| 质粒名称: | CGBE1-NG (pBM1246) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 140256 |
| 存储实验室: | Keith Joung |
|
载体骨架基本信息 |
|
| 载体名称: | CMV |
| 载体出品公司: | pCMV-ABEmax-P2A-EGFP (Addgene #112101) |
| 空载体大小: | 3400 bp |
| 完整质粒大小: | 9909 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素100 μg/mL |
| 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: | eUNG-BE4max(R33A)_NG-ΔUGI,bpNLS-UNG(E.coli)-rAPOBEC1(R33A)-nCas9_NG-bpNLS-P2A-EGFP |
| 物种来源: | R. norvegicus (rat); E. coli, S. pyogenes |
| 插入大小: | 6544 bp |
| 基因突变: | R33A in rAPOBEC1, NG mutations in SpCas9, D10A in Cas9 |
| 启动子: | CMV |
| 克隆方法: | Gibson克隆 |
| 5’测序引物 | CGCAAATGGGCGGTAGGCGTG |
| 3’测序引物 | TAGAAGGCACAGTCGAGG |
|
文献引用方法 |
|
| 材料和方法部分: | CGBE1-NG (pBM1246) was a gift from Keith Joung (Addgene plasmid # 140256 ; http://n2t.net/addgene:140256 ; RRID:Addgene_140256) |
| 参考文献部分: | CRISPR C-to-G base editors for inducing targeted DNA transversions in human cells. Kurt IC, Zhou R, Iyer S, Garcia SP, Miller BR, Langner LM, Grunewald J, Joung JK. Nat Biotechnol. 2020 Jul 20. pii: 10.1038/s41587-020-0609-x. doi: 10.1038/s41587-020-0609-x. 10.1038/s41587-020-0609-x PubMed 32690971 |
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 CGBE1-NG (pBM1246) 9909 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION CMV promoter expression plasmid for
UNG(E.coli)-rAPOBEC1(R33A)-nCas9_NG-P2A-EGFP.
ACCESSION .
VERSION .
KEYWORDS CGBE1-NG (pBM1246)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9909)
AUTHORS Kurt IC, Zhou R, Iyer S, Garcia SP, Miller BR, Langner LM, Grunewald
J, Joung JK
TITLE CRISPR C-to-G base editors for inducing targeted DNA transversions
in human cells.
JOURNAL Nat Biotechnol. 2020 Jul 20. pii: 10.1038/s41587-020-0609-x. doi:
10.1038/s41587-020-0609-x.
PUBMED 32690971
REFERENCE 2 (bases 1 to 9909)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9909
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 6..23
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(46..63)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 52..276
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(347..363)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(347..363)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(360..382)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 371..387
/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(395..425)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 440..461
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(578..595)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(749..1337)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(829..848)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1508..2368)
/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 2131..2150
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2369..2473)
/gene="bla"
/label=AmpR promoter
primer_bind complement(2552..2571)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 2743..3122
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 3123..3326
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 3277..3297
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 3368..3387
/label=T7
/note="T7 promoter, forward primer"
promoter 3368..3386
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 3436..3456
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 4951..6666
/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 9094..9114
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 9133..9189
/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 9190..9906
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(9235..9256)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(9496..9515)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 9843..9864
/label=EGFP-C
/note="EGFP, forward primer"
ORIGIN
1 ccggtcatca tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt
61 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
121 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
181 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
241 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
301 ggggctcgat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
361 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
421 gtaaagccta gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
481 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
541 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
601 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
661 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
721 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
781 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
841 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
901 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
961 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
1021 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
1081 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
1141 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
