| 质粒名称: | pCRISPR-cBEST |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 125689 |
| 存储实验室: | Tilmann Weber |
|
载体骨架基本信息 |
|
| 载体名称: | pGM1190 |
| 载体出品公司: | -- |
| 载体抗性: | 安普霉素 |
| 空载体大小: | 6971 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | CRISPR，合成生物学 |
| 筛选标记: | -- |
| 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: | streptomyces codon optimized spCas9n (D10A), modified sgRNA casette, streptomyces codon optimized rAPOBEC1,and streptomyces codon optimized UGI |
| 物种来源: | Other |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | ermE*/tipA |
| 标签/融合蛋白: | -- |
| 克隆方法: | Gibson克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pCRISPR-cBEST was a gift from Tilmann Weber (Addgene plasmid # 125689 ; http://n2t.net/addgene:125689 ; RRID:Addgene_125689) |
| 参考文献部分: | Highly efficient DSB-free base editing for streptomycetes with CRISPR-BEST. Tong Y, Whitford CM, Robertsen HL, Blin K, Jorgensen TS, Klitgaard AK, Gren T, Jiang X, Weber T, Lee SY. Proc Natl Acad Sci U S A. 2019 Oct 8;116(41):20366-20375. doi: 10.1073/pnas.1913493116. Epub 2019 Sep 23. 10.1073/pnas.1913493116 PubMed 31548381 |
买家导航
pCRISPRcBEST载体基本信息
pCRISPRcBEST载体图谱质粒图谱和多克隆位点信息
pCRISPRcBEST载体简介
pCRISPRcBEST载体序列
LOCUS pCRISPR-cBEST 12247 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION C to T base editor for actinomycetes.
ACCESSION .
VERSION .
KEYWORDS pCRISPR-cBEST
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12247)
AUTHORS Tong Y, Whitford CM, Robertsen HL, Blin K, Jorgensen TS, Klitgaard
AK, Gren T, Jiang X, Weber T, Lee SY
TITLE Highly efficient DSB-free base editing for streptomycetes with
CRISPR-BEST.
JOURNAL Proc Natl Acad Sci U S A. 2019 Oct 8;116(41):20366-20375. doi:
10.1073/pnas.1913493116. Epub 2019 Sep 23.
PUBMED 31548381
REFERENCE 2 (bases 1 to 12247)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..12247
/organism="synthetic DNA construct"
/mol_type="other DNA"
oriT 466..575
/note="incP origin of transfer"
CDS 891..1691
/codon_start=1
/gene="aac(3)IV"
/product="aminoglycoside 3-N-acetyltransferase type IV
"
/label=ApmR
/note="confers resistance to apramycin (Paget and Davies,
1996)"
/translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDSFWRLPNV
KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
LAELLAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
rep_origin 1993..2580
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2481..2500
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2734..2751
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 2868..2889
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2904..2934
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2942..2958
/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)."
primer_bind 2947..2969
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 2966..2982
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2966..2982
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
misc_RNA 3119..3194
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
terminator 3364..3398
/label=lambda t0 terminator
