LOCUS       sequence-327174-        5870 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrative expression of yEGFP-AID*(mini auxin inducible 
            degron)-CUP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5870)
  AUTHORS   Lu Z, Peng B, Ebert BE, Dumsday G, Vickers CE
  TITLE     Auxin-mediated protein depletion for metabolic engineering in 
            terpene-producing yeast.
  JOURNAL   Nat Commun. 2021 Feb 16;12(1):1051. doi: 10.1038/s41467-021-21313-1.
  PUBMED    33594068
REFERENCE   2  (bases 1 to 5870)
  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"; journalName: "Nat 
            Commun."; date: "2021-02-16"; volume: "12(1):1051. doi"; pages: " 
            10.1038/s41467-021-21313-1"
COMMENT     Sequence Label: pILGFP104
FEATURES             Location/Qualifiers
     source          1..5870
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             431..1141
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             1148..1279
                     /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      1472..1742
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
     primer_bind     complement(1891..1907)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1891..1907)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1904..1926)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1915..1931
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1939..1945,1946..1963,1964..1969))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1984..2005
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2122..2139)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2293..2881)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2373..2392)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3052..3843,3844..3912))
                     /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     3675..3694
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3913..4017)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4085..4103
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4141..4163)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4263..4282
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4476..4498
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4490..4507
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4491..4507
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5621..5654)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(5865..5870,1..408)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
ORIGIN
        1 ccacacacca tagcttcaaa atgtttctac tcctttttta ctcttccaga ttttctcgga
       61 ctccgcgcat cgccgtacca cttcaaaaca cccaagcaca gcatactaaa tttcccctct
      121 ttcttcctct agggtgtcgt taattacccg tactaaaggt ttggaaaaga aaaaagagac
      181 cgcctcgttt ctttttcttc gtcgaaaaag gcaataaaaa tttttatcac gtttcttttt
      241 cttgaaaatt tttttttttg atttttttct ctttcgatga cctcccattg atatttaagt
      301 taataaacgg tcttcaattt ctcaagtttc agtttcattt ttcttgttct attacaactt
      361 tttttacttc ttgctcatta gaaagaaagc atagcaatct aatctaagtt ttaattacaa
      421 aatgggatcc tctaaaggtg aagaattatt cactggtgtt gtcccaattt tggttgaatt
      481 agatggtgat gttaatggtc acaaattttc tgtctccggt gaaggtgaag gtgatgctac
      541 ttacggtaaa ttgaccttaa aatttatttg tactactggt aaattgccag ttccatggcc
      601 aaccttagtc actactttcg gttatggtgt tcaatgtttt gcgagatacc cagatcatat
      661 gaaacaacat gactttttca agtctgccat gccagaaggt tatgttcaag aaagaactat
      721 ttttttcaaa gatgacggta actacaagac cagagctgaa gtcaagtttg aaggtgatac
      781 cttagttaat agaatcgaat taaaaggtat tgattttaaa gaagatggta acattttagg
      841 tcacaaattg gaatacaact ataactctca caatgtttac atcatggctg acaaacaaaa
      901 gaatggtatc aaagttaact tcaaaattag acacaacatt gaagatggtt ctgttcaatt
      961 agctgaccat tatcaacaaa atactccaat tggtgatggt ccagtcttgt taccagacaa
     1021 ccattactta tccactcaat ctgccttatc caaagatcca aacgaaaaga gagaccacat
     1081 ggtcttgtta gaatttgtta ctgctgctgg tattacccat ggtatggatg aattgtacaa
     1141 aagatctcca aaagacccag ctaaacctcc agccaaggcc caagtcgtcg gctggccacc
     1201 agttaggtct tacagaaaga acgtaatggt tagttgtcaa aagtcctccg gtggtccaga
     1261 agctgctgcc tttgttaaga tgttcagcga attaattaac ttccaaaatg aaggtcatga
     1321 gtgccaatgc caatgtggta gctