FRP2372

127578 Synthetic Yeast Expression

Plasmid Information

Purpose

Repressor plasmid for WTC 846. Encodes TetR and TetR-Tup1, Lys2 selection marker

Depositor

Joerg Stelling

Article

Azizoglu et al Elife. 2021 Aug 3;10. pii: 69549. doi: 1

Insert

PRnr2_TetR-NLS-TUP1_tCyc1_PtetO7.1_TetR-NLS_tCyc1

Available Since

Aug. 13, 2019

Availability

Available

Sequence Information

Length

13,139 bp

Topology

circular

Molecule Type

other DNA

View on AddGene.org

Features

CDS 4

Label	Gene	Product	Position	Strand
LYS2	S. cerevisiae LYS2	alpha-aminoadipate reductase, required …	373..4552	Fwd
DNA Sequence ATGACTAACGAAAAGGTCTGGATAGAGAAGTTGGATAATCCAACTCTTTCAGTGTTACCACATGACTTTTTACGCCCACAACAAGAACCTTATACGAAACAAGCTACATATTCGTTACAGCTACCTCAGCTCGATGTGCCTCATGATAGTTTTTCTAACAAATACGCTGTCGCTTTGAGTGTATGGGCTGCATTGATATATAGAGTAACTGGTGACGATGATATTGTTCTTTATATTGCGAATAACAAAATCTTAAGATTCAATATTCAACCAACGTGGTCATTTAATGAGCTGTATTCTACAATTAACAATGAGTTGAACAAGCTCAATTCTATTGAGGCCAATTTTTCCTTTGACGAGCTAGCTGAAAAAATTCAAAGTTGCCAAGACTTGGAAAGGACCCCTCAGTTGTTCCGTTTGGCCTTTTTGGAAAACCAAGATTTCAAATTAGACGAGTTCAAGCATCATTTAGTGGACTTTGCTTTGAATTTGGATACCAGTAATAATGCGCATGTTTTGAACTTAATTTATAACAGCTTACTGTATTCGAATGAAAGAGTAACCATTGTTGCGGACCAATTTACTCAATATTTGACTGCTGCGCTAAGCGATCCATCCAATTGCATAACTAAAATCTCTCTGATCACCGCATCATCCAAGGATAGTTTACCTGATCCAACTAAGAACTTGGGCTGGTGCGATTTCGTGGGGTGTATTCACGACATTTTCCAGGACAATGCTGAAGCCTTCCCAGAGAGAACCTGTGTTGTGGAGACTCCAACACTAAATTCCGACAAGTCCCGTTCTTTCACTTATCGCGACATCAACCGCACTTCTAACATAGTTGCCCATTATTTGATTAAAACAGGTATCAAAAGAGGTGATGTAGTGATGATCTATTCTTCTAGGGGTGTGGATTTGATGGTATGTGTGATGGGTGTCTTGAAAGCTGGCGCAACCTTTTCAGTTATCGACCCTGCATATCCCCCAGCCAGACAAACCATTTACTTAGGTGTTGCTAAACCACGTGGGTTGATTGTTATTAGAGCTGCTGGACAATTGGATCAATTAGTAGAAGATTACATCAATGATGAATTGGAGATTGTTTCAAGAATCAATTCCATCGCTATTCAAGAAAATGGTACTATTGAAGGTGGCAAATTGGACAATGGCGAGGATGTTTTGGCTCCATACGATCACTACAAAGACACCAGAACAGGTGTTGTAGTTGGACCAGATTCCAACCCAACCCTATCTTTCACATCTGGTTCCGAAGGTATTCCTAAGGGTGTTCTTGGTAGACATTTTTCCTTGGCTTATTATTTCAATTGGATGTCCAAAAGGTTCAACTTAACAGAAAATGATAAATTCACAATGCTGAGCGGTATTGCACATGATCCAATTCAAAGAGATATGTTTACACCATTATTTTTAGGTGCCCAATTGTATGTCCCTACTCAAGATGATATTGGTACACCGGGCCGTTTAGCGGAATGGATGAGTAAGTATGGTTGCACAGTTACCCATTTAACACCTGCCATGGGTCAATTACTTACTGCCCAAGCTACTACACCATTCCCTAAGTTACATCATGCGTTCTTTGTGGGTGACATTTTAACAAAACGTGATTGTCTGAGGTTACAAACCTTGGCAGAAAATTGCCGTATTGTTAATATGTACGGTACTACTGAAACACAGCGTGCAGTTTCTTATTTCGAAGTTAAATCAAAAAATGACGATCCAAACTTTTTGAAAAAATTGAAAGATGTCATGCCTGCTGGTAAAGGTATGTTGAACGTTCAGCTATTAGTTGTTAACAGGAACGACCGTACTCAAATATGTGGTATTGGCGAAATAGGTGAGATTTATGTTCGTGCAGGTGGTTTGGCCGAAGGTTATAGAGGATTACCAGAATTGAATAAAGAAAAATTTGTGAACAACTGGTTTGTTGAAAAAGATCACTGGAATTATTTGGATAAGGATAATGGTGAACCTTGGAGACAATTCTGGTTAGGTCCAAGAGATAGATTGTACAGAACGGGTGATTTAGGTCGTTATCTACCAAACGGTGACTGTGAATGTTGCGGTAGGGCTGATGATCAAGTTAAAATTCGTGGGTTCAGAATCGAATTAGGAGAAATAGATACGCACATTTCCCAACATCCATTGGTAAGAGAAAACATTACTTTAGTTCGCAAAAATGCCGACAATGAGCCAACATTGATCACATTTATGGTCCCAAGATTTGACAAGCCAGATGACTTGTCTAAGTTCCAAAGTGATGTTCCAAAGGAGGTTGAAACTGACCCTATAGTTAAGGGCTTAATCGGTTACCATCTTTTATCCAAGGACATCAGGACTTTCTTAAAGAAAAGATTGGCTAGCTATGCTATGCCTTCCTTGATTGTGGTTATGGATAAACTACCATTGAATCCAAATGGTAAAGTTGATAAGCCTAAACTTCAATTCCCAACTCCCAAGCAATTAAATTTGGTAGCTGAAAATACAGTTTCTGAAACTGACGACTCTCAGTTTACCAATGTTGAGCGCGAGGTTAGAGACTTATGGTTAAGTATATTACCTACCAAGCCAGCATCTGTATCACCAGATGATTCGTTTTTCGATTTAGGTGGTCATTCTATCTTGGCTACCAAAATGATTTTTACCTTAAAGAAAAAGCTGCAAGTTGATTTACCATTGGGCACAATTTTCAAGTATCCAACGATAAAGGCCTTTGCCGCGGAAATTGACAGAATTAAATCATCGGGTGGATCATCTCAAGGTGAGGTCGTCGAAAATGTCACTGCAAATTATGCGGAAGACGCCAAGAAATTGGTTGAGACGCTACCAAGTTCGTACCCCTCTCGTGAATATTTTGTTGAACCTAATAGTGCCGAAGGAAAAACAACAATTAATGTGTTTGTTACTGGTGTCACAGGATTTCTGGGCTCCTACATCCTTGCAGATTTGTTAGGACGTTCTCCAAAGAACTACAGTTTCAAAGTGTTTGCCCACGTCAGGGCCAAGGATGAAGAAGCTGCATTTGCAAGATTACAAAAGGCAGGTATCACCTATGGTACTTGGAACGAAAAATTTGCCTCAAATATTAAAGTTGTATTAGGCGATTTATCTAAAAGCCAATTTGGTCTTTCAGATGAGAAGTGGATGGATTTGGCAAACACAGTTGATATAATTATCCATAATGGTGCGTTAGTTCACTGGGTTTATCCATACGCCAAATTGAGAGATCCAAATGTTATTTCAACTATCAATGTTATGAGCTTAGCCGCCGTCGGCAAGCCAAAGTTCTTTGACTTTGTTTCCTCCACTTCTACTCTTGACACTGAATACTACTTTAATTTGTCAGATAAACTTGTTAGCGAAGGGAAGCCAGGCATTTTAGAATCAGACGATTTAATGAACTCTGCAAGCGGGCTCACTGGTGGATATGGTCAGTCCAAATGGGCTGCTGAGTACATCATTAGACGTGCAGGTGAAAGGGGCCTACGTGGGTGTATTGTCAGACCAGGTTACGTAACAGGTGCCTCTGCCAATGGTTCTTCAAACACAGATGATTTCTTATTGAGATTTTTGAAAGGTTCAGTCCAATTAGGTAAGATTCCAGACATCGAAAATTCCGTGAATATGGTTCCAGTAGATCATGTTGCTCGTGTTGTTGTTGCTACGTCTTTGAATCCTCCCAAAGAAAATGAATTGGCCGTTGCTCAAGTAACGGGTCACCCAAGAATATTATTCAAAGACTACTTGTATACTTTACACGATTATGGTTACGATGTCGAAATCGAAAGCTATTCTAAATGGAAGAAATCATTGGAGGCGTCTGTTATTGACAGGAATGAAGAAAATGCGTTGTATCCTTTGCTACACATGGTCTTAGACAACTTACCTGAAAGTACCAAAGCTCCGGAACTAGACGATAGGAACGCCGTGGCATCTTTAAAGAAAGACACCGCATGGACAGGTGTTGATTGGTCTAATGGAATAGGTGTTACTCCAGAAGAGGTTGGTATATATATTGCATTTTTAAACAAGGTTGGATTTTTACCTCCACCAACTCATAATGACAAACTTCCACTGCCAAGTATAGAACTAACTCAAGCGCAAATAAGTCTAGTTGCTTCAGGTGCTGGTGCTCGTGGAAGCTCCGCAGCAGCTTAA