/note="minimal transcription terminator from phage lambda
(Scholtissek and Grosse, 1987)"
CDS 3566..4252
/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="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
LWATGLK"
CDS 4301..8401
/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 8414..8665
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
terminator 8846..8894
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
CDS 10609..12051
/codon_start=1
/gene="rep"
/product="replication initiator protein from the
Streptomyces ghanaensis plasmid pSG5"
/label=pSG5 Rep
/translation="MWKPGEATWGNTCRCNNVHTCPWCMSRILAVRGSNVQLAADGLAD
AGYGLHLGTNTLRHFERMAFGTVRKGMRHGLVAVLHDGWKGAYGSSGRRWRTMRDDFGI
IGYERAFEDTFGWGSGWHLHWHTLWVTREVLGPDAQAAFRDALAGAWAAGVESAGGYTV
SETCDRPGCSCEGKGHGTDVRPLNGADAADGDAGKQARYLYKDGDKTKGGVAKIGLELA
GQNFKAGRGDDRMGPLDLGDAAAAELQRLRRPGPFVEKYREREFGVFQVRKHYRSQNLN
RLIKELGIQQDVRTEEEITDDTEGLVAIAVIPAYIWYRYIARVAGRRLDLIKVAETYGL
PGVRRLVESWGLVWGKDVLDPPAPEAPAAPGDLDADQMRFEVMSEEEAAFREARRKANE
ARTEELAASLDRVRQPKKEAIRPTISLRKRLKPKPVTVDVKTPPPGAASPVCRRCKGKL
APVLQPWGQHPGDCLRVDTAVA"
ORIGIN
1 ggccgtcgac gacgacgcgt ccgcctgcct gcttttggcc ggcgggccgg gccgcggttg
61 tccgccgggt cggaccggtc ccttggtcga ccgatgcctc ggccacgacg atgctgggcc
121 cgaagcggta cgcgacccgt gccgggccgc agcggatcgc ggcagcagga gctccccgcc
181 cgccacggct ggctggatgg tcgcctcacg cgcccgcagg ggtgacggcg tgggtgcggc
241 ggtcagcccc gtgatcgggc gtcagacggc cgctgaggga ccgcgaccac ccctcccttc
301 ccgcctgccg tctttcaccg cccggccggt ccgcagacgc cgtacggcca ggccacggag
361 gattacgggc agatggcgcc cacccggcac ccgggctggg gcaccctgga cagcacgaag
421 gtccgcccca ctcaccgggg cgggccttcg cgttgagcta gagttcccgc cagcctcgca
481 gagcaggatt cccgttgagc accgccaggt gcgaataagg gacagtgaag aaggaacacc
541 cgctcgcggg tgggcctact tcacctatcc tgcccggctg acgccgttgg atacaccaag
601 gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg tgcgaaaaag gatggatata
661 ccgaaaaaat cgctataatg accccgaagc agggttatgc agcggaaaat gcagctcacg
721 gtaactgatg ccgtatttgc agtaccagcg tacggcccac agaatgatgt cacgctgaaa
781 atgccggcct ttgaatgggt tcatgtgcag ctccataagc aaaaggggat gataagttta
841 tcaccaccga ctatttgcaa cagtgccgtt gatcgtgcta tgatcgactg atgtcatcag
901 cggtggagtg caatgtcgtg caatacgaat ggcgaaaagc cgagctcatc ggtcagcttc
961 tcaaccttgg ggttaccccc ggcggtgtgc tgctggtcca cagctccttc cgtagcgtcc
1021 ggcccctcga agatgggcca cttggactga tcgaggccct gcgtgctgcg ctgggtccgg
1081 gagggacgct cgtcatgccc tcgtggtcag gtctggacga cgagccgttc gatcctgcca
1141 cgtcgcccgt tacaccggac cttggagttg tctctgactc attctggcgc ctgccaaatg
1201 taaagcgcag cgcccatcca tttgcctttg cggcagcggg gccacaggca gagcagatca
1261 tctctgatcc attgcccctg ccacctcact cgcctgcaag cccggtcgcc cgtgtccatg
1321 aactcgatgg gcaggtactt ctcctcggcg tgggacacga tgccaacacg acgctgcatc
1381 ttgccgagtt gttggcaaag gttccctatg gggtgccgag acactgcacc attcttcagg
1441 atggcaagtt ggtacgcgtc gattatctcg agaatgacca ctgctgtgag cgctttgcct
1501 tggcggacag gtggctcaag gagaagagcc ttcagaagga aggtccagtc ggtcatgcct
1561 ttgctcggtt gatccgctcc cgcgacattg tggcgacagc cctgggtcaa ctgggccgag
1621 atccgttgat cttcctgcat ccgccagagg cgggatgcga agaatgcgat gccgctcgcc
