| 质粒名称: | pCRISPR-aBEST |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 131464 |
| 存储实验室: | Tilmann Weber |
|
载体骨架基本信息 |
|
| 载体名称: | pGM1190 |
| 空载体大小: | 6971 bp |
| 完整质粒大小: | 12452 bp |
| 载体类型: | CRISPR,合成生物学 |
| 载体抗性: | 安普霉素 |
| 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: | Streptomyces codon optimized spCas9n (D10A) |
| 物种来源: | Synthetic; Streptomyces |
| 插入大小: | 4104 bp |
| GenBank编号 | KR011748 |
| 启动子: | tipA |
| 克隆方法: | Gibson克隆 |
| 5’测序引物 | gcggcagcagcggcggctcggacaagaagtactccatcgg |
| 3’测序引物 | ggggatcctctagaaagctttcagtcgccgccgagctggg |
|
文献引用方法 |
|
| 材料和方法部分: | pCRISPR-aBEST was a gift from Tilmann Weber (Addgene plasmid # 131464 ; http://n2t.net/addgene:131464 ; RRID:Addgene_131464) |
| 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
pCRISPRaBEST载体基本信息
pCRISPRaBEST载体图谱质粒图谱和多克隆位点信息
pCRISPRaBEST载体简介
pCRISPRaBEST载体序列
LOCUS pCRISPR-aBEST 12449 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expression of a Streptomyces codon optimized Cas9n-tadA fusion
protein in pGM1190. A to G base editor for actinomycetes.
ACCESSION .
VERSION .
KEYWORDS pCRISPR-aBEST
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12449)
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 12449)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..12449
/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 3120..3195
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
terminator 3365..3399
/label=lambda t0 terminator
/note="minimal transcription terminator from phage lambda
(Scholtissek and Grosse, 1987)"
CDS 4758..8861
/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"
terminator 9048..9096
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
CDS 10811..12253
/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 cggccatggg
3121 ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg
3181 gcaccgagtc ggtgcttttt ttacgtaggc gtcagctacg atgttccggg gactgctgat
3241 ccggtcagca ggtggaagag ggactggatt ccaaagttct caatgctgct tgctgttctt
3301 gaatgggggg tcgttgacga cgacatggct cgattggcgc gacaagttgc tgcgattctc
3361 accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga
3421 ggtcattact ggaccggatc ggggatctgg gctgagggag ccgacggcac gcggcggctc
3481 acggcgtggc acgcggaacg tccgggcttg cacctcacgt cacgtgagga ggcagcgtgg
3541 acggcgtcag agaagggagc ggacatatga gcgaggtcga gttctcccac gagtactgga
3601 tgcggcacgc cctgaccctg gcgaagcggg cctgggacga gcgcgaggtc cccgtcggcg
3661 ccgtcctggt ccacaacaac cgggtcatcg gcgagggctg gaaccggccc atcggccggc
3721 acgaccccac cgcccacgcc gagatcatgg ccctgcgcca gggcggcctg gtcatgcaga
