgg ccatcggcac caactctgtg ggctgggccg
2701 tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc
2761 ggcacagcat caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg
2821 agagaacccg gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct
2881 gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca
2941 gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg
3001 gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa
3061 agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc
3121 acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg
3181 acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc
3241 ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca
3301 gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa
3361 acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg
3421 aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg
3481 cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca
3541 tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct
3601 ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc
3661 ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg
3721 ccggctacat tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc
3781 tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc
3841 ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc
3901 acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga
3961 tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca
4021 gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg
4081 aagtggtgga caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata
4141 agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg
4201 tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc
4261 tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga
4321 ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa
4381 tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa
4441 ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg
4501 tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg
4561 cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg
4621 gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc
4681 tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg
4741 acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc
4801 tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga
4861 cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg
4921 tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga
4981 gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc
5041 ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc
5101 gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacc
5161 atatcgtgcc tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa
5221 gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga
5281 agaactactg gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc
5341 tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac
5401 agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga
5461 acactaagta cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt
5521 ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca
5581 actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa
5641 agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga
5701 agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca
5761 gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc
5821 ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt
5881 ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg
5941 tgcagacagg cggcttcagc aaagagtcta tcagacccaa gaggaacagc gataagctga
6001 tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcctgtgg cccaccgtgg
6061 cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg
6121 tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg
6181 actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta
6241 agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct gccaagcagc
6301 tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca
6361 gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg
6421 aacagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag
6481 tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata
6541 agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc agactgggag
6601 cccctagagc cttcaagtac tttgacacca ccatcgaccc caagcagtac agaagcacca
6661 aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga
6721 tcgacctgtc tcagctggga ggtgacagcg gcgggagcgg cgggagcggg gggagcacta
6781 atctgagcga catcattgag aaggagactg ggaaacagct ggtcattcag gagtccatcc
6841 tgatgctgcc tgaggaggtg gaggaagtga tcggcaacaa gccagagtct gacatcctgg
6901 tgcacaccgc ctacgacgag tccacagatg agaatgtgat gctgctgacc tctgacgccc
6961 ccgagtataa gccttgggcc ctggtcatcc aggattctaa cggcgagaat aagatcaaga
7021 tgctgagcgg aggatccgga ggatctggag gcagcaccaa cctgtctgac atcatcgaga
7081 aggagacagg caagcagctg gtcatccagg agagcatcct gatgctgccc gaagaagtcg
7141 aagaagtgat cggaaacaag cctgagagcg atatcctggt ccataccgcc tacgacgaga
7201 gtaccgacga aaatgtgatg ctgctgacat ccgacgcccc agagtataag ccctgggctc
7261 tggtcatcca ggattccaac ggagagaaca aaatcaaaat gctgtctggc ggctcaaaaa
7321 gaaccgccga cggcagcgaa ttcgagccca agaagaagag gaaagtcgga ggcggtggaa
7381 gcggcgcaac aaacttctct ctgctgaaac aagccggaga tgtcgaagag aatcctggac
7441 cgatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
7501 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
7561 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
7621 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
7681 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
7741 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
7801 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
7861 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
7921 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
7981 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
8041 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
8101 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
8161 gaagcggccg cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca
8221 atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca
8281 tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa
8341 tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt
8401 taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg
8461 ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc
8521 attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg
8581 tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt
8641 tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg agatccctcg
8701 acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
8761 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
8821 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
8881 aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc ccgcccctaa
8941 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
9001 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
9061 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc
9121 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
9181 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccg
9241 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
9301 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
9361 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
9421 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
9481 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
9541 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
9601 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
9661 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
9721 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
9781 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
9841 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
9901 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
9961 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
10021 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
10081 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
10141 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
10201 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
10261 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
10321 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
10381 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
10441 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
10501 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
10561 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
10621 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
10681 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
10741 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
10801 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
10861 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
10921 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
10981 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
11041 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
11101 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
11161 tttgaatgta tttagaaaaa taaacaaata ggggt
//