1681 agtcgattgg ctgagctcat aagttcctat tccgaagttc ctattcgaaa tgaccgacca
1741 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
1801 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
1861 gctggagttc ttcgcccacc ccaaaaggat ctaggtgaag atcctttttg ataatctcat
1921 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
1981 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
2041 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
2101 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
2161 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
2221 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
2281 gttaccggat aaggcgcagc ggtcgggctg aacggggggt cgtgcacaca gcccagcttg
2341 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
2401 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
2461 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
2521 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
2581 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
2641 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
2701 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
2761 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
2821 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
2881 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
2941 attgtgagcg gataacaatt tcacacagga aacagctatg acatgattac gaattgtacg
3001 cggtcgatct tgacggctgg cgagaggtgc ggggaggatc tgaccgacgc ggtccacacg
3061 tggcaccgcg atgctgttgt gggcacaatc gtgccggttg gtaggatcga cggccatggt
3121 tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg
3181 caccgagtcg gtgctttttt tacgtaggcg tcagctacga tgttccgggg actgctgatc
3241 cggtcagcag gtggaagagg gactggattc caaagttctc aatgctgctt gctgttcttg
3301 aatggggggt cgttgacgac gacatggctc gattggcgcg acaagttgct gcgattctca
3361 ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag
3421 gtcattactg gaccggatcg gggatctggg ctgagggagc cgacggcacg cggcggctca
3481 cggcgtggca cgcggaacgt ccgggcttgc acctcacgtc acgtgaggag gcagcgtgga
3541 cggcgtcaga gaagggagcg gacatatgtc ctccgagacc ggccccgtgg ccgtggaccc
3601 gaccctgcgt cgccgcatcg agccgcacga gttcgaggtc ttcttcgacc cgcgcgagct
3661 gcgcaaggag acctgcctgc tgtacgagat caactggggc ggccggcact ccatctggcg
3721 ccacacctcg cagaacacca acaagcacgt cgaggtcaac ttcatcgaga agttcaccac
3781 cgagcggtac ttctgcccga acacccgctg ctccatcacc tggttcctgt cctggtcccc
3841 ctgcggcgag tgctcccggg ccatcaccga gttcctgtcc cgctacccgc acgtcaccct
3901 gttcatctac atcgcccggc tgtaccacca cgccgacccg cgcaaccggc agggcctgcg
3961 cgacctgatc tcctccggcg tcaccatcca gatcatgacc gagcaggagt ccggctactg
4021 ctggcggaac ttcgtcaact actccccgtc caacgaggcc cactggcccc gctacccgca
4081 cctgtgggtc cgcctgtacg tcctggagct gtactgcatc atcctgggcc tgccgccctg
4141 cctgaacatc ctgcgtcgca agcagccgca gctgaccttc ttcaccatcg ccctccagtc
4201 ctgccactac cagcgcctgc cgccccacat cctgtgggcc accggcctga agtccggctc
4261 cgagaccccg ggcacctcgg agtccgccac cccggagtcc gacaagaagt actccatcgg
4321 cctcgcgatc ggcaccaact ccgtgggctg ggcggtcatc accgacgagt acaaggtccc
4381 ctccaagaag ttcaaggtcc tgggcaacac cgaccggcac tcgatcaaga agaacctgat
4441 cggcgccctg ctcttcgaca gcggcgagac cgccgaggcg acccgcctga agcggaccgc