3781 actaccggct gatcgacgcc accctgtacg tcaccctgga gccctgcgtc atgtgcgccg
3841 gcgccatgat ccactcccgc atcggccggg tcgtcttcgg cgcccgcgac gccaagaccg
3901 gcgccgccgg ctccctgatg gacgtcctgc accaccccgg catgaaccac cgcgtcgaga
3961 tcaccgaggg catcctggcc gacgagtgcg ccgccctgct gtccgacttc ttccggatgc
4021 gccggcagga gatcaaggcc cagaagaagg cccagtcctc caccgactcc ggcggctcct
4081 ccggcggctc ctccggctcc gagacgccgg gcacctcgga gtccgccacc ccggagtcgt
4141 ccggcggctc ctccggcggc tcctccgagg tcgagttctc gcacgagtac tggatgcgcc
4201 acgccctgac cctggccaag cgggcccggg acgaacgcga ggtcccggtg ggcgcggtcc
4261 tggtcctgaa caaccgggtg attggtgagg gctggaaccg cgccatcggc ctgcacgacc
4321 cgaccgcgca cgcggagatc atggcgctcc ggcagggcgg cctggtgatg cagaactacc
4381 gcctcatcga cgcgaccctc tacgtcacct tcgagccctg cgtgatgtgc gcgggcgcga
4441 tgattcactc ccgcattggc cgggtggtct tcggcgtccg caacgcgaag accggcgcgg
4501 cgggctccct gatggacgtg ctgcactacc cgggcatgaa ccaccgggtg gaaattaccg
4561 agggcatcct ggcggacgag tgcgcggcgc tgctgtgcta cttcttccgc atgccccggc
4621 aggtcttcaa cgcgcagaag aaggcccaat cctccaccga ttccggcggc tcctccggcg
4681 gctcctccgg ctccgaaacc ccgggcacct cggagtcggc caccccggag tccagcggcg
4741 gcagcagcgg cggctcggac aagaagtact ccatcggcct cgcgatcggc accaactccg
4801 tgggctgggc ggtcatcacc gacgagtaca aggtcccctc caagaagttc aaggtcctgg
4861 gcaacaccga ccggcactcg atcaagaaga acctgatcgg cgccctgctc ttcgacagcg
4921 gcgagaccgc cgaggcgacc cgcctgaagc ggaccgcgcg tcgccgctac acccggcgca
4981 agaaccgcat ctgctacctg caggaaatct tctccaacga gatggccaag gtggacgact
5041 cgttcttcca ccgcctggag gagagcttcc tggtggagga ggacaagaag cacgagcgcc
5101 acccgatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct
5161 accacctccg caagaagctg gtggactcga ccgacaaggc ggacctgcgg ctcatctacc
5221 tggccctcgc gcacatgatc aagttccgcg gccacttcct catcgagggc gacctgaacc
5281 cggacaactc cgacgtggac aagctcttca tccagctggt gcagacctac aaccagctgt
5341 tcgaggagaa ccccatcaac gccagcggcg tggacgccaa ggcgatcctc tccgcgcgcc
5401 tgagcaagtc ccggcgcctg gagaacctca tcgcccagct gccgggcgag aagaagaacg
5461 gcctcttcgg caacctgatc gcgctgtcgc tcggcctgac ccccaacttc aagagcaact
5521 tcgacctggc cgaggacgcg aagctccagc tgtccaagga cacctacgac gacgacctgg
5581 acaacctgct cgcccagatc ggcgaccagt acgcggacct cttcctggcc gcgaagaacc
5641 tctcggacgc catcctgctc agcgacatcc tgcgggtcaa caccgagatc accaaggccc
5701 cgctgtcggc gagcatgatc aagcggtacg acgagcacca ccaggacctg accctgctca
5761 aggccctcgt gcgccagcag ctgcccgaga agtacaagga aatcttcttc gaccagtcca
5821 agaacggcta cgccggctac atcgacggcg gcgcgtcgca ggaggagttc tacaagttca
5881 tcaagccgat cctggagaag atggacggca ccgaggagct gctcgtcaag ctgaaccgcg
5941 aggacctgct ccgcaagcag cggaccttcg acaacggctc catcccgcac cagatccacc
