LOCUS       sequence-386220-        5881 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid used for the C-terminal tagging of Ugp1 with mCherry and the
            auxin-inducible degron in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5881)
  AUTHORS   Takhaveev V, Ozsezen S, Smith EN, Zylstra A, Chaillet ML, Chen H, 
            Papagiannakis A, Milias-Argeitis A, Heinemann M
  TITLE     Temporal segregation of biosynthetic processes is responsible for 
            metabolic oscillations during the budding yeast cell cycle.
  JOURNAL   Nat Metab. 2023 Feb;5(2):294-313. doi: 10.1038/s42255-023-00741-x. 
            Epub 2023 Feb 27.
  PUBMED    36849832
REFERENCE   2  (bases 1 to 5881)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s42255-023-00741-x"; journalName: "Nat Metab"; date: 
            "2023-02"; volume: "5"; issue: "2"; pages: "294-313"
COMMENT     Sequence Label: pUGP1.1
FEATURES             Location/Qualifiers
     source          1..5881
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             507..1214
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(652..670)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     920..939
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1257..1388
                     /codon_start=1
                     /product="minimal degron from the auxin-responsive IAA17 
                     protein of Arabidopsis thaliana (Morawska and Ulrich, 
                     2013)"
                     /label=AID*
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="PKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEAAAFVK"
     terminator      1436..1625
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     gene            complement(1680..2799)
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     terminator      complement(1680..1877)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(1883..2455)
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     promoter        complement(2456..2799)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     primer_bind     3356..3377
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3531..3550)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3650..3672
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3710..3728)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3796..3900
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3901..3969,3970..4761)
                     /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(4119..4138)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4932..5520
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5421..5440
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5674..5691
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5808..5829
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tgatcgaatc gagcaatttg gagatggaaa tcattccaaa ccaaaaaact ataacaagag
       61 acggtcatga aattaatgtc ttacaattag aaaccgcttg tggtgctgct atcaggcatt
      121 ttgatggtgc tcacggtgtt gtcgttccaa gatcaagatt cttgcctgtc aagacctgtt
      181 ccgatttgtt gctggttaaa tcagatctat tccgtctgga acacggttct ttgaagttag
      241 acccatcccg ttttggtcca aacccattaa tcaagttggg ctcgcatttc aaaaaggttt
      301 ctggttttaa cgcaagaatc cctcacatcc caaaaatcgt cgagctagat catttgacca
      361 tcactggtaa cgtcttttta ggtaaagatg tcactttgag gggtactgtc atcatcgttt
      421 gctccgacgg tcataaaatc gatattccaa acggctccat attggaaaat gttgtcgtta
      481 ctggtaattt gcaaatcttg gaacatatgg tgagcaaggg cgaggaggat aacatggcca
      541 tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt
      601 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
      661 aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt
      721 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct
      781 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
      841 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
      901 ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct
      961 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc
     1021 tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg
     1081 tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg
     1141 actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg
     1201 acgagctgta caaggtcgac ggtgcaggcg ccggggctgg tgctggggca ggtaccccta
     1261 aagatccagc caaacctccg gccaaggcac aagttgtggg atggccaccg gtgagatcat
     1321 accggaagaa cgtgatggtt tcctgccaaa aatcaagcgg tggcccggag gcggcggcgt
     1381 tcgtgaaggt atcacccggg taactcgagt aagcttggta ccgcggctag ctaagatccg
     1441 ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt tattttttta
     1501 tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca
     1561 gacgcgtgta cgcatgtaac attatactga aaaccttgct tgagaaggtt ttgggacgct
     1621 cgaagatcca gctcagcgta cgtcgacact ggatggcggc gttagtatcg aatcgacagc
     1681 agtatagcga ccagcattca catacgattg acgcatgata ttactttctg cgcacttaac
     1741 ttcgcatctg ggcagatgat gtcgaggcga aaaaaaatat aaatcacgct aacatttgat
     1801 taaaatagaa caactacaat ataaaaaaac tatacaaatg acaagttctt gaaaacaaga
     1861 atctttttat tgtcagtact gattaggggc agggcatgct catgtagagc gcctgctcgc
     1921 cgtccgaggc ggtgccgtcg tacagggcgg tgtccaggcc gcagagggtg aaccccatcc
     1981 gccggtacgc gtggatcgcc ggtgcgttga cgttggtgac ctccagccag aggtgcccgg
     2041 cgccccgctc gcgggcgaac tccgtcgcga gccccatcaa cgcgcgcccg accccgtgcc
     2101 cccggtgctc cggggcgacc tcgatgtcct cgacggtcag ccggcggttc cagccggagt
     2161 acgagacgac cacgaagccc gccaggtcgc cgtcgtcccc gtacgcgacg aacgtccggg
     2221 agtccgggtc gccgtcctcc ccgtcgtccg attcgtcgtc cgattcgtcg tcggggaaca
     2281 ccttggtcag gggcgggtcc accggcacct cccgcagggt gaagccgtcc ccggtggcgg
     2341 tgacgcggaa gacggtgtcg gtggtgaagg acccatccag tgcctcgatg gcctcggcgt
     2401 cccccgggac actggtgcgg taccggtaag ccgtgtcgtc aagagtggta cccatggttg
     2461 tttatgttcg gatgtgatgt gagaactgta tcctagcaag attttaaaag gaagtatatg
     2521 aaagaagaac ctcagtggca aatcctaacc ttttatattt ctctacaggg gcgcggcgtg
     2581 gggacaattc aacgcgtctg tgaggggagc gtttccctgc tcgcaggtct gcagcgagga
     2641 gccgtaattt ttgcttcgcg ccgtgcggcc atcaaaatgt atggatgcaa atgattatac
     2701 atggggatgt atgggctaaa tgtacgggcg acagtcacat catgcccctg agctgcgcac
     2761 gtcaagactg tcaaggaggg tattctgggc ctccatgtcg ctggccgggt gacccggcgg
     2821 ggacaaggca agcttttact ttcaattctc cgttaggtta tttattctac tacacgcagt
     2881 tttttttttt caatgggtta tatatctcta atatatacac atttatatgt ttagacttat
     2941 acagaagcaa aaaaaaatgc aaaagaagct tagtgttcat aattcttcac tcacgtcatg
     3001 ttatatattt gttaagcgta gcttcagtat atatagagat tttcctttgg cggggtaagg
     3061 gtctgtgaac tttagggaaa tagaataaac gtaagttaaa ggtgacaagc tcggatcgtc
     3121 atctttcaat tatcacccca aaagatattg taaaaaagca agttcaatct cgtttgttac
     3181 atttgtcccg tatttctact aattcaggca aaaaaggaaa gtcctacagc aaaagaggca
     3241 aaagtacagt atggaaatcg atggcaacac tctggtgttc ataatagtga tccttttcct
     3301 atttttctcc tcaccgggtg gtgatggtgt gtcttctcaa tatgagttca taatagtgga
     3361 ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa
     3421 gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3481 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
     3541 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     3601 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     3661 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     3721 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     3781 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3841 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     3901 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     3961 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     4021 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     4081 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     4141 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     4201 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     4261 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     4321 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     4381 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     4441 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     4501 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     4561 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     4621 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     4681 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     4741 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     4801 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     4861 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     4921 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4981 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5041 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     5101 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5161 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5221 ttaccggata aggcacagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5281 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     5341 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     5401 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     5461 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     5521 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     5581 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     5641 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     5701 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     5761 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     5821 ctcactcatt aggcacccca ggctttacac tttatgcttc cgcggctcgt atgttgtgtg
     5881 g
//
