| 质粒名称: | SPACE (pRZ1813) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 140242 |
| 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: | 10308 bp |
| 载体类型: | 哺乳细胞表达 |
| 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: | SPACE,bpNLS-TadA*(V82G)-nCas9-bpNLS-pmCDA1(R187W)-UGI-UGI-P2A-EGFP |
| 物种来源: | E. coli, S. pyogenes, P. marinus |
| 插入大小: | 6943 bp |
| 基因突变: | V82G in TadA*, D10A in Cas9, R187W in pmCDA1 |
| 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: | SPACE (pRZ1813) was a gift from Keith Joung (Addgene plasmid # 140242 ; http://n2t.net/addgene:140242 ; RRID:Addgene_140242) |
| 参考文献部分: | A dual-deaminase CRISPR base editor enables concurrent adenine and cytosine editing. Grunewald J, Zhou R, Lareau CA, Garcia SP, Iyer S, Miller BR, Langner LM, Hsu JY, Aryee MJ, Joung JK. Nat Biotechnol. 2020 Jun 1. pii: 10.1038/s41587-020-0535-y. doi: 10.1038/s41587-020-0535-y. 10.1038/s41587-020-0535-y PubMed 32483364 |
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 SPACE (pRZ1813) 10308 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION CMV promoter expression plasmid for
TadA*(V82G)-nCas9-pmCDA1(R187W)-UGI-UGI-P2A-EGFP.
ACCESSION .
VERSION .
KEYWORDS SPACE (pRZ1813)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10308)
AUTHORS Grunewald J, Zhou R, Lareau CA, Garcia SP, Iyer S, Miller BR,
Langner LM, Hsu JY, Aryee MJ, Joung JK
TITLE A dual-deaminase CRISPR base editor enables concurrent adenine and
cytosine editing.
JOURNAL Nat Biotechnol. 2020 Jun 1. pii: 10.1038/s41587-020-0535-y. doi:
10.1038/s41587-020-0535-y.
PUBMED 32483364
REFERENCE 2 (bases 1 to 10308)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..10308
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 626..645
/label=T7
/note="T7 promoter, forward primer"
promoter 626..644
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 694..714
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1309..5409
/codon_start=1
/product="nickase mutant of the Cas9 endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system"
/label=Cas9(D10A)
/note="generates RNA-guided single strand nicks in DNA due
to the D10A mutation in the RuvC catalytic domain"
/translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 5452..5472
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 6220..6468
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
CDS 6499..6747
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
CDS 6757..6813
/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 6814..7530
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(6859..6880)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(7120..7139)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 7467..7488
/label=EGFP-C
/note="EGFP, forward primer"
CDS 7543..7563
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7572..7589
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(7612..7629)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 7618..7842
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(7913..7929)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7913..7929)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(7926..7948)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 7937..7953
/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(7961..7991)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8006..8027
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(8144..8161)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(8315..8903)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(8395..8414)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(9074..9934)
/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 9697..9716
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(9935..10039)
/gene="bla"
/label=AmpR promoter
primer_bind complement(10118..10137)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
601 agatccgcta gagatccgcg gccgctaata cgactcacta tagggagagc cgccaccatg
661 aaacggacag ccgacggaag cgagttcgag tcaccaaaga agaagcggaa agtctctgag
721 gtggagtttt cccacgagta ctggatgaga catgccctga ccctggccaa gagggcacgc
781 gatgagaggg aggtgcctgt gggagccgtg ctggtgctga acaatagagt gatcggcgag
841 ggctggaaca gagccatcgg cctgcacgac ccaacagccc atgccgaaat tatggccctg
901 agacagggcg gcctggtcat gcagaactac agactgattg acgccaccct gtacggcaca
961 ttcgagcctt gcgtgatgtg cgccggcgcc atgatccact ctaggatcgg ccgcgtggtg
1021 tttggcgtga ggaacgcaaa aaccggcgcc gcaggctccc tgatggacgt gctgcactac
1081 cccggcatga atcaccgcgt cgaaattacc gagggaatcc tggcagatga atgtgccgcc
1141 ctgctgtgct atttctttcg gatgcctaga caggtgttca atgctcagaa gaaggcccag
1201 agctccaccg actccggagg atctagcgga ggctcctctg gctctgagac acctggcaca
1261 agcgagagcg caacacctga aagcagcggg ggcagcagcg gggggtcaga caagaagtac
1321 agcatcggcc tggccatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac
1381 aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag
1441 aacctgatcg gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag
1501 agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc
1561 ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc
1621 ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag
1681 gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc
1741 accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg
1801 ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc
1861 atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc
1921 gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg
1981 atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc
2041 ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag
2101 ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag
2161 tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc
2221 ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac
2281 gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag
2341 aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc
2401 ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc
2461 accgaggaac tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc
2521 gacaacggca gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg
2581 caggaagatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc
2641 ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg
2701 accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc
2761 gcttccgccc agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag
2821 aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc
2881 aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa
2941 aaggccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa
3001 gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat
3061 cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac
3121 ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg
3181 tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac
3241 aaagtgatga agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag
3301 ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc
3361 gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa
3421 gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc
3481 aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac
3541 gagctcgtga aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga
3601 gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa
3661 gagggcatca aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag
3721 ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac
3781 caggaactgg acatcaaccg gctgtccgac tacgatgtgg accatatcgt gcctcagagc
3841 tttctgaagg acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc
3901 aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag
3961 ctgctgaacg ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga
4021 ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg
4081 cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag
4141 aatgacaagc tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat
4201 ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac
4261 gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa
4321 agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc
4381 gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt
4441 ttcaagaccg agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca
4501 aacggcgaaa ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa
4561 gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc
4621 agcaaagagt ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac
4681 tgggacccta agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg
4741 gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg
4801 atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag
4861 ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag
4921 ctggaaaacg gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa
4981 ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg
5041 aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac
5101 ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct
5161 aatctggaca aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag
5221 gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag
5281 tactttgaca ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc
5341 accctgatcc accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg
5401 ggaggtgact ctggcggctc aaaaagaacc gccgacggca gcgaattcga gcccaagaag
5461 aagaggaaag tctctggagg atctagcgga ggatcctctg gaagcgagac accaggcaca
5521 agcgagtccg ccacaccaga gagctccggc ggctcctccg gaggatccac agacgccgag
5581 tacgtgcgca ttcatgagaa actggatatt tacaccttca agaagcagtt cttcaacaac
5641 aagaaatctg tgtcacaccg ctgctacgtg ctgtttgagt tgaagcgaag gggcgaaaga
5701 agggcttgct tttggggcta tgccgtcaac aagccccaaa gtggcaccga gagaggaata
5761 cacgctgaga tattcagtat ccgaaaggtg gaagagtatc ttcgggataa tcctgggcag
5821 tttacgatca actggtattc cagctggagt ccttgcgctg attgtgccga gaaaattctg
5881 gaatggtata atcaggaact tcggggaaac gggcacacat tgaaaatctg ggcctgcaag
5941 ctgtactacg agaagaatgc ccggaaccag ataggactct ggaatctgag ggacaatggt
6001 gtaggcctga acgtgatggt ttccgagcac tatcagtgtt gtcggaagat tttcatccaa
6061 agctctcata accagctcaa tgaaaaccgc tggttggaga aaacactgaa acgtgcggag
6121 aagtggagat ccgagctgag catcatgatc caggtcaaga ttctgcatac cactaagtct
6181 ccagccgtta gcggcgggag cggcgggagc ggggggagca ctaatctgag cgacatcatt
6241 gagaaggaga ctgggaaaca gctggtcatt caggagtcca tcctgatgct gcctgaggag
6301 gtggaggaag tgatcggcaa caagccagag tctgacatcc tggtgcacac cgcctacgac
6361 gagtccacag atgagaatgt gatgctgctg acctctgacg cccccgagta taagccttgg
6421 gccctggtca tccaggattc taacggcgag aataagatca agatgctgag cggaggatcc
6481 ggaggatctg gaggcagcac caacctgtct gacatcatcg agaaggagac aggcaagcag
6541 ctggtcatcc aggagagcat cctgatgctg cccgaagaag tcgaagaagt gatcggaaac
6601 aagcctgaga gcgatatcct ggtccatacc gcctacgacg agagtaccga cgaaaatgtg
6661 atgctgctga catccgacgc cccagagtat aagccctggg ctctggtcat ccaggattcc
6721 aacggagaga acaaaatcaa aatgctggga agcggagcta ctaacttcag cctgctgaag
6781 caggctggag acgtggagga gaaccctgga cctatggtga gcaagggcga ggagctgttc
6841 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
6901 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
6961 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctatggagtg
7021 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
7081 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
7141 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
7201 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
7261 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
7321 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
7381 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
7441 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
7501 atcactctcg gcatggacga gctgtacaag tctggtggtt ctcccaagaa gaagaggaaa
7561 gtctaaccgg tcatcatcac catcaccatt gagtttaaac ccgctgatca gcctcgactg
7621 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
7681 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
7741 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
7801 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
7861 ccagctgggg ctcgataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
7921 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
7981 taaagtgtaa agcctagggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
8041 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
8101 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
8161 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
8221 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
8281 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
8341 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
8401 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
8461 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
8521 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
8581 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
8641 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
8701 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
8761 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
8821 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
8881 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacactc
8941 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
9001 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
9061 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
9121 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
9181 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
9241 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
9301 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
9361 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
9421 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
9481 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
9541 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
9601 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
9661 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
9721 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
9781 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
9841 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
9901 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
9961 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
10021 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
10081 gagatcgatc tcccgatccc ctagggtcga ctctcagtac aatctgctct gatgccgcat
10141 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
10201 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg
10261 ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgtt
//