6001 tgggcgagct ccacgccatc ctccggcgcc aggaggactt ctaccccttc ctgaaggaca
6061 accgcgagaa gatcgagaag atcctgacct tccgcatccc gtactacgtc ggccccctgg
6121 cccgcggcaa ctcccggttc gcgtggatga cccggaagtc ggaggagacc atcaccccgt
6181 ggaacttcga ggaggtcgtg gacaagggcg cgtccgcgca gtcgttcatc gagcgcatga
6241 ccaacttcga caagaacctc ccgaacgaga aggtcctgcc caagcactcc ctgctctacg
6301 agtacttcac cgtgtacaac gagctgacca aggtcaagta cgtgaccgag ggcatgcgga
6361 agccggcctt cctgtcgggc gagcagaaga aggcgatcgt ggacctgctc ttcaagacca
6421 accgcaaggt caccgtgaag cagctgaagg aggactactt caagaagatc gagtgcttcg
6481 actccgtcga gatcagcggc gtggaggacc gcttcaacgc ctccctgggc acctaccacg
6541 acctgctcaa gatcatcaag gacaaggact tcctcgacaa cgaggagaac gaggacatcc
6601 tggaggacat cgtcctcacc ctgaccctct tcgaggaccg cgagatgatc gaggagcggc
6661 tcaagaccta cgcccacctg ttcgacgaca aggtgatgaa gcagctgaag cgtcgccgct
6721 acaccggctg gggccgcctc tcccggaagc tgatcaacgg catccgggac aagcagagcg
6781 gcaagaccat cctggacttc ctcaagtccg acggcttcgc caaccgcaac ttcatgcagc
6841 tcatccacga cgacagcctg accttcaagg aggacatcca gaaggcccag gtctcgggcc
6901 agggcgacag cctccacgag cacatcgcca acctggcggg ctccccggcg atcaagaagg
6961 gcatcctcca gaccgtcaag gtcgtggacg agctggtcaa ggtgatgggc cgccacaagc
7021 ccgagaacat cgtgatcgag atggcccggg agaaccagac cacccagaag ggccagaaga
7081 actcgcgcga gcggatgaag cggatcgagg agggcatcaa ggagctcggc agccagatcc
7141 tgaaggagca cccggtcgag aacacccagc tgcagaacga gaagctgtac ctctactacc
7201 tgcagaacgg ccgcgacatg tacgtggacc aggagctcga catcaaccgg ctgtccgact
7261 acgacgtgga ccacatcgtg ccgcagtcct tcctgaagga cgactcgatc gacaacaagg
7321 tcctgacccg ctcggacaag aaccggggca agtccgacaa cgtgccctcg gaggaggtcg
7381 tgaagaagat gaagaactac tggcgccagc tgctcaacgc caagctcatc acccagcgca
7441 agttcgacaa cctgaccaag gccgagcggg gcggcctgag cgagctcgac aaggcgggct
7501 tcatcaagcg ccagctggtc gagacccggc agatcaccaa gcacgtggcc cagatcctgg
7561 actcccggat gaacaccaag tacgacgaga acgacaagct gatccgcgag gtcaaggtga
7621 tcaccctcaa gagcaagctg gtctccgact tccgcaagga cttccagttc tacaaggtcc
7681 gggagatcaa caactaccac cacgcccacg acgcgtacct gaacgccgtc gtgggcaccg
7741 cgctgatcaa gaagtacccg aagctggagt ccgagttcgt ctacggcgac tacaaggtct
7801 acgacgtgcg caagatgatc gccaagagcg agcaggagat cggcaaggcc accgcgaagt
7861 acttcttcta ctccaacatc atgaacttct tcaagaccga gatcaccctg gccaacggcg
7921 agatccgcaa gcggcccctg atcgagacca acggcgagac cggcgagatc gtctgggaca
7981 agggccgcga cttcgccacc gtccggaagg tgctgtcgat gccgcaggtc aacatcgtga
8041 agaagaccga ggtgcagacc ggcggcttca gcaaggagtc catcctcccc aagcgcaaca
8101 gcgacaagct gatcgcccgg aagaaggact gggacccgaa gaagtacggc ggcttcgaca