4501 gcgtcgccgc tacacccggc gcaagaaccg catctgctac ctgcaggaaa tcttctccaa
4561 cgagatggcc aaggtggacg actcgttctt ccaccgcctg gaggagagct tcctggtgga
4621 ggaggacaag aagcacgagc gccacccgat cttcggcaac atcgtggacg aggtggccta
4681 ccacgagaag taccccacca tctaccacct ccgcaagaag ctggtggact cgaccgacaa
4741 ggcggacctg cggctcatct acctggccct cgcgcacatg atcaagttcc gcggccactt
4801 cctcatcgag ggcgacctga acccggacaa ctccgacgtg gacaagctct tcatccagct
4861 ggtgcagacc tacaaccagc tgttcgagga gaaccccatc aacgccagcg gcgtggacgc
4921 caaggcgatc ctctccgcgc gcctgagcaa gtcccggcgc ctggagaacc tcatcgccca
4981 gctgccgggc gagaagaaga acggcctctt cggcaacctg atcgcgctgt cgctcggcct
5041 gacccccaac ttcaagagca acttcgacct ggccgaggac gcgaagctcc agctgtccaa
5101 ggacacctac gacgacgacc tggacaacct gctcgcccag atcggcgacc agtacgcgga
5161 cctcttcctg gccgcgaaga acctctcgga cgccatcctg ctcagcgaca tcctgcgggt
5221 caacaccgag atcaccaagg ccccgctgtc ggcgagcatg atcaagcggt acgacgagca
5281 ccaccaggac ctgaccctgc tcaaggccct cgtgcgccag cagctgcccg agaagtacaa
5341 ggaaatcttc ttcgaccagt ccaagaacgg ctacgccggc tacatcgacg gcggcgcgtc
5401 gcaggaggag ttctacaagt tcatcaagcc gatcctggag aagatggacg gcaccgagga
5461 gctgctcgtc aagctgaacc gcgaggacct gctccgcaag cagcggacct tcgacaacgg
5521 ctccatcccg caccagatcc acctgggcga gctccacgcc atcctccggc gccaggagga
5581 cttctacccc ttcctgaagg acaaccgcga gaagatcgag aagatcctga ccttccgcat
5641 cccgtactac gtcggccccc tggcccgcgg caactcccgg ttcgcgtgga tgacccggaa
5701 gtcggaggag accatcaccc cgtggaactt cgaggaggtc gtggacaagg gcgcgtccgc
5761 gcagtcgttc atcgagcgca tgaccaactt cgacaagaac ctcccgaacg agaaggtcct
5821 gcccaagcac tccctgctct acgagtactt caccgtgtac aacgagctga ccaaggtcaa
5881 gtacgtgacc gagggcatgc ggaagccggc cttcctgtcg ggcgagcaga agaaggcgat
5941 cgtggacctg ctcttcaaga ccaaccgcaa ggtcaccgtg aagcagctga aggaggacta
6001 cttcaagaag atcgagtgct tcgactccgt cgagatcagc ggcgtggagg accgcttcaa
6061 cgcctccctg ggcacctacc acgacctgct caagatcatc aaggacaagg acttcctcga
6121 caacgaggag aacgaggaca tcctggagga catcgtcctc accctgaccc tcttcgagga
6181 ccgcgagatg atcgaggagc ggctcaagac ctacgcccac ctgttcgacg acaaggtgat
6241 gaagcagctg aagcgtcgcc gctacaccgg ctggggccgc ctctcccgga agctgatcaa
6301 cggcatccgg gacaagcaga gcggcaagac catcctggac ttcctcaagt ccgacggctt
6361 cgccaaccgc aacttcatgc agctcatcca cgacgacagc ctgaccttca aggaggacat
6421 ccagaaggcc caggtctcgg gccagggcga cagcctccac gagcacatcg ccaacctggc
6481 gggctccccg gcgatcaaga agggcatcct ccagaccgtc aaggtcgtgg acgagctggt
6541 caaggtgatg ggccgccaca agcccgagaa catcgtgatc gagatggccc gggagaacca
6601 gaccacccag aagggccaga agaactcgcg cgagcggatg aagcggatcg aggagggcat
6661 caaggagctc ggcagccaga tcctgaagga gcacccggtc gagaacaccc agctgcagaa
6721 cgagaagctg tacctctact acctgcagaa cggccgcgac atgtacgtgg accaggagct
6781 cgacatcaac cggctgtccg actacgacgt ggaccacatc gtgccgcagt ccttcctgaa
6841 ggacgactcg atcgacaaca aggtcctgac ccgctcggac aagaaccggg gcaagtccga
6901 caacgtgccc tcggaggagg tcgtgaagaa gatgaagaac tactggcgcc agctgctcaa
6961 cgccaagctc atcacccagc gcaagttcga caacctgacc aaggccgagc ggggcggcct
7021 gagcgagctc gacaaggcgg gcttcatcaa gcgccagctg gtcgagaccc ggcagatcac
