pCashGem-Xyl2-URA3-LINEAR

174840 Synthetic Bacterial Expression

Plasmid Information

Purpose
pRS414 backbone containing LINEAR fragment targeting XYL2 in S. stipitis
Depositor
Zengyi Shao
Article
Ploessl et al Nat Chem Biol. 2021 Oct 28. pii: 10.1038
Insert
Codon optimized Cas9:hGem
Tags
hGeminin tag
Promoter
ENO1p
Available Since
Jan. 5, 2022
Availability
Available

Sequence Information

Length
15,413 bp
Topology
circular
Molecule Type
other DNA
View on AddGene.org

Features

CDS 8
Label Gene Product Position Strand
AmpR bla beta-lactamase 759..1620 Rev
DNA Sequence
ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA
Protein Sequence
MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
TRP1 S. cerevisiae TRP1 phosphoribosylanthranilate isomerase, r… 2804..3479 Fwd
DNA Sequence
ATGTCTGTTATTAATTTCACAGGTAGTTCTGGTCCATTGGTGAAAGTTTGCGGCTTGCAGAGCACAGAGGCCGCAGAATGTGCTCTAGATTCCGATGCTGACTTGCTGGGTATTATATGTGTGCCCAATAGAAAGAGAACAATTGACCCGGTTATTGCAAGGAAAATTTCAAGTCTTGTAAAAGCATATAAAAATAGTTCAGGCACTCCGAAATACTTGGTTGGCGTGTTTCGTAATCAACCTAAGGAGGATGTTTTGGCTCTGGTCAATGATTACGGCATTGATATCGTCCAACTGCATGGAGATGAGTCGTGGCAAGAATACCAAGAGTTCCTCGGTTTGCCAGTTATTAAAAGACTCGTATTTCCAAAAGACTGCAACATACTACTCAGTGCAGCTTCACAGAAACCTCATTCGTTTATTCCCTTGTTTGATTCAGAAGCAGGTGGGACAGGTGAACTTTTGGATTGGAACTCGATTTCTGACTGGGTTGGAAGGCAAGAGAGCCCCGAAAGCTTACATTTTATGTTAGCTGGTGGACTGACGCCAGAAAATGTTGGTGATGCGCTTAGATTAAATGGCGTTATTGGTGTTGATGTAAGCGGAGGTGTGGAGACAAATGGTGTAAAAGACTCTAACAAAATAGCAAATTTCGTCAAAAATGCTAAGAAATAG
Protein Sequence
MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRKRTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDESWQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDWVGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNAKK
FLAG — FLAG(R) epitope tag, followed by an ent… 5681..5705 Fwd
DNA Sequence
GACTACAAGGATGACGATGACAAG
Protein Sequence
DYKDDDDK
SV40 NLS — nuclear localization signal of SV40 (si… 5708..5729 Fwd
DNA Sequence
CCAAAGAAAAAGAGAAAGGTT
Protein Sequence
PKKKRKV
Cas9 — Cas9 (Csn1) endonuclease from the Strep… 5729..9830 Fwd
DNA Sequence
GACAAGAAGTACTCTATCGGTTTGGACATCGGCACTAACTCTGTTGGTTGGGCTGTTATCACTGACGAGTACAAGGTCCCATCCAAGAAGTTCAAGGTCTTGGGTAACACCGACAGACACTCCATCAAGAAGAACTTGATCGGCGCCTTGTTGTTCGACTCTGGTGAAACTGCTGAAGCCACCAGATTGAAGAGAACCGCCAGAAGAAGATACACCAGAAGAAAGAACAGAATCTGCTACTTGCAAGAGATCTTCTCCAACGAAATGGCCAAGGTCGACGACTCGTTCTTCCACAGATTGGAAGAATCCTTCTTGGTCGAAGAGGACAAGAAGCACGAAAGACACCCAATCTTCGGTAACATCGTTGACGAAGTCGCCTACCACGAAAAGTACCCAACCATCTACCACTTGAGAAAGAAGTTGGTCGACTCTACCGACAAGGCCGACTTGAGATTGATCTACTTGGCTTTGGCCCACATGATCAAGTTCAGAGGTCACTTCTTGATCGAGGGCGACTTGAACCCAGATAACTCTGACGTTGACAAGTTGTTCATCCAGTTGGTCCAGACCTACAACCAGTTGTTCGAAGAAAACCCAATCAACGCTTCTGGTGTCGACGCTAAGGCTATCTTGTCTGCCAGATTGTCCAAGTCCAGAAGATTGGAAAACTTGATCGCCCAGTTGCCAGGCGAAAAGAAGAACGGTTTGTTCGGCAACTTGATCGCTTTGTCCTTGGGCTTGACCCCCAACTTCAAGTCTAACTTCGACTTGGCTGAGGACGCCAAGTTGCAGTTGTCTAAGGATACTTACGACGACGACTTGGACAACTTGTTGGCTCAAATCGGTGACCAGTACGCTGACTTGTTCTTGGCTGCTAAGAACTTGTCTGACGCCATCTTGTTGTCCGACATCTTGAGAGTCAACACCGAAATCACCAAGGCCCCATTGTCTGCCTCTATGATCAAGAGATACGACGAACACCACCAGGACTTGACCTTGTTGAAGGCTTTGGTTAGACAGCAGTTGCCCGAGAAGTACAAAGAAATCTTCTTCGACCAGTCCAAGAACGGTTACGCTGGTTACATCGATGGTGGTGCCTCCCAAGAAGAGTTCTACAAGTTCATCAAGCCCATCTTGGAAAAGATGGACGGCACCGAAGAGTTGTTGGTCAAGTTGAACAGAGAGGACTTGTTGAGAAAGCAGAGAACCTTCGACAACGGTTCTATCCCACATCAGATCCACTTGGGTGAATTGCACGCCATCTTGAGAAGACAAGAGGACTTCTACCCATTCTTGAAGGACAACAGAGAAAAGATCGAGAAGATCTTGACCTTCAGAATCCCCTACTACGTCGGTCCATTGGCCAGAGGTAATTCTAGATTCGCCTGGATGACCAGAAAGTCCGAAGAAACTATCACCCCCTGGAACTTCGAAGAAGTTGTTGATAAGGGCGCCTCCGCCCAGTCTTTCATCGAAAGAATGACCAACTTCGACAAGAACTTGCCCAACGAGAAGGTCTTGCCAAAGCACTCCTTGTTGTACGAGTACTTCACCGTCTACAACGAGTTGACCAAGGTCAAGTACGTCACCGAAGGTATGAGAAAGCCAGCCTTCTTGTCTGGTGAACAGAAGAAGGCTATCGTCGACTTGTTGTTCAAGACCAACAGAAAGGTCACCGTCAAGCAGTTGAAAGAGGACTACTTCAAGAAAATCGAGTGCTTCGACTCCGTCGAAATCTCTGGTGTCGAAGATAGATTCAACGCCTCCTTGGGTACTTACCACGACTTGTTGAAGATCATCAAGGACAAGGATTTCTTGGACAACGAAGAGAACGAGGACATCTTGGAGGACATCGTCTTGACTTTGACCTTGTTCGAGGACAGAGAGATGATCGAAGAGAGATTGAAAACCTACGCCCACTTGTTCGACGACAAGGTCATGAAGCAGTTGAAGAGAAGAAGATATACCGGCTGGGGCAGATTGTCCAGAAAGTTGATCAACGGTATCAGAGACAAGCAGTCCGGCAAGACTATCTTGGACTTCTTGAAGTCTGACGGCTTCGCCAACAGAAACTTCATGCAGTTGATCCACGACGATTCCTTGACCTTCAAAGAGGACATCCAGAAGGCTCAAGTTTCTGGTCAAGGTGACTCCTTGCACGAACATATCGCTAACTTGGCTGGTTCTCCAGCCATCAAGAAGGGTATCTTGCAGACTGTTAAGGTCGTCGACGAATTGGTGAAGGTCATGGGTAGACACAAGCCAGAGAACATCGTCATCGAAATGGCTAGAGAAAACCAGACCACCCAGAAGGGTCAGAAGAACTCCAGAGAAAGAATGAAGAGAATCGAAGAGGGCATCAAAGAGTTGGGCTCCCAGATCTTGAAAGAACACCCAGTCGAAAACACCCAGTTGCAGAACGAGAAGTTGTACTTGTACTACTTGCAGAACGGCAGAGACATGTACGTCGACCAAGAGTTGGACATCAACAGATTGTCTGACTACGACGTTGACCACATCGTCCCCCAATCTTTCTTGAAGGATGACTCCATCGACAACAAGGTCTTGACCAGATCCGACAAGAACAGAGGTAAGTCTGACAACGTTCCATCCGAAGAGGTCGTCAAGAAGATGAAGAACTATTGGAGACAGTTGTTGAACGCCAAGTTGATCACCCAGAGAAAGTTCGACAACTTGACTAAGGCCGAAAGAGGTGGTTTGTCTGAATTGGACAAGGCCGGCTTCATCAAGAGACAGTTGGTCGAAACTAGACAGATCACCAAGCACGTCGCTCAGATCTTGGACTCCAGAATGAACACCAAGTACGACGAGAACGACAAGTTGATTAGAGAGGTCAAGGTCATCACCTTGAAGTCCAAGTTGGTGTCCGACTTCAGAAAGGACTTCCAGTTCTACAAGGTCAGAGAGATCAACAACTACCACCACGCTCATGACGCTTACTTGAACGCTGTTGTCGGTACTGCCTTGATCAAGAAGTACCCAAAGTTGGAATCCGAGTTCGTCTACGGTGACTACAAGGTCTACGACGTCAGAAAGATGATCGCCAAGTCCGAACAAGAAATCGGTAAGGCTACCGCCAAGTACTTCTTCTACTCCAACATCATGAATTTCTTCAAGACCGAGATCACCTTGGCCAACGGCGAGATCAGAAAAAGACCATTGATCGAAACTAACGGCGAAACCGGTGAAATCGTTTGGGATAAGGGTAGAGACTTCGCCACCGTCAGAAAGGTTTTGTCTATGCCCCAGGTCAACATCGTCAAGAAAACTGAAGTCCAGACCGGCGGCTTCTCCAAAGAATCTATCTTGCCAAAGAGAAACTCGGACAAGTTGATCGCCAGAAAGAAGGACTGGGACCCAAAGAAATACGGTGGTTTCGACTCTCCAACCGTCGCTTACTCTGTTTTGGTTGTCGCTAAGGTCGAGAAGGGCAAGTCTAAGAAGTTGAAGTCCGTCAAAGAGTTGTTGGGCATCACCATCATGGAAAGATCGTCCTTCGAGAAGAACCCAATCGACTTCTTGGAAGCCAAGGGCTACAAAGAGGTGAAGAAGGACTTGATCATCAAGTTGCCCAAGTACTCGTTGTTCGAGTTGGAGAACGGTAGAAAGAGAATGTTGGCTTCCGCTGGTGAATTGCAGAAGGGTAACGAATTGGCCTTGCCCTCCAAGTACGTCAACTTCTTGTACTTGGCCTCCCACTACGAAAAGTTGAAGGGTTCCCCTGAAGATAACGAGCAGAAGCAGTTGTTCGTCGAGCAGCATAAGCACTACTTGGACGAAATCATCGAGCAGATCTCCGAGTTCTCTAAGAGAGTCATCTTGGCTGACGCCAACTTGGACAAGGTCTTGTCTGCTTACAACAAGCACAGAGACAAGCCAATCAGAGAGCAGGCCGAAAACATCATCCACTTGTTCACTTTGACCAACTTGGGTGCTCCAGCTGCTTTCAAGTACTTCGACACTACCATCGACAGAAAGAGATACACCTCCACCAAAGAAGTCTTGGACGCTACCTTGATCCACCAGTCTATCACTGGCTTGTACGAAACCAGAATCGACTTGTCTCAATTGGGTGGTGAC
Protein Sequence
DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD
SV40 NLS — nuclear localization signal of SV40 (si… 9833..9854 Fwd
DNA Sequence
CCTAAGAAGAAAAGAAAAGTT
Protein Sequence
PKKKRKV
Gem1 (1-110) — degron consisting of residues 1-110 of … 9860..10190 Fwd
DNA Sequence
ATGAACCCATCTATGAAGCAGAAGCAAGAAGAGATCAAAGAAAACATCAAGAACTCGTCTGTCCCTAGAAGAACTTTGAAGATGATCCAACCATCTGCCTCTGGTTCTTTGGTTGGTAGAGAAAACGAATTGTCTGCTGGTTTGTCTAAGAGAAAGCATAGAAACGACCACTTGACTTCTACTACCTCTTCTCCAGGTGTTATCGTTCCAGAATCTTCTGAAAACAAGAACCTTGGTGGTGTCACTCAAGAATCTTTCGACTTGATGATCAAAGAGAACCCTTCTTCTCAGTACTGGAAAGAGGTTGCTGAAAAGAGAAGAAAGGCTTTG
Protein Sequence
MNPSMKQKQEEIKENIKNSSVPRRTLKMIQPSASGSLVGRENELSAGLSKRKHRNDHLTSTTSSPGVIVPESSENKNLGGVTQESFDLMIKENPSSQYWKEVAEKRRKAL
yeGFP — yeast-enhanced green fluorescent protein 11126..11843 Fwd
DNA Sequence
ATGTCTAAAGGTGAAGAATTATTCACTGGTGTTGTCCCAATTTTGGTTGAATTAGATGGTGATGTTAATGGTCACAAATTTTCTGTCTCCGGTGAAGGTGAAGGTGATGCTACTTACGGTAAATTGACCTTAAAATTTATTTGTACTACTGGTAAATTGCCAGTTCCATGGCCAACCTTAGTCACTACTTTCGGTTATGGTGTTCAATGTTTTGCTAGATACCCAGATCATATGAAACAACATGACTTTTTCAAGTCTGCCATGCCAGAAGGTTATGTTCAAGAAAGAACTATTTTTTTCAAAGATGACGGTAACTACAAGACCAGAGCTGAAGTCAAGTTTGAAGGTGATACCTTAGTTAATAGAATCGAATTAAAAGGTATTGATTTTAAAGAAGATGGTAACATTTTAGGTCACAAATTGGAATACAACTATAACTCTCACAATGTTTACATCATGGCTGACAAACAAAAGAATGGTATCAAAGTTAACTTCAAAATTAGACACAACATTGAAGATGGTTCTGTTCAATTAGCTGACCATTATCAACAAAATACTCCAATTGGTGATGGTCCAGTCTTGTTACCAGACAACCATTACTTATCCACTCAATCTGCCTTATCCAAAGATCCAAACGAAAAGAGAGACCACATGGTCTTGTTAGAATTTGTTACTGCTGCTGGTATTACCCATGGTATGGATGAATTGTACAAATAA
Protein Sequence
MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYK
promoter 5
Label Gene Product Position Strand
AmpR promoter bla — 1620..1725 Rev
DNA Sequence
CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
TRP1 promoter S. cerevisiae TRP1 — 2523..2804 Fwd
DNA Sequence
ACGACATTACTATATATATAATATAGGAAGCATTTAATAGAACAGCATCGTAATATATGTGTACTTTGCAGTTATGACGCCAGATGGCAGTAGTGGAAGATATTCTTTATTGAAAAATAGCTTGTCACCTTACGTACAATCTTGATCCGGAGCTTTTCTTTTTTTGCCGATTAAGAATTAATTCGGTCGAAAAAAGAAAAGGAGAGGGCCAAGAGGGAGGGCATTGGTGACTATTGAGCACGTGAGTATACGTGATTAAGCACACAAAGGCAGCTTGGAGT
T7 promoter — — 4203..4222 Fwd
DNA Sequence
TAATACGACTCACTATAGG
T3 promoter — — 14972..14991 Rev
DNA Sequence
AATTAACCCTCACTAAAGG
lac promoter — — 15059..15090 Rev
DNA Sequence
TTTACACTTTATGCTTCCGGCTCGTATGTTG
protein_bind 2
Label Gene Product Position Strand
lac operator — — 15035..15052 Fwd
DNA Sequence
TTGTTATCCGCTCACAA
CAP binding site — — 15104..15126 Fwd
DNA Sequence
ATGAGTGAGCTAACTCACATTA
rep_origin 2
Label Gene Product Position Strand
ori — — 0..589 Rev
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA
f1 ori — — 3581..4037 Rev
DNA Sequence
ACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGCTTACAATTT