| 质粒名称: | pCMV-T7-ABEmax(7.10)-SpRY-P2A-EGFP (RTW5025) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 140003 |
| 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 |
| 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 ABEmax(7.10) SpCas9 variant named SpRY with P2A-EGFP,RTW5025 |
| 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 | CMV and T7 |
| 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 | oBK6928-CCAAGTCTCCACCCCATTGACG |
| 3' 测序引物 | oBK219-GGGAGTGGCACCTTCCAGGGTC |
|
文献引用方法 |
|
| 材料和方法部分: | pCMV-T7-ABEmax(7.10)-SpRY-P2A-EGFP (RTW5025) was a gift from Benjamin Kleinstiver (Addgene plasmid # 140003 ; http://n2t.net/addgene:140003 ; RRID:Addgene_140003) |
| 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
-SpRY-P2A-EGFP (RTW5025)质粒图谱.png)
载体简介
载体序列
LOCUS pCMV-T7-ABEmax(7.10)-SpRY-P2A-EGFP (RTW5025) 9627 bp ds-DNA circular SYN 20-MAR-2020
DEFINITION CMV and T7 promoter expression plasmid for human codon optimized
ABEmax(7.10) A-to-G base editor with
SpRY(D10A/A61R/L1111R/D1135L/S1136W/G1218K/E1219Q/N1317R/A1322R/R133
3P/R1335Q/T1337R) and P2A-EGFP.
ACCESSION .
VERSION .
KEYWORDS pCMV-T7-ABEmax(7.10)-SpRY-P2A-EGFP (RTW5025)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9627)
AUTHORS Kleinstiver
TITLE Kleinstiver lab CRISPR plasmids
JOURNAL unpublished
REFERENCE 2 (bases 1 to 9627)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Mar 20, 2020 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9627
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 511..530
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(749..853)
/gene="bla"
/label=AmpR promoter
primer_bind complement(932..951)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 1123..1502
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 1503..1706
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 1657..1677
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 1748..1767
/label=T7
/note="T7 promoter, forward primer"
promoter 1748..1766
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1816..1836
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 3025..4740
/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 7168..7188
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7198..7254
/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 7255..7971
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(7300..7321)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(7561..7580)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 7908..7929
/label=EGFP-C
/note="EGFP, forward primer"
CDS 7984..8004
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 8013..8030
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(8053..8070)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 8059..8283
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(8354..8370)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8354..8370)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(8367..8389)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 8378..8394
/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(8402..8432)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8447..8468
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(8585..8602)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(8756..9344)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(8836..8855)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(9515..748)
/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 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
61 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
121 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
181 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
241 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
301 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
361 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
421 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
481 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
541 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
601 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
661 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
721 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
781 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
841 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
901 gatctcccga tcccctaggg tcgactctca gtacaatctg ctctgatgcc gcatagttaa
961 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
1021 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
1081 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta
1141 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
1201 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
1261 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
1321 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
1381 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
1441 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
1501 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
1561 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
1621 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
1681 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagagatccg
1741 cggccgctaa tacgactcac tatagggaga gccgccacca tgaaacggac agccgacgga
1801 agcgagttcg agtcaccaaa gaagaagcgg aaagtctctg aagtcgagtt tagccacgag
1861 tattggatga ggcacgcact gaccctggca aagcgagcat gggatgaaag agaagtcccc
1921 gtgggcgccg tgctggtgca caacaataga gtgatcggag agggatggaa caggccaatc
1981 ggccgccacg accctaccgc acacgcagag atcatggcac tgaggcaggg aggcctggtc
2041 atgcagaatt accgcctgat cgatgccacc ctgtatgtga cactggagcc atgcgtgatg
2101 tgcgcaggag caatgatcca cagcaggatc ggaagagtgg tgttcggagc acgggacgcc
2161 aagaccggcg cagcaggctc cctgatggat gtgctgcacc accccggcat gaaccaccgg
2221 gtggagatca cagagggaat cctggcagac gagtgcgccg ccctgctgag cgatttcttt
2281 agaatgcgga gacaggagat caaggcccag aagaaggcac agagctccac cgactctgga
2341 ggatctagcg gaggatcctc tggaagcgag acaccaggca caagcgagtc cgccacacca
2401 gagagctccg gcggctcctc cggaggatcc tctgaggtgg agttttccca cgagtactgg
2461 atgagacatg ccctgaccct ggccaagagg gcacgcgatg agagggaggt gcctgtggga
2521 gccgtgctgg tgctgaacaa tagagtgatc ggcgagggct ggaacagagc catcggcctg
2581 cacgacccaa cagcccatgc cgaaattatg gccctgagac agggcggcct ggtcatgcag
2641 aactacagac tgattgacgc caccctgtac gtgacattcg agccttgcgt gatgtgcgcc
2701 ggcgccatga tccactctag gatcggccgc gtggtgtttg gcgtgaggaa cgcaaaaacc
2761 ggcgccgcag gctccctgat ggacgtgctg cactaccccg gcatgaatca ccgcgtcgaa
2821 attaccgagg gaatcctggc agatgaatgt gccgccctgc tgtgctattt ctttcggatg
2881 cctagacagg tgttcaatgc tcagaagaag gcccagagct ccaccgactc cggaggatct
2941 agcggaggct cctctggctc tgagacacct ggcacaagcg agagcgcaac acctgaaagc
3001 agcgggggca gcagcggggg gtcagacaag aagtacagca tcggcctggc catcggcacc
3061 aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag
3121 gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc
3181 gacagcggcg aaacagccga gagaacccgg ctgaagagaa ccgccagaag aagatacacc
3241 agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg
3301 gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac
3361 gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc
3421 accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg
3481 atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac
3541 ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac
3601 cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct
3661 gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag
3721 aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag
3781 agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac
3841 gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc
3901 aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc
3961 aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc
4021 ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac
4081 cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga agagttctac
4141 aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg
4201 aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag
4261 atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg
4321 aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc
4381 cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc
4441 accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag
4501 cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg
4561 ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga
4621 atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc
4681 aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag
4741 tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca
4801 taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag
4861 gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag
4921 gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg
4981 cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag
5041 cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc
5101 atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg
5161 tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt
5221 aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg
5281 cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga
5341 cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc
5401 cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg
5461 tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg
5521 tccgactacg atgtggacca tatcgtgcct cagagctttc tgaaggacga ctccatcgac
5581 aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa
5641 gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc
5701 cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag
5761 gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag
5821 atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg
5881 aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac
5941 aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg
6001 ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac
6061 aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc
6121 gccaagtact tcttctacag caacatcatg aactttttca agaccgagat taccctggcc
6181 aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg
6241 tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat
6301 atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat cagacccaag
6361 aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc
6421 ttcctgtggc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag
6481 tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc
6541 ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac
6601 ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg
6661 ctggcctctg ccaagcagct gcagaaggga aacgaactgg ccctgccctc caaatatgtg
6721 aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag
6781 cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc
6841 agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc
6901 tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt
6961 accctgacca gactgggagc ccctagagcc ttcaagtact ttgacaccac catcgacccc
7021 aagcagtaca gaagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc
7081 ggcctgtacg agacacggat cgacctgtct cagctgggag gtgactctgg cggctcaaaa
7141 agaaccgccg acggcagcga attcgagccc aagaagaaga ggaaagtcgg aagcggagct
7201 actaacttca gcctgctgaa gcaggctgga gacgtggagg agaaccctgg acctatggtg
7261 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
7321 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
7381 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
7441 accaccctga cctatggagt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
7501 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
7561 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
7621 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
7681 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
7741 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
7801 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
7861 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
7921 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtctggtggt
7981 tctcccaaga agaagaggaa agtctaaccg gtcatcatca ccatcaccat tgagtttaaa
8041 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
8101 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
8161 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
8221 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
8281 tggcttctga ggcggaaaga accagctggg gctcgatacc gtcgacctct agctagagct
8341 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
8401 acaacatacg agccggaagc ataaagtgta aagcctaggg tgcctaatga gtgagctaac
8461 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
8521 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
8581 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
8641 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
8701 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
8761 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
8821 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
8881 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
8941 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
9001 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
9061 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
9121 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
9181 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
9241 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
9301 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
9361 tttctacggg gtctgacact cagtggaacg aaaactcacg ttaagggatt ttggtcatga
9421 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
9481 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
9541 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
9601 taactacgat acgggagggc ttaccat
//
