| 质粒名称: | pEMPTY::sgRNA2 |
|---|---|
| 目录编号: | 165459 |
| 存储实验室: | Inigo Lasa |
|
载体骨架基本信息 |
|
| 载体名称: | pCN38, pMAD |
| 完整质粒大小: | 11277 bp |
| 载体类型: | Bacterial Expression, CRISPR, Synthetic Biology |
| 筛选标记: | In Gram-positive bacteria: chloramphenicol |
| 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: | gRNA / shRNA |
| 物种来源: | Staphylococcus aureus |
| 启动子: | SP01 |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | AscI(未破坏) |
| 3’克隆位点: | SmaI (未破坏) |
| 5’测序引物 | cgatttttgtgatgctcgtcaggg |
| 3’测序引物 | cgtaaacggatgctggctag |
|
文献引用方法 |
|
| 材料和方法部分: | pEMPTY::sgRNA2 was a gift from Inigo Lasa (Addgene plasmid # 165459 ; http://n2t.net/addgene:165459 ; RRID:Addgene_165459) |
| 参考文献部分: | Fitness Cost Evolution of Natural Plasmids of Staphylococcus aureus. Dorado-Morales P, Garcillan-Barcia MP, Lasa I, Solano C. mBio. 2021 Feb 23;12(1). pii: mBio.03094-20. doi: 10.1128/mBio.03094-20. 10.1128/mBio.03094-20 PubMed 33622733 |
买家导航
pEMPTY::sgRNA2载体质粒基本信息
pEMPTY::sgRNA2质粒图谱载体图谱和pEMPTY::sgRNA2载体序列质粒序列多克隆位点信息
pEMPTY::sgRNA2质粒载体简介
pEMPTY::sgRNA2质粒序列载体序列
LOCUS pEMPTY::sgRNA2 11278 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Escherichia coli – Staphylococcus aureus shuttle vector for plasmid
curing in Gram-positive bacteria.
ACCESSION .
VERSION .
KEYWORDS pEMPTY::sgRNA2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11278)
AUTHORS Dorado-Morales P, Garcillan-Barcia MP, Lasa I, Solano C
TITLE Fitness Cost Evolution of Natural Plasmids of Staphylococcus aureus.
JOURNAL mBio. 2021 Feb 23;12(1). pii: mBio.03094-20. doi:
10.1128/mBio.03094-20.
PUBMED 33622733
REFERENCE 2 (bases 1 to 11278)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..11278
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 1176..1198
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(1236..1254)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 1322..1426
/gene="bla"
/label=AmpR promoter
CDS 1427..2287
/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 complement(1645..1664)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2458..3046
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2947..2966
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
misc_RNA 3260..3335
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
terminator 3416..3463
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind 3459..3477
/label=T7 Term
/note="T7 terminator, reverse primer"
CDS complement(3512..7618)
/codon_start=1
/product="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
/label=Cas9
/note="generates RNA-guided double strand breaks in DNA"
/translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
protein_bind 7661..7679
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O1 for the tetR and tetA genes"
CDS 7789..8412
/codon_start=1
/gene="tetR from transposon Tn10"
/product="tetracycline repressor TetR"
/label=TetR
/note="TetR binds to the tetracycline operator tetO to
inhibit transcription. This inhibition can be relieved by
adding tetracycline or doxycycline."
/translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
primer_bind complement(7862..7881)
/label=Tet-R
/note="Tetracycline resistance gene, reverse primer"
primer_bind complement(8493..8510)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(8493..8509)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8502..8524)
/label=M13/pUC Forward
/note="In lacZ gene"
ORIGIN
1 ataggcttgg ttatgccggt actgccgggc ctcttgcggg atcgaggggg cccacctagg
61 tattatcaag ataagaaaga aaaggatttt tcgctacgct caaatccttt aaaaaaacac
121 aaaagaccac attttttaat gtggtctttt attcttcaac taaagcaccc attagttcaa
181 caaacgaaaa ttggataaag tgggatattt ttaaaatata tatttatgtt acagtaatat
241 tgacttttaa aaaaggattg attctaatga agaaagcaga caagtaagcc tcctaaattc
301 actttagata aaaatttagg aggcatatca aatgaacttt aataaaattg atttagacaa
361 ttggaagaga aaagagatat ttaatcatta tttgaaccaa caaacgactt ttagtataac
421 cacagaaatt gatattagtg ttttataccg aaacataaaa caagaaggat ataaatttta
481 ccctgcattt attttcttag tgacaagggt gataaactca aatacagctt ttagaactgg
541 ttacaatagc gacggagagt taggttattg ggataagtta gagccacttt atacaatttt
601 tgatggtgta tctaaaacat tctctggtat ttggactcct gtaaagaatg acttcaaaga
661 gttttatgat ttataccttt ctgatgtaga gaaatataat ggttcgggga aattgtttcc
721 caaaacacct atacctgaaa atgctttttc tctttctatt attccatgga cttcatttac
781 tgggtttaac ttaaatatca ataataatag taattacctt ctacccatta ttacagcagg
841 aaaattcatt aataaaggta attcaatata tttaccgcta tctttacagg tacatcattc
901 tgtttgtgat ggttatcatg caggattgtt tatgaactct attcaggaat tgtcagatag
961 gcctaatgac tggcttttat aatatgagat aatgccgact gtacttttta cagtcggttt
1021 tctaatgtca ctaacctgcc ccgttagttg aagaaggccg cggcctcgag cggccgcata
1081 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
1141 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
1201 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
1261 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
1321 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
1381 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
1441 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
1501 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
1561 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
1621 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
1681 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
1741 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
1801 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
1861 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
1921 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
1981 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
2041 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
2101 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
2161 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
2221 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
2281 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
2341 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
2401 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
2461 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
2521 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
2581 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
2641 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
2701 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
2761 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
