pV1200

111429 Synthetic Yeast Expression

Plasmid Information

Purpose
CaCas9/gRNA Solo entry plasmid for cloning guides - contains stuffer with BsmBI sites - integrates at ENO1 - Recyclable for serial mutagenesis (Sap2-FLP)
Depositor
Gerald Fink
Article
Vyas et al Sci Adv. 2015;1(3):e1500248.
Insert
CaCas9/sgRNA-BsmBI stuffer
Available Since
Oct. 17, 2018
Availability
Available

Sequence Information

Length
16,324 bp
Topology
circular
Molecule Type
other DNA

Features

CDS 7
Label Gene Product Position Strand
Cas9 Cas9 (Csn1) endonuclease from the Strep… 2244..6348 Fwd
DNA Sequence
ATGGATAAAAAGTATAGTATTGGTTTAGATATTGGTACTAACTCTGTGGGTTGGGCAGTTATCACTGACGAATATAAAGTTCCATCAAAGAAATTTAAGGTGTTAGGTAACACTGACAGACACTCAATAAAAAAGAATCTTATCGGTGCTCTTTTGTTCGACTCCGGTGAAACTGCCGAGGCTACACGTTTAAAAAGAACAGCAAGAAGAAGATATACCCGTAGAAAAAATAGAATATGTTATTTACAAGAAATCTTTTCTAATGAAATGGCTAAAGTTGATGATTCCTTTTTCCATAGATTGGAAGAGTCATTTTTGGTTGAAGAAGACAAAAAGCATGAGAGACATCCAATCTTTGGGAATATAGTTGATGAAGTGGCTTACCATGAAAAATATCCTACCATTTATCATTTAAGAAAGAAATTGGTAGATTCAACTGATAAAGCTGACCTTAGATTAATCTATTTAGCACTTGCCCATATGATTAAATTTAGAGGTCATTTTTTGATTGAAGGTGATTTGAACCCAGATAATTCTGACGTGGATAAATTATTTATTCAATTAGTCCAAACCTACAACCAATTATTTGAGGAAAATCCAATTAATGCTAGTGGTGTCGATGCCAAAGCTATATTATCAGCCAGATTATCAAAATCTAGACGTTTGGAAAATTTGATTGCCCAATTGCCAGGAGAAAAAAAGAATGGATTATTTGGAAACTTGATCGCATTATCATTGGGTTTGACACCAAATTTTAAATCTAATTTTGATTTAGCTGAAGATGCTAAATTACAATTATCAAAAGACACCTATGACGACGATTTGGACAATTTACTTGCTCAAATTGGTGATCAATATGCAGATTTGTTCTTAGCTGCTAAAAACTTATCTGATGCTATTTTGTTGTCTGATATTTTGAGAGTGAACACAGAAATAACCAAAGCTCCATTATCAGCATCTATGATCAAACGTTATGATGAACACCATCAGGATTTGACTTTATTGAAAGCTTTGGTGAGACAACAATTGCCAGAGAAGTATAAAGAAATCTTTTTCGATCAATCTAAAAACGGGTATGCAGGTTATATTGATGGGGGTGCCTCCCAAGAGGAATTTTACAAATTTATAAAACCTATTTTAGAAAAGATGGATGGGACTGAGGAACTTTTGGTCAAATTGAACAGAGAAGATTTGTTACGTAAACAGAGAACTTTTGATAATGGTAGTATACCTCACCAAATTCATTTGGGTGAGTTGCATGCAATTTTAAGAAGACAAGAAGATTTTTATCCATTTTTAAAAGATAATAGAGAAAAAATCGAGAAAATTTTAACCTTTAGAATTCCATACTATGTTGGGCCTTTGGCTAGAGGTAATTCAAGATTTGCCTGGATGACACGTAAATCAGAAGAAACTATTACCCCTTGGAATTTTGAAGAGGTTGTTGATAAAGGAGCATCAGCACAGAGTTTTATTGAAAGAATGACCAATTTCGATAAAAACTTACCAAATGAAAAAGTTTTACCAAAACATTCCTTGTTATACGAATATTTTACTGTTTACAATGAACTTACAAAGGTTAAATATGTTACTGAAGGTATGCGTAAGCCAGCCTTTTTATCTGGAGAACAGAAAAAGGCAATAGTTGATTTATTGTTTAAAACAAATAGAAAAGTTACTGTTAAACAATTAAAAGAAGATTACTTTAAGAAAATTGAATGTTTTGATTCAGTTGAAATCAGTGGTGTTGAAGACAGATTTAATGCTAGTTTAGGAACTTACCATGATTTACTTAAAATTATCAAAGATAAAGATTTCTTGGATAACGAAGAAAATGAAGACATTTTAGAAGACATTGTTTTAACCTTAACTTTATTCGAAGATAGAGAGATGATTGAAGAACGTTTGAAGACTTATGCACATTTGTTTGACGATAAAGTGATGAAACAGTTGAAAAGAAGACGTTATACTGGATGGGGTAGATTGTCTCGTAAATTGATCAATGGAATTAGAGATAAACAAAGTGGTAAAACTATCTTGGACTTTTTGAAATCTGACGGATTTGCTAATAGAAATTTCATGCAATTGATCCACGACGATAGTTTGACATTTAAAGAAGACATCCAAAAGGCCCAAGTGAGTGGGCAAGGTGATTCATTACATGAACATATTGCAAATTTAGCCGGATCTCCTGCTATTAAGAAAGGGATATTACAAACTGTTAAAGTTGTGGATGAATTAGTGAAAGTAATGGGAAGACATAAACCTGAAAACATTGTCATTGAGATGGCAAGAGAAAATCAAACTACACAAAAAGGACAGAAAAATAGTAGAGAACGTATGAAAAGAATAGAAGAGGGTATTAAAGAATTGGGTAGTCAAATATTGAAAGAACACCCAGTGGAAAATACCCAGTTGCAAAATGAAAAATTATATCTTTACTACCTTCAAAATGGACGTGATATGTATGTTGATCAGGAATTAGATATAAATAGACTTTCAGATTATGATGTAGATCATATAGTTCCACAATCTTTCTTGAAAGATGATTCCATAGACAATAAAGTATTAACTAGAAGTGATAAAAATAGAGGTAAAAGTGATAATGTCCCAAGTGAGGAAGTCGTCAAAAAGATGAAAAATTACTGGCGTCAACTTTTGAATGCTAAATTAATTACTCAAAGAAAATTTGATAATTTGACTAAAGCAGAAAGAGGTGGGCTTTCTGAATTAGATAAAGCCGGGTTCATTAAAAGACAATTGGTCGAAACTAGACAAATTACTAAACATGTTGCCCAAATTTTAGATTCCCGTATGAACACTAAGTATGACGAAAATGATAAGTTAATACGTGAGGTTAAAGTCATTACTTTAAAATCAAAACTTGTCTCTGATTTCAGAAAGGATTTCCAATTCTATAAAGTTAGAGAAATTAATAATTATCATCATGCTCATGATGCATATTTGAATGCTGTAGTTGGAACTGCTTTAATCAAGAAATACCCTAAATTAGAATCTGAATTTGTATATGGTGATTACAAAGTCTATGATGTTAGAAAGATGATTGCTAAATCAGAACAAGAAATTGGTAAAGCTACAGCTAAATACTTCTTTTACTCTAACATTATGAATTTCTTTAAAACAGAAATTACTTTGGCAAACGGTGAAATTAGAAAAAGACCTCTTATTGAAACAAATGGTGAGACTGGAGAGATAGTTTGGGACAAAGGGCGTGATTTCGCTACTGTTAGAAAAGTTTTATCAATGCCACAAGTTAACATTGTAAAGAAAACAGAGGTTCAAACTGGTGGTTTCTCAAAAGAAAGTATTTTGCCTAAAAGAAATAGTGATAAATTGATTGCCAGAAAAAAGGATTGGGATCCAAAGAAATATGGTGGTTTCGACTCACCAACCGTAGCCTATTCTGTTTTGGTTGTGGCAAAGGTTGAAAAGGGTAAAAGTAAAAAGCTTAAATCAGTAAAAGAACTTTTGGGTATTACAATAATGGAAAGAAGTTCCTTTGAAAAGAACCCTATTGATTTTTTGGAAGCTAAAGGTTATAAGGAAGTAAAGAAGGACTTAATAATCAAATTGCCTAAATATTCTTTATTTGAATTAGAAAATGGGAGAAAAAGAATGTTGGCTTCTGCTGGAGAATTGCAAAAGGGTAATGAATTAGCATTGCCTTCCAAATATGTTAACTTCTTGTATTTAGCTTCACACTATGAAAAGTTGAAAGGGTCACCAGAAGATAACGAGCAAAAACAATTATTTGTTGAACAACACAAACACTACTTAGATGAGATTATAGAACAAATTAGTGAATTCAGTAAAAGAGTGATATTAGCTGATGCAAATTTAGATAAAGTTTTGTCAGCCTATAACAAACATAGAGATAAGCCAATTAGAGAACAAGCAGAAAACATTATTCACTTATTTACCCTTACCAATTTAGGAGCACCTGCTGCTTTCAAGTATTTTGATACAACAATTGATCGTAAAAGATATACCTCAACAAAAGAAGTCTTAGACGCCACCTTAATTCATCAATCAATCACTGGATTGTATGAGACAAGAATTGATTTGTCTCAATTGGGTGGTGAT