8161 gccccaccgt cgcctactcc gtgctggtcg tggcgaaggt cgagaagggc aagagcaaga
8221 agctgaagtc cgtgaaggag ctgctcggca tcaccatcat ggagcgctcc tcgttcgaga
8281 agaacccgat cgacttcctg gaggccaagg gctacaagga ggtcaagaag gacctcatca
8341 tcaagctgcc caagtacagc ctgttcgagc tggagaacgg ccgcaagcgg atgctcgcct
8401 ccgcgggcga gctgcagaag ggcaacgagc tggccctccc gtcgaagtac gtcaacttcc
8461 tgtacctcgc gtcccactac gagaagctga agggctcgcc cgaggacaac gagcagaagc
8521 agctcttcgt ggagcagcac aagcactacc tggacgagat catcgagcag atcagcgagt
8581 tcagcaagcg cgtcatcctg gccgacgcga acctcgacaa ggtgctgtcc gcctacaaca
8641 agcaccgcga caagccgatc cgggagcagg cggagaacat catccacctg ttcaccctca
8701 ccaacctggg cgcccccgcc gcgttcaagt acttcgacac caccatcgac cgcaagcggt
8761 acacctccac caaggaggtc ctcgacgcga ccctgatcca ccagagcatc accggcctgt
8821 acgagacccg catcgacctg tcccagctcg gcggcgactg aaagctttct agaggatccc
8881 cgggtaccga gctcgaattc cccagatcta aagttttgtc gtctttccag acgttagtaa
8941 atgaattttc tgtatgaggt tttgctaaac aactttcaac agtttcagcg gagtgagaat
9001 agaaaggaac aactaaagga attgcgaata ataatttttt cacgttgaaa atctccaaaa
9061 aaaaaggctc caaaaggagc ctttaattgt atcggtttat cagcttgctt tcgaggtgaa
9121 tttcttaaac agcttgatac cgatagttgc gccgacaatg acaacaacca tcgcccacgc
9181 ataaccgata tattcggtcg ctgaggcttg cagggagtca aaggccgctt ttgcgggatc
9241 tcgtcgaagg cactagaggc ctatatgcgg ggatcgaccg cgcgggtccc ggacggggaa
9301 gagcggggag ctttgccaga gagcgacgac ttccccttgc gttggtgatt gccggtcagg
9361 gcagccatcc gccatcgtcg cgtagggtgt cacaccccag gaatcgcgtc actgaacaca
9421 gcagccggta ggacgaccat gactgagttg gacaccatcg caaatccgtc cgatcccgcg
9481 gtgcagcgga tcatcgatgt caccaagccg tcgcgatcca acataaagac aacgttgatc
9541 gaggacgtcg agcccctcat gcacagcatc gcggccgggg tggagttcat cgaggtctac
9601 ggcagcgaca gcagtccttt tccatctgag ttgctggatc tgtgcgggcg gcagaacata
9661 ccggtccgcc tcatcgactc ctcgatcgtc aaccagttgt tcaaggggga gcggaaggcc
9721 aagacattcg gcatcgcccg cgtccctcgc ccggccaggt tcggcgatat cgcgagccgg
9781 cgtggggacg tcgtcgttct cgacggggtg aagatcgtcg ggaacatcgg cgcgatagta
9841 cgcacgtcgc tcgcgctcgg agcgtcgggg atcatcctgg tcgacagtga catcaccagc
9901 atcgcggacc ggcgtctcca aagggccagc cgaggttacg tcttctccct tcccgtcgtt
9961 ctctccggtc gcgaggaggc catcgccttc attcgggaca gcggtatgca gctgatgacg
10021 ctcaaggcgg atggcgacat ttccgtgaag gaactcgggg acaatccgga tcggctggcc
10081 ttgctgttcg gcagcgaaaa gggtgggcct tccgacctgt tcgaggaggc gtcttccgcc
10141 tcggtttcca tccccatgat gagccagacc gagtctctca acgtttccgt ttccctcgga
10201 atcgcgctgc acgagaggat cgacaggaat ctcgcggcca accgataagc gcctctgttc
10261 ctcggacgct cggttgctcg acctcgattc gtcagtgatg atcaacagga cccagaaatg