7081 caagcacgtg gcccagatcc tggactcccg gatgaacacc aagtacgacg agaacgacaa
7141 gctgatccgc gaggtcaagg tgatcaccct caagagcaag ctggtctccg acttccgcaa
7201 ggacttccag ttctacaagg tccgggagat caacaactac caccacgccc acgacgcgta
7261 cctgaacgcc gtcgtgggca ccgcgctgat caagaagtac ccgaagctgg agtccgagtt
7321 cgtctacggc gactacaagg tctacgacgt gcgcaagatg atcgccaaga gcgagcagga
7381 gatcggcaag gccaccgcga agtacttctt ctactccaac atcatgaact tcttcaagac
7441 cgagatcacc ctggccaacg gcgagatccg caagcggccc ctgatcgaga ccaacggcga
7501 gaccggcgag atcgtctggg acaagggccg cgacttcgcc accgtccgga aggtgctgtc
7561 gatgccgcag gtcaacatcg tgaagaagac cgaggtgcag accggcggct tcagcaagga
7621 gtccatcctc cccaagcgca acagcgacaa gctgatcgcc cggaagaagg actgggaccc
7681 gaagaagtac ggcggcttcg acagccccac cgtcgcctac tccgtgctgg tcgtggcgaa
7741 ggtcgagaag ggcaagagca agaagctgaa gtccgtgaag gagctgctcg gcatcaccat
7801 catggagcgc tcctcgttcg agaagaaccc gatcgacttc ctggaggcca agggctacaa
7861 ggaggtcaag aaggacctca tcatcaagct gcccaagtac agcctgttcg agctggagaa
7921 cggccgcaag cggatgctcg cctccgcggg cgagctgcag aagggcaacg agctggccct
7981 cccgtcgaag tacgtcaact tcctgtacct cgcgtcccac tacgagaagc tgaagggctc
8041 gcccgaggac aacgagcaga agcagctctt cgtggagcag cacaagcact acctggacga
8101 gatcatcgag cagatcagcg agttcagcaa gcgcgtcatc ctggccgacg cgaacctcga
8161 caaggtgctg tccgcctaca acaagcaccg cgacaagccg atccgggagc aggcggagaa
8221 catcatccac ctgttcaccc tcaccaacct gggcgccccc gccgcgttca agtacttcga
8281 caccaccatc gaccgcaagc ggtacacctc caccaaggag gtcctcgacg cgaccctgat
8341 ccaccagagc atcaccggcc tgtacgagac ccgcatcgac ctgtcccagc tcggcggcga
8401 ctccggcggc tccaccaacc tgtccgacat catcgagaag gagaccggca agcagctggt
8461 catccaggag tccatcctga tgctgccgga ggaagtcgag gaagtcatcg gcaacaagcc
8521 cgagtccgac atcctggtcc acaccgccta cgacgagtcc accgacgaga acgtcatgct
8581 gctgacctcg gacgcccccg agtacaagcc ctgggccctg gtcatccagg actccaacgg
8641 cgagaacaag atcaagatgc tgtgatctag aggatccccg ggtaccgagc tcgaattccc
8701 cagatctaaa gttttgtcgt ctttccagac gttagtaaat gaattttctg tatgaggttt
8761 tgctaaacaa ctttcaacag tttcagcgga gtgagaatag aaaggaacaa ctaaaggaat
8821 tgcgaataat aattttttca cgttgaaaat ctccaaaaaa aaaggctcca aaaggagcct
8881 ttaattgtat cggtttatca gcttgctttc gaggtgaatt tcttaaacag cttgataccg
8941 atagttgcgc cgacaatgac aacaaccatc gcccacgcat aaccgatata ttcggtcgct
9001 gaggcttgca gggagtcaaa ggccgctttt gcgggatctc gtcgaaggca ctagaggcct
9061 atatgcgggg atcgaccgcg cgggtcccgg acggggaaga gcggggagct ttgccagaga
9121 gcgacgactt ccccttgcgt tggtgattgc cggtcagggc agccatccgc catcgtcgcg
9181 tagggtgtca caccccagga atcgcgtcac tgaacacagc agccggtagg acgaccatga
9241 ctgagttgga caccatcgca aatccgtccg atcccgcggt gcagcggatc atcgatgtca
9301 ccaagccgtc gcgatccaac ataaagacaa cgttgatcga ggacgtcgag cccctcatgc
9361 acagcatcgc ggccggggtg gagttcatcg aggtctacgg cagcgacagc agtccttttc
9421 catctgagtt gctggatctg tgcgggcggc agaacatacc ggtccgcctc atcgactcct
9481 cgatcgtcaa ccagttgttc aagggggagc ggaaggccaa gacattcggc atcgcccgcg
9541 tccctcgccc ggccaggttc ggcgatatcg cgagccggcg tggggacgtc gtcgttctcg
9601 acggggtgaa gatcgtcggg aacatcggcg cgatagtacg cacgtcgctc gcgctcggag