Protein Sequence MTNEKVWIEKLDNPTLSVLPHDFLRPQQEPYTKQATYSLQLPQLDVPHDSFSNKYAVALSVWAALIYRVTGDDDIVLYIANNKILRFNIQPTWSFNELYSTINNELNKLNSIEANFSFDELAEKIQSCQDLERTPQLFRLAFLENQDFKLDEFKHHLVDFALNLDTSNNAHVLNLIYNSLLYSNERVTIVADQFTQYLTAALSDPSNCITKISLITASSKDSLPDPTKNLGWCDFVGCIHDIFQDNAEAFPERTCVVETPTLNSDKSRSFTYRDINRTSNIVAHYLIKTGIKRGDVVMIYSSRGVDLMVCVMGVLKAGATFSVIDPAYPPARQTIYLGVAKPRGLIVIRAAGQLDQLVEDYINDELEIVSRINSIAIQENGTIEGGKLDNGEDVLAPYDHYKDTRTGVVVGPDSNPTLSFTSGSEGIPKGVLGRHFSLAYYFNWMSKRFNLTENDKFTMLSGIAHDPIQRDMFTPLFLGAQLYVPTQDDIGTPGRLAEWMSKYGCTVTHLTPAMGQLLTAQATTPFPKLHHAFFVGDILTKRDCLRLQTLAENCRIVNMYGTTETQRAVSYFEVKSKNDDPNFLKKLKDVMPAGKGMLNVQLLVVNRNDRTQICGIGEIGEIYVRAGGLAEGYRGLPELNKEKFVNNWFVEKDHWNYLDKDNGEPWRQFWLGPRDRLYRTGDLGRYLPNGDCECCGRADDQVKIRGFRIELGEIDTHISQHPLVRENITLVRKNADNEPTLITFMVPRFDKPDDLSKFQSDVPKEVETDPIVKGLIGYHLLSKDIRTFLKKRLASYAMPSLIVVMDKLPLNPNGKVDKPKLQFPTPKQLNLVAENTVSETDDSQFTNVEREVRDLWLSILPTKPASVSPDDSFFDLGGHSILATKMIFTLKKKLQVDLPLGTIFKYPTIKAFAAEIDRIKSSGGSSQGEVVENVTANYAEDAKKLVETLPSSYPSREYFVEPNSAEGKTTINVFVTGVTGFLGSYILADLLGRSPKNYSFKVFAHVRAKDEEAAFARLQKAGITYGTWNEKFASNIKVVLGDLSKSQFGLSDEKWMDLANTVDIIIHNGALVHWVYPYAKLRDPNVISTINVMSLAAVGKPKFFDFVSSTSTLDTEYYFNLSDKLVSEGKPGILESDDLMNSASGLTGGYGQSKWAAEYIIRRAGERGLRGCIVRPGYVTGASANGSSNTDDFLLRFLKGSVQLGKIPDIENSVNMVPVDHVARVVVATSLNPPKENELAVAQVTGHPRILFKDYLYTLHDYGYDVEIESYSKWKKSLEASVIDRNEENALYPLLHMVLDNLPESTKAPELDDRNAVASLKKDTAWTGVDWSNGIGVTPEEVGIYIAFLNKVGFLPPPTHNDKLPLPSIELTQAQISLVASGAGARGSSAAA
AmpR	bla	beta-lactamase	5053..5914	Fwd
DNA Sequence ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACCATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA Protein Sequence MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
SV40 NLS	—	nuclear localization signal of SV40 (si…	7622..7643	Rev
DNA Sequence CCTAAGAAGAAGAGAAAGGTT Protein Sequence PKKKRKV
SV40 NLS	—	nuclear localization signal of SV40 (si…	11637..11658	Rev
DNA Sequence CCCAAGAAAAAGAGAAAGGTG Protein Sequence PKKKRKV