10321 gcacgagccc gggattggcc tcccgggctc gccgcccccg aaccggcgca gcatgccgag
10381 tgaggacgtt tgctctgtga cgagcgtacg agagaacagc gcgtcgacac caccgcgccc
10441 cgccggaccg aagtgccact gcggcacccc gatcgtccac atccccggga agcgccccaa
10501 aatctactgc tcggacgcct gccgaaagcg cgcaaagcgt ctcattgcgg accagtcacg
10561 cagcgtagca agcacgacta cagcgggtaa tccggacgcg ggctttagta gtaagggaaa
10621 atctccgtcc actgcaggtc agagcgctgt aactggaacg ggttctaccc caaacgaggg
10681 tgccgcgaca gccgggcggg aggacccgcg cgacgagcgg ttcgcccggc gggaccggca
10741 ccagacggtg tcgctgaacg aggcgttcaa gggctgcggg acccggctca ctgccgggac
10801 ggccgagctg atgtggaagc cgggcgaggc gacgtggggc aacacgtgcc gctgcaacaa
10861 cgtccacacc tgcccgtggt gcatgagccg catcctggcc gtacggggct cgaacgtgca
10921 gctcgccgcc gacggtctcg cggacgccgg gtacgggctg caccttggca cgaacacgtt
10981 gcgccacttc gagcgcatgg cgttcggcac cgtccgcaag ggaatgcggc acggcctggt
11041 cgcggtcttg cacgacggct ggaagggcgc gtacggctcg tccggccgcc gctggcgcac
11101 gatgcgcgac gacttcggga tcatcggcta cgagcgggcc ttcgaggaca ccttcggctg
11161 gggctccggc tggcacctgc actggcacac gctctgggtc acccgcgagg tgctcggccc
11221 ggacgcccag gccgcgttcc gcgacgcact cgccggagcc tgggccgccg gggtcgaatc
11281 cgcgggcggc tacaccgtca gcgagacgtg cgaccggccc ggttgctcct gtgagggcaa
11341 gggccacggc accgacgtgc gcccactcaa cggcgcggac gccgctgacg gcgatgcggg
11401 caagcaggcc cgctacctct acaaggacgg cgacaagacc aagggcggcg tcgccaagat
11461 cggcctggag ctggccggac agaacttcaa ggccggtcgc ggcgacgacc gcatgggacc
11521 gctcgacctg ggcgacgcgg cggccgccga gctgcagcgg cttcgtcggc ccggcccgtt
11581 cgtcgagaag taccgcgagc gcgagttcgg cgtcttccag gtccgcaagc actaccggtc
11641 gcagaacctg aaccggctga tcaaggaact cggcatccag caggacgtcc gcaccgagga
11701 ggagatcacc gacgacaccg agggcctggt cgccatcgcg gtcatcccgg cctacatctg
11761 gtaccgatac atcgcccgcg tcgccggacg tcgcctcgac ctcatcaagg tcgccgagac
11821 gtacggccta ccgggcgtcc ggcggctcgt cgagtcctgg ggcctggtgt ggggcaagga
11881 cgtcctggac ccgcccgccc cggaagcgcc ggccgcgccg ggcgaccttg acgctgacca
11941 gatgaggttc gaggtcatgt ccgaggagga agctgctttc cgcgaggcgc gccggaaggc
12001 caacgaggcc cgcacggagg agctggccgc ttcgctcgac cgagtgcgtc agccgaagaa
12061 ggaggcgatc cgcccgacga tcagccttcg gaagcgcctg aagccgaagc cggtcaccgt
12121 cgacgtcaag actcccccgc ccggcgccgc gtcgcccgtg tgccgccgct gcaagggcaa
12181 gctcgccccg gtactccagc cctggggcca gcaccccgga gactgcctcc gcgtcgacac
12241 agcggtcgcg tagggccgct gagggaccgc catgagaccg ccaccccgga catccgggtg
12301 gcggtctcat ggcggtcccg gcgcgaaagc cgagggtctg cctgccgtga ggtggccggc
12361 ggacctgggt tactgctgga atcccgcgcc cagtccgagc gtccgccggc gcttttcacc
12421 tggctccggg tgctcgtcga tgagcacca
//