9661 cgtcggggat catcctggtc gacagtgaca tcaccagcat cgcggaccgg cgtctccaaa
9721 gggccagccg aggttacgtc ttctcccttc ccgtcgttct ctccggtcgc gaggaggcca
9781 tcgccttcat tcgggacagc ggtatgcagc tgatgacgct caaggcggat ggcgacattt
9841 ccgtgaagga actcggggac aatccggatc ggctggcctt gctgttcggc agcgaaaagg
9901 gtgggccttc cgacctgttc gaggaggcgt cttccgcctc ggtttccatc cccatgatga
9961 gccagaccga gtctctcaac gtttccgttt ccctcggaat cgcgctgcac gagaggatcg
10021 acaggaatct cgcggccaac cgataagcgc ctctgttcct cggacgctcg gttgctcgac
10081 ctcgattcgt cagtgatgat caacaggacc cagaaatggc acgagcccgg gattggcctc
10141 ccgggctcgc cgcccccgaa ccggcgcagc atgccgagtg aggacgtttg ctctgtgacg
10201 agcgtacgag agaacagcgc gtcgacacca ccgcgccccg ccggaccgaa gtgccactgc
10261 ggcaccccga tcgtccacat ccccgggaag cgccccaaaa tctactgctc ggacgcctgc
10321 cgaaagcgcg caaagcgtct cattgcggac cagtcacgca gcgtagcaag cacgactaca
10381 gcgggtaatc cggacgcggg ctttagtagt aagggaaaat ctccgtccac tgcaggtcag
10441 agcgctgtaa ctggaacggg ttctacccca aacgagggtg ccgcgacagc cgggcgggag
10501 gacccgcgcg acgagcggtt cgcccggcgg gaccggcacc agacggtgtc gctgaacgag
10561 gcgttcaagg gctgcgggac ccggctcact gccgggacgg ccgagctgat gtggaagccg
10621 ggcgaggcga cgtggggcaa cacgtgccgc tgcaacaacg tccacacctg cccgtggtgc
10681 atgagccgca tcctggccgt acggggctcg aacgtgcagc tcgccgccga cggtctcgcg
10741 gacgccgggt acgggctgca ccttggcacg aacacgttgc gccacttcga gcgcatggcg
10801 ttcggcaccg tccgcaaggg aatgcggcac ggcctggtcg cggtcttgca cgacggctgg
10861 aagggcgcgt acggctcgtc cggccgccgc tggcgcacga tgcgcgacga cttcgggatc
10921 atcggctacg agcgggcctt cgaggacacc ttcggctggg gctccggctg gcacctgcac
10981 tggcacacgc tctgggtcac ccgcgaggtg ctcggcccgg acgcccaggc cgcgttccgc
11041 gacgcactcg ccggagcctg ggccgccggg gtcgaatccg cgggcggcta caccgtcagc
11101 gagacgtgcg accggcccgg ttgctcctgt gagggcaagg gccacggcac cgacgtgcgc
11161 ccactcaacg gcgcggacgc cgctgacggc gatgcgggca agcaggcccg ctacctctac
11221 aaggacggcg acaagaccaa gggcggcgtc gccaagatcg gcctggagct ggccggacag
11281 aacttcaagg ccggtcgcgg cgacgaccgc atgggaccgc tcgacctggg cgacgcggcg
11341 gccgccgagc tgcagcggct tcgtcggccc ggcccgttcg tcgagaagta ccgcgagcgc
11401 gagttcggcg tcttccaggt ccgcaagcac taccggtcgc agaacctgaa ccggctgatc
11461 aaggaactcg gcatccagca ggacgtccgc accgaggagg agatcaccga cgacaccgag
11521 ggcctggtcg ccatcgcggt catcccggcc tacatctggt accgatacat cgcccgcgtc
11581 gccggacgtc gcctcgacct catcaaggtc gccgagacgt acggcctacc gggcgtccgg
11641 cggctcgtcg agtcctgggg cctggtgtgg ggcaaggacg tcctggaccc gcccgccccg
11701 gaagcgccgg ccgcgccggg cgaccttgac gctgaccaga tgaggttcga ggtcatgtcc
11761 gaggaggaag ctgctttccg cgaggcgcgc cggaaggcca acgaggcccg cacggaggag
11821 ctggccgctt cgctcgaccg agtgcgtcag ccgaagaagg aggcgatccg cccgacgatc
11881 agccttcgga agcgcctgaa gccgaagccg gtcaccgtcg acgtcaagac tcccccgccc
11941 ggcgccgcgt cgcccgtgtg ccgccgctgc aagggcaagc tcgccccggt actccagccc
12001 tggggccagc accccggaga ctgcctccgc gtcgacacag cggtcgcgta gggccgctga
12061 gggaccgcca tgagaccgcc accccggaca tccgggtggc ggtctcatgg cggtcccggc
12121 gcgaaagccg agggtctgcc tgccgtgagg tggccggcgg acctgggtta ctgctggaat
12181 cccgcgccca gtccgagcgt ccgccggcgc ttttcacctg gctccgggtg ctcgtcgatg
12241 agcacca
//