Protein Sequence
MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD
SV40 NLS nuclear localization signal of SV40 (si… 6360..6381 Fwd
DNA Sequence
CCTAAGAAGAAAAGAAAAGTT
Protein Sequence
PKKKRKV
SV40 NLS nuclear localization signal of SV40 (si… 6384..6405 Fwd
DNA Sequence
CCAAAGAAAAAGCGTAAGGTG
Protein Sequence
PKKKRKV
SV40 NLS nuclear localization signal of SV40 (si… 6408..6429 Fwd
DNA Sequence
CCTAAGAAAAAGAGAAAGGTT
Protein Sequence
PKKKRKV
3xFLAG three tandem FLAG(R) epitope tags, foll… 6429..6495 Fwd
DNA Sequence
GACTACAAAGACCATGACGGTGATTATAAAGATCATGACATCGACTACAAGGATGACGATGACAAG
Protein Sequence
DYKDHDGDYKDHDIDYKDDDDK
FLP site-specific recombinase 7956..9228 Fwd
DNA Sequence
ATGCCACAATTTGATATATTATGTAAAACACCACCTAAGGTGCTTGTTCGTCAGTTTGTGGAAAGGTTTGAAAGACCTTCAGGTGAGAAAATAGCATTATGTGCTGCTGAACTAACCTATTTATGTTGGATGATTACACATAACGGAACAGCAATCAAGAGAGCCACATTCATGAGCTATAATACTATCATAAGCAATTCGTTGAGTTTCGATATTGTCAATAAATCACTCCAGTTTAAATACAAGACGCAAAAAGCAACAATTTTGGAAGCCTCATTAAAGAAATTGATTCCTGCTTGGGAATTTACAATTATTCCTTACTATGGACAAAAACATCAATCTGATATCACTGATATTGTAAGTAGTTTGCAATTACAGTTCGAATCATCGGAAGAAGCAGATAAGGGGAATAGCCACAGTAAAAAAATGCTTAAAGCACTTCTAAGTGAGGGTGAAAGCATCTGGGAGATCACTGAGAAAATACTAAATTCGTTTGAGTATACTTCGAGATTTACAAAAACAAAAACTTTATACCAATTCCTCTTCCTAGCTACTTTCATCAACTGTGGAAGATTCAGCGATATTAAGAACGTTGATCCGAAATCATTTAAATTAGTCCAAAATAAGTATTTGGGAGTAATAATCCAGTGTTTAGTGACAGAGACAAAGACAAGCGTTAGTAGGCACATATACTTCTTTAGCGCAAGGGGTAGGATCGATCCACTTGTATATTTGGATGAATTTTTGAGGAATTCTGAACCAGTCCTAAAACGAGTAAATAGGACCGGCAATTCTTCAAGCAATAAACAGGAATACCAATTATTAAAAGATAACTTAGTCAGATCGTACAATAAAGCTTTGAAGAAAAATGCGCCTTATTCAATCTTTGCTATAAAAAATGGCCCAAAATCTCACATTGGAAGACATTTGATGACCTCATTTCTTTCAATGAAGGGCCTAACGGAGTTGACTAATGTTGTGGGAAATTGGAGCGATAAGCGTGCTTCTGCCGTGGCCAGGACAACGTATACTCATCAGATAACAGCAATACCTGATCACTACTTCGCACTAGTTTCTCGGTACTATGCATATGATCCAATATCAAAGGAAATGATAGCATTGAAGGATGAGACTAATCCAATTGAGGAGTGGCAGCATATAGAACAGCTAAAGGGTAGTGCTGAAGGAAGCATACGATACCCCGCATGGAATGGGATAATATCACAGGAGGTACTAGACTACCTTTCATCCTACATAAATAGACGCATATAA
Protein Sequence
MPQFDILCKTPPKVLVRQFVERFERPSGEKIALCAAELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEFTIIPYYGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLKDNLVRSYNKALKKNAPYSIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI
AmpR bla beta-lactamase 15350..16211 Rev
DNA Sequence
ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA
Protein Sequence
MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
promoter 5
Label Gene Product Position Strand
T7 promoter 640..659 Fwd
DNA Sequence
TAATACGACTCACTATAGG
TEF promoter 10452..10796 Rev
DNA Sequence
GACATGGAGGCCCAGAATACCCTCCTTGACAGTCTTGACGTGCGCAGCTCAGGGGCATGATGTGACTGTCGCCCGTACATTTAGCCCATACATCCCCATGTATAATCATTTGCATCCATACATTTTGATGGCCGCACGGCGCGAAGCAAAAATTACGGCTCCTCGCTGCAGACCTGCGAGCAGGGAAACGCTCCCCTCACAGACGCGTTGAATTGTCCCCACGCCGCGCCCCTGTAGAGAAATATAAAAGGTTAGGATTTGCCACTGAGGTTCTTCTTTCATATACTTCCTTTTAAAATCTTGCTAGGATACAGTTCTCACATCACATCCGAACATAAACAACC
T3 promoter 14150..14169 Rev
DNA Sequence
AATTAACCCTCACTAAAGG
lac promoter 14237..14268 Rev
DNA Sequence
TTTACACTTTATGCTCCCGGCTCGTATGTTG
AmpR promoter bla 16211..16316 Rev
DNA Sequence
CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
protein_bind 5
Label Gene Product Position Strand
FRT (minimal) 6826..6860 Rev
DNA Sequence
GAAGTTCCTATTCTCTAGAAAGTATAGGAACTTC
FRT (minimal) 12018..12052 Fwd
DNA Sequence
GAAGTTCCTATTCTCTAGAAAGTATAGGAACTTC
FRT (minimal) 12058..12092 Rev
DNA Sequence
GAAGTTCCTATTCTCTAGAAAGTATAGGAACTTC
lac operator 14213..14230 Fwd
DNA Sequence
TTGTTATCCGCTCACAA
CAP binding site 14282..14304 Fwd
DNA Sequence
ATGAGTGAGCTAACTCACATTA
rep_origin 2
Label Gene Product Position Strand
f1 ori 17..473 Rev
DNA Sequence
ACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGCTTACAATTT
ori 14591..15180 Rev
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA
terminator 2
Label Gene Product Position Strand
CYC1 terminator S. cerevisiae CYC1 6559..6807 Fwd
DNA Sequence
TCATGTAATTAGTTATGTCACGCTTACATTCACGCCCTCCCCCCACATCCGCTCTAACCGAAAAGGAAGGAGTTAGACAACCTGAAGTCTAGGTCCCTATTTATTTTTTTATAGTTATGTTAGTATTAAGAACGTTATTTATATTTCAAATTTTTCTTTTTTTTCTGTACAGACGCGTGTACGCATGTAACATTATACTGAAAACCTTGCTTGAGAAGGTTTTGGGACGCTCGAAGGCTTTAATTTGC
TEF terminator 9654..9852 Rev
DNA Sequence
ACTGACAATAAAAAGATTCTTGTTTTCAAGAACTTGTCATTTGTATAGTTTTTTTATATTGTAGTTGTTCTATTTTAATCAAATGTTAGCGTGATTTATATTTTTTTTCGCCTCGACATCATCTGCCCAGATGCGAAGTTAAGTGCGCAGAAAGTAATATCATGCGTCAATCGTATGTGAATGCTGGTCGCTATACTG