1201 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
1261 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
1321 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac actcagtgga
1381 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
1441 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
1501 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
1561 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
1621 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
1681 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
1741 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
1801 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
1861 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
1921 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
1981 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
2041 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
2101 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
2161 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
2221 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
2281 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
2341 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
2401 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
2461 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
2521 gatctcccga tcccctaggg tcgactctca gtacaatctg ctctgatgcc gcatagttaa
2581 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
2641 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
2701 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta
2761 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
2821 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
2881 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
2941 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
3001 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
3061 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
3121 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
3181 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
3241 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
3301 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagagatccg
3361 cggccgctaa tacgactcac tatagggaga gccgccacca tgaaacggac agccgacgga
3421 agcgagttcg agtcaccaaa gaagaagcgg aaagtcgcca acgaactgac ctggcacgac
3481 gtgctggccg aagagaagca gcagccctat ttccttaaca ccctgcagac cgtggccagc
3541 gagcggcaga gcggcgtgac catctacccc ccacagaaag acgtgtttaa cgccttccgc
3601 ttcacagagc tgggcgacgt gaaggtggtg atcctgggcc aggaccccta ccacggcccc
3661 ggccaggccc atggtctggc cttcagcgtg cggcccggca tcgccatccc ccccagcctg
3721 ctgaacatgt ataaggagct ggaaaacacc atccccggct tcacccggcc caatcacggc
3781 tacctggaga gctgggcgcg gcagggcgtg ctgctgctca acaccgtgct gacggtacgg
3841 gccggccagg cgcatagcca cgccagcctg ggctgggaga ccttcaccga taaggtgatc
3901 agcctgatca accagcaccg ggagggcgtg gtgtttctgc tgtggggcag ccatgcccag
3961 aagaaaggcg ccatcatcga caagcagcgg catcatgtgc tgaaagcccc ccaccccagc
4021 ccccttagcg cccaccgggg cttcttcggc tgcaaccatt tcgtgctggc caatcagtgg
4081 ctggaacaac ggggcgagac ccccattgac tggatgcccg tgttgcccgc cgagagcgag
4141 agcggcggga gcggcgggag cggggggagc tcctcagaga ctgggcctgt cgccgtcgat
4201 ccaaccctgc gccgccggat tgaacctcac gagtttgaag tgttctttga cccccgggag
4261 ctggccaagg agacatgcct gctgtacgag atcaactggg gaggcaggca ctccatctgg
4321 aggcacacct ctcagaacac aaataagcac gtggaggtga acttcatcga gaagtttacc
4381 acagagcggt acttctgccc caataccaga tgtagcatca catggtttct gagctggtcc
4441 ccttgcggag agtgtagcag ggccatcacc gagttcctgt ccagatatcc acacgtgaca
4501 ctgtttatct acatcgccag gctgtatcac cacgcagacc caaggaatag gcagggcctg
4561 cgcgatctga tcagctccgg cgtgaccatc cagatcatga cagagcagga gtccggctac
4621 tgctggcgga acttcgtgaa ttattctcct agcaacgagg cccactggcc taggtaccca
4681 cacctgtggg tgcgcctgta cgtgctggag ctgtattgca tcatcctggg cctgccccct
4741 tgtctgaata tcctgcggag aaagcagccc cagctgacct tctttacaat cgccctgcag
4801 tcttgtcact atcagaggct gccaccccac atcctgtggg ccacaggcct gaagtctgga
4861 ggatctagcg gaggatcctc tggcagcgag acaccaggaa caagcgagtc agcaacacca
4921 gagagcagtg gcggcagcag cggcggcagc gacaagaagt acagcatcgg cctggccatc
4981 ggcaccaact ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa
5041 ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga agaacctgat cggagccctg
5101 ctgttcgaca gcggcgaaac agccgaggcc acccggctga agagaaccgc cagaagaaga
5161 tacaccagac ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa cgagatggcc
5221 aaggtggacg acagcttctt ccacagactg gaagagtcct tcctggtgga agaggataag
5281 aagcacgagc ggcaccccat cttcggcaac atcgtggacg aggtggccta ccacgagaag
5341 taccccacca tctaccacct gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg
5401 cggctgatct atctggccct ggcccacatg atcaagttcc ggggccactt cctgatcgag
5461 ggcgacctga accccgacaa cagcgacgtg gacaagctgt tcatccagct ggtgcagacc
5521 tacaaccagc tgttcgagga aaaccccatc aacgccagcg gcgtggacgc caaggccatc
5581 ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc tgatcgccca gctgcccggc
5641 gagaagaaga atggcctgtt cggaaacctg attgccctga gcctgggcct gacccccaac