2821 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
2881 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
2941 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
3001 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
3061 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
3121 atcccctgat tctgtggata accgtattac cgcctttgag tgagctggcg gccgctgcat
3181 gcctgcaggg cgcgccttga caaattgcag taggcatgac aaaatggact cacaagtttt
3241 gggattgtta agggttccgg ttttagagct agaaatagca agttaaaata aggctagtcc
3301 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttcgcaaaaa accccgcttc
3361 ggcggggttt tttcgctagt cggtgtgcgc aaccttaagc ccgggtacca gatctcaaaa
3421 aacccctcaa gacccgttta gaggccccaa ggggttatgc tagttattgc tcagcggtgg
3481 cagcagccta ggataccggt aacatgggat ctcagtcacc tcctagctga ctcaaatcaa
3541 tgcgtgtttc ataaagacca gtgatggatt gatggataag agtggcatct aaaacttctt
3601 ttgtagacgt atatcgttta cgatcaattg ttgtatcaaa atatttaaaa gcagcgggag
3661 ctccaagatt cgtcaacgta aataaatgaa taatattttc tgcttgttca cgtattggtt
3721 tgtctctatg tttgttatat gcactaagaa ctttatctaa attggcatct gctaaaataa
3781 cacgcttaga aaattcactg atttgctcaa taatctcatc taaataatgc ttatgctgct
3841 ccacaaacaa ttgtttttgt tcgttatctt ctggactacc cttcaacttt tcataatgac
3901 tagctaaata taaaaaattc acatatttgc ttggcagagc cagctcattt cctttttgta
3961 attctccggc actagccagc atccgtttac gaccgttttc taactcaaaa agactatatt
4021 taggtagttt aatgattaag tcttttttaa cttccttata tcctttagct tctaaaaagt
4081 caatcggatt tttttcaaag gaacttcttt ccataattgt gatccctagt aactctttaa
4141 cggattttaa cttcttcgat ttcccttttt ccaccttagc aaccactagg actgaataag
4201 ctaccgttgg actatcaaaa ccaccatatt tttttggatc ccagtctttt ttacgagcaa
4261 taagcttgtc cgaatttctt tttggtaaaa ttgactcctt ggagaatccg cctgtctgta
4321 cttctgtttt cttgacaata ttgacttggg gcatggacaa tactttgcgc actgtggcaa
4381 aatctcgccc tttatcccag acaatttctc cagtttcccc attagtttcg attagagggc
4441 gtttgcgaat ctctccattt gcaagtgtaa tttctgtttt gaagaagttc atgatattag
4501 agtaaaagaa atattttgcg gttgctttgc ctatttcttg ctcagactta gcaatcattt
4561 tacgaacatc ataaacttta taatcaccat agacaaactc cgattcaagt tttggatatt
4621 tcttaatcaa agcagttcca acgacggcat ttagatacgc atcatgggca tgatggtaat
4681 tgttaatctc acgtacttta tagaattgga aatcttttcg gaagtcagaa actaatttag
4741 attttaaggt aatcacttta acctctcgaa taagtttatc attttcatcg tatttagtat
4801 tcatgcgact atccaaaatt tgtgccacat gcttagtgat ttggcgagtt tcaaccaatt
4861 ggcgtttgat aaaaccagct ttatcaagtt cactcaaacc tccacgttca gctttcgtta
4921 aattatcaaa cttacgttga gtgattaact tggcgtttag aagttgtctc caatagtttt
4981 tcatcttttt gactacttct tcacttggaa cgttatccga tttaccacga tttttatcag
5041 aacgcgttaa gaccttattg tctattgaat cgtctttaag gaaactttgt ggaacaatgt
5101 gatcgacatc ataatcactt aaacgattaa tatctaattc ttggtccaca tacatgtctc
5161 ttccattttg gagataatag agatagagct tttcattttg caattgagta ttttcaacag
5221 gatgctcttt aagaatctga cttcctaatt ctttgatacc ttcttcgatt cgtttcatac
5281 gctctcgcga atttttctgg cccttttgag ttgtctgatt ttcacgtgcc atttcaataa
5341 cgatattttc tggcttatgc cgccccatta ctttgaccaa ttcatcaaca acttttacag
5401 tctgtaaaat acctttttta atagcagggc taccagctaa atttgcaata tgttcatgta
5461 aactatcgcc ttgtccagac acttgtgctt tttgaatgtc ttctttaaat gtcaaactat
5521 catcatggat cagctgcata aaattgcgat tggcaaaacc atctgatttc aaaaaatcta
5581 atattgtttt gccagattgc ttatccctaa taccattaat caattttcga gacaaacgtc
5641 cccaaccagt ataacggcga cgtttaagct gtttcatcac cttatcatca aagaggtgag
5701 catatgtttt aagtctttcc tcaatcatct ccctatcttc aaataaggtc aatgttaaaa
5761 caatatcctc taagatatct tcattttctt cattatccaa aaaatcttta tctttaataa
5821 tttttagcaa atcatggtag gtacctaatg aagcattaaa tctatcttca actcctgaaa
5881 tttcaacact atcaaaacat tctatttttt tgaaataatc ttcttttaat tgcttaacgg
5941 ttacttttcg atttgttttg aagagtaaat caacaatggc tttcttctgt tcacctgaaa
6001 gaaatgctgg ttttcgcatt ccttcagtaa catatttgac ctttgtcaat tcgttataaa
6061 ccgtaaaata ctcataaagc aaactatgtt ttggtagtac tttttcattt ggaagatttt
6121 tatcaaagtt tgtcatgcgt tcaataaatg attgagctga agcaccttta tcgacaactt
6181 cttcaaaatt ccatggggta attgtttctt cagacttccg agtcatccat gcaaaacgac
6241 tattgccacg cgccaatgga ccaacataat aaggaattcg aaaagtcaag attttttcaa
6301 tcttctcacg attgtctttt aaaaatggat aaaagtcttc ttgtcttctc aaaatagcat
6361 gcagctcacc caagtgaatt tgatggggaa tagagccgtt gtcaaaggtc cgttgcttgc
6421 gcagcaaatc ttcacgattt agtttcacca ataattcctc agtaccatcc attttttcta
6481 aaattggttt gataaattta taaaattctt cttggctagc tcccccatca atataacctg
6541 catatccgtt ttttgattga tcaaaaaaga tttctttata cttttctgga agttgttgtc
6601 gaactaaagc ttttaaaaga gtcaagtctt gatgatgttc atcgtagcgt ttaatcattg
6661 aagctgatag gggagcctta gttatttcag tatttactct taggatatct gaaagtaaaa
6721 tagcatctga taaattctta gctgccaaaa acaaatcagc atattgatct ccaatttgcg
6781 ccaataaatt atctaaatca tcatcgtaag tatcttttga aagctgtaat ttagcatctt
6841 ctgccaaatc aaaatttgat ttaaaattag gggtcaaacc caatgacaaa gcaatgagat
6901 tcccaaataa gccatttttc ttctcaccgg ggagctgagc aatgagattt tctaatcgtc
6961 ttgatttact caatcgtgca gaaagaatcg ctttagcatc tactccactt gcgttaatag
7021 ggttttcttc aaataattga ttgtaggttt gtaccaactg gataaatagt ttgtccacat
7081 cactattatc aggatttaaa tctccctcaa tcaaaaaatg accacgaaac ttaatcatat
7141 gcgctaaggc caaatagatt aagcgcaaat ccgctttatc agtagaatct accaattttt
7201 ttcgcagatg atagatagtt ggatatttct catgataagc aacttcatct actatatttc
7261 caaaaatagg atgacgttca tgcttcttgt cttcttccac caaaaaagac tcttcaagtc
7321 gatgaaagaa actatcatct actttcgcca tctcatttga aaaaatctcc tgtagataac
7381 aaatacgatt cttccgacgt gtataccttc tacgagctgt ccgtttgaga cgagtcgctt
7441 ccgctgtctc tccactgtca aataaaagag cccctataag attttttttg atactgtggc
7501 ggtctgtatt tcccagaacc ttgaactttt tagacggaac cttatattca tcagtgatca
7561 ccgcccatcc gacgctattt gtgccgatat ctaagcctat tgagtatttc ttatccataa
7621 ataatcatcc tcctaaagat ctcaagctta ttttaattat actctatcaa tgatagagtg
7681 tcaatatttt ttttagtttt tcatgaactc gaggggatcc aaataaaaaa ctagtttgac
7741 aaataactct atcaatgata taatgtcaac aaaaaggagg aattaatgat gtctagatta
7801 gataaaagta aagtgattaa cagcgcatta gagctgctta atgaggtcgg aatcgaaggt
7861 ttaacaaccc gtaaactcgc ccagaagcta ggtgtagagc agcctacatt gtattggcat
7921 gtaaaaaata agcgggcttt gctcgacgcc ttagccattg agatgttaga taggcaccat
7981 actcactttt gccctttaga aggggaaagc tggcaagatt ttttacgtaa taacgctaaa
8041 agttttagat gtgctttact aagtcatcgc gatggagcaa aagtacattt aggtacacgg
8101 cctacagaaa aacagtatga aactctcgaa aatcaattag cctttttatg ccaacaaggt
8161 ttttcactag agaatgcatt atatgcactc agcgctgtgg ggcattttac tttaggttgc
8221 gtattggaag atcaagagca tcaagtcgct aaagaagaaa gggaaacacc tactactgat
8281 agtatgccgc cattattacg acaagctatc gaattatttg atcaccaagg tgcagagcca
8341 gccttcttat tcggccttga attgatcata tgcggattag aaaaacaact taaatgtgaa
8401 agtgggtctt aaaagcagca taaccttttt ccgtgatggt aacttcacgg taaccaagat
8461 gtcgagttat cgataccgtc gacctggaat tcactggccg tcgttttaca acgtcgtgac
8521 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
8581 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
8641 ggcgaatggc gcctgagggt tgccagagtt aaaggatcaa gtaatacaaa cgggatacaa
8701 agacataatc aaagagagaa taaaaactat aataataaag acataaatca tgaggaaaca
8761 tataaaaatt atgatttgat taacgcacaa aatataaagt ataaagataa aattgatgaa
8821 acgattgatg agaattattc agggaaacgt aaaattcggt cagatgcaat tcgacatgtg
8881 gacggactgg ttacaagtga taaagatttc tttgatgatt taagcggaga agaaatagaa
8941 cgatttttta aagatagctt ggagtttcta gaaaatgaat acggtaagga aaatatgctg
9001 tatgcgactg tccatctgga tgaaagagtc ccacatatgc actttggttt tgtcccttta
9061 acagaggacg ggagattgtc tgcaaaagaa cagttaggca acaagaaaga ctttactcaa
9121 ttacaagata gatttaatga gtatgtgaat gagaaaggtt atgaacttga aagaggcacg
9181 tccaaagagg ttacagaacg agaacataaa gcgatggatc agtacaagaa agatactgta
9241 tttcataaac aggaactgca agaagttaag gatgagttac agaaggcaaa taagcagtta
9301 cagagtggaa tagagcatat gaggtctacg aaaccctttg attatgaaaa tgagcgtaca
9361 ggtttgttct ctggacgtga agagactggt agaaagatat taactgctga tgaatttgaa
9421 cgcctgcaag aaacaatctc ttctgcagaa cggattgttg atgattacga aaatattaag
9481 agcacagact attacacaga aaatcaagaa ttaaaaaaac gtagagagag tttgaaagaa
9541 gtagtgaata catggaaaga ggggtatcac gaaaaaagta aagaggttaa taaattaaag
9601 cgagagaatg atagtttgaa tgagcagttg aatgtatcag agaaatttca agatagtaca
9661 gtgactttat atcgtgctgc gagggcgaat ttccctgggt ttgagaaagg gtttaatagg
9721 cttaaagaga aattctttaa tgattccaaa ttcgagcgtg tgggacagtt tatggatgtt
9781 gtacaggata atgtccagaa ggtcgataga aagcgtgaga aacagcgtac agacgattta
9841 gagatgtaga ggtactttta tgccgagaaa actttttgcg tgtgacagtc cttaaaatat
9901 acttagagcg taagcgaaag tagtagcgac agctattaac tttcggttgc aaagctctag
9961 gatttttaat ggacgcagcg catcacacgc aaaaaggaaa ttggaataaa tgcgaaattt
10021 gagatgttaa ttaaagacct ttttgaggtc tttttttctt agatttttgg ggttatttag
10081 gggagaaaac ataggggggt actacgacct cccccctagg tgtccattgt ccattgtcca
10141 aacaaataaa taaatattgg gtttttaatg ttaaaaggtt gttttttatg ttaaagtgaa
10201 aaaaacagat gttgggaggt acagtgatgg ttgtagatag aaaagaagag aaaaaagttg
10261 ctgttacttt aagacttaca acagaagaaa atgagatatt aaatagaatc aaagaaaaat
10321 ataatattag caaatcagat gcaaccggta ttctaataaa aaaatatgca aaggaggaat
10381 acggtgcatt ttaaacaaaa aaagatagac agcactggca tgctgcctat ctatgactaa
10441 attttgttaa atgtattagc accgttatta tatcatgagc gaaaatgtaa taaaagaaac
10501 tgaaaacaag aaaaattcaa gaggacgtaa ttggacattt gttttatatc cagaatcagc
10561 aaaagccgag tggttagagt atttaaaaga gttacacatt caatttgtag tgtctccatt
10621 acatgatagg gatactgata cagaagatag gatgaaaaaa gagcattatc atattctagt
10681 gatgtatgag ggtaataaat cttatgaaca gataaaaata attacagaag aattgaatgc
10741 gactattccg cagattgcag gaagtgtgaa aggtcttgtg agatatatgc ttcacatgga
10801 cgatcctaat aaatttaaat atcaaaaaga agatatgata gtttatggcg gtgtagatgt
10861 tgatgaatta ttaaagaaaa caacaacaga tagatataaa ttaattaaag aaatgattga
10921 gtttattgat gaacaaggaa tcgtagaatt taagagttta atggattatg caatgaagtt
10981 taaatttgat gattggttcc cgcttttatg tgataactcg gcgtatgtta ttcaagaata
11041 tataaaatca aatcggtata aatctgaccg atagattttg aatttaagag tgtcacaaga
11101 cactcttttt tcgcaccaac gaaaactggt ttaagccgac tgcgcaaaag acataatcga
11161 taagctttaa tgcggtagtt tatcacagtt aaattgctaa cgcagtcagg caccgtgtat
11221 gaaatctaac aatgcgctca tcgtcatcct cggcaccgtc accctggatg ctgtaggc
//