promoter 4

Label	Gene	Product	Position	Strand
LYS2 promoter	S. cerevisiae LYS2	—	133..373	Fwd
DNA Sequence AATTGCACTTGCAATTACATAAAAAATTCCGGCGGTTTTTCGCGTGTGACTCAATGTCGAAATACCTGCCTAATGAACATGAACATCGCCCAAATGTATTTGAAGACCCGCTGGGAGAAGTTCAAGATATATAAGTAACAAGCAGCCAATAGTATAAAAAAAAATCTGAGTTTATTACCTTTCCTGGAATTTCAGTGAAAAACTGCTAATTATAGAGACATATCACAGAGTTACTCACTA
AmpR promoter	bla	—	4948..5053	Fwd
DNA Sequence CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
T3 promoter	—	—	7286..7305	Fwd
DNA Sequence AATTAACCCTCACTAAAGG
T7 promoter	—	—	13114..13133	Rev
DNA Sequence TAATACGACTCACTATAGG

protein_bind 1

Label	Gene	Product	Position	Strand
lac operator	—	—	7225..7242	Fwd
DNA Sequence TTGTGAGCGGATAACAA

rep_origin 1

Label	Gene	Product	Position	Strand
ori	—	—	6092..6681	Fwd
DNA Sequence TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA

terminator 2

Label	Gene	Product	Position	Strand
CYC1 terminator	S. cerevisiae CYC1	—	7390..7580	Rev
DNA Sequence ATCCGCTCTAACCGAAAAGGAAGGAGTTAGACAACCTGAAGTCTAGGTCCCTATTTATTTTTTTATAGTTATGTTAGTATTAAGAACGTTATTTATATTTCAAATTTTTCTTTTTTTTCTGTACAGACGCGTGTACGCATGTAACATTATACTGAAAACCTTGCTTGAGAAGGTTTTGGGACGCTCGAAG
CYC1 terminator	S. cerevisiae CYC1	—	9076..9326	Rev
DNA Sequence TCATGTAATTAGTTATGTCACGCTTACATTCACGCCCTCCTCCCACATCCGCTCTAACCGAAAAGGAAGGAGTTAGACAACCTGAAGTCTAGGTCCCTATTTATTTTTTTTAATAGTTATGTTAGTATTAAGAACGTTATTTATATTTCAAATTTTTCTTTTTTTTCTGTACAAACGCGTGTACGCATGTAACATTATACTGAAAACCTTGCTTGAGAAGGTTTTGGGACGCTCGAAGGCTTTAATTTGC