5701 ttcaagagca acttcgacct ggccgaggat gccaaactgc agctgagcaa ggacacctac
5761 gacgacgacc tggacaacct gctggcccag atcggcgacc agtacgccga cctgtttctg
5821 gccgccaaga acctgtccga cgccatcctg ctgagcgaca tcctgagagt gaacaccgag
5881 atcaccaagg cccccctgag cgcctctatg atcaagagat acgacgagca ccaccaggac
5941 ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg agaagtacaa agagattttc
6001 ttcgaccaga gcaagaacgg ctacgccggc tacattgacg gcggagccag ccaggaagag
6061 ttctacaagt tcatcaagcc catcctggaa aagatggacg gcaccgagga actgctcgtg
6121 aagctgaaca gagaggacct gctgcggaag cagcggacct tcgacaacgg cagcatcccc
6181 caccagatcc acctgggaga gctgcacgcc attctgcggc ggcaggaaga tttttaccca
6241 ttcctgaagg acaaccggga aaagatcgag aagatcctga ccttccgcat cccctactac
6301 gtgggccctc tggccagggg aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa
6361 accatcaccc cctggaactt cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc
6421 atcgagcgga tgaccaactt cgataagaac ctgcccaacg agaaggtgct gcccaagcac
6481 agcctgctgt acgagtactt caccgtgtat aacgagctga ccaaagtgaa atacgtgacc
6541 gagggaatga gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat cgtggacctg
6601 ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga aagaggacta cttcaagaaa
6661 atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg
6721 ggcacatacc acgatctgct gaaaattatc aaggacaagg acttcctgga caatgaggaa
6781 aacgaggaca ttctggaaga tatcgtgctg accctgacac tgtttgagga cagagagatg
6841 atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat gaagcagctg
6901 aagcggcgga gatacaccgg ctggggcagg ctgagccgga agctgatcaa cggcatccgg
6961 gacaagcagt ccggcaagac aatcctggat ttcctgaagt ccgacggctt cgccaacaga
7021 aacttcatgc agctgatcca cgacgacagc ctgaccttta aagaggacat ccagaaagcc
7081 caggtgtccg gccagggcga tagcctgcac gagcacattg ccaatctggc cggcagcccc
7141 gccattaaga agggcatcct gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg
7201 ggccggcaca agcccgagaa catcgtgatc gaaatggcca gagagaacca gaccacccag
7261 aagggacaga agaacagccg cgagagaatg aagcggatcg aagagggcat caaagagctg
7321 ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa cgagaagctg
7381 tacctgtact acctgcagaa tgggcgggat atgtacgtgg accaggaact ggacatcaac
7441 cggctgtccg actacgatgt ggaccatatc gtgcctcaga gctttctgaa ggacgactcc
7501 atcgacaaca aggtgctgac cagaagcgac aagaaccggg gcaagagcga caacgtgccc
7561 tccgaagagg tcgtgaagaa gatgaagaac tactggcggc agctgctgaa cgccaagctg
7621 attacccaga gaaagttcga caatctgacc aaggccgaga gaggcggcct gagcgaactg
7681 gataaggccg gcttcatcaa gagacagctg gtggaaaccc ggcagatcac aaagcacgtg
7741 gcacagatcc tggactcccg gatgaacact aagtacgacg agaatgacaa gctgatccgg
7801 gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg atttccggaa ggatttccag
7861 ttttacaaag tgcgcgagat caacaactac caccacgccc acgacgccta cctgaacgcc
7921 gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc
7981 gactacaagg tgtacgacgt gcggaagatg atcgccaaga gcgagcagga aatcggcaag
8041 gctaccgcca agtacttctt ctacagcaac atcatgaact ttttcaagac cgagattacc
8101 ctggccaacg gcgagatccg gaagcggcct ctgatcgaga caaacggcga aaccggggag
8161 atcgtgtggg ataagggccg ggattttgcc accgtgcgga aagtgctgag catgccccaa
8221 gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga gtctatccgg
8281 cccaagagga acagcgataa gctgatcgcc agaaagaagg actgggaccc taagaagtac
8341 ggcggcttcg tgagccccac cgtggcctat tctgtgctgg tggtggccaa agtggaaaag
8401 ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat catggaaaga
8461 agcagcttcg agaagaatcc catcgacttt ctggaagcca agggctacaa agaagtgaaa
8521 aaggacctga tcatcaagct gcctaagtac tccctgttcg agctggaaaa cggccggaag
8581 agaatgctgg cctctgcccg gttcctgcag aagggaaacg aactggccct gccctccaaa
8641 tatgtgaact tcctgtacct ggccagccac tatgagaagc tgaagggctc ccccgaggat
8701 aatgagcaga aacagctgtt tgtggaacag cacaagcact acctggacga gatcatcgag
8761 cagatcagcg agttctccaa gagagtgatc ctggccgacg ctaatctgga caaagtgctg
8821 tccgcctaca acaagcaccg ggataagccc atcagagagc aggccgagaa tatcatccac
8881 ctgtttaccc tgaccaatct gggagcccct cgggccttca agtactttga caccaccatc
8941 gaccggaagg tgtaccggag caccaaagag gtgctggacg ccaccctgat ccaccagagc
9001 atcaccggcc tgtacgagac acggatcgac ctgtctcagc tgggaggtga ctctggcggc
9061 tcaaaaagaa ccgccgacgg cagcgaattc gagcccaaga agaagaggaa agtcggaggc
9121 ggtggaagcg gcgcaacaaa cttctctctg ctgaaacaag ccggagatgt cgaagagaat
9181 cctggaccga tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
9241 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
9301 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
9361 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
9421 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
9481 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
9541 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
9601 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
9661 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
9721 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
9781 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
9841 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
9901 tacaagtga
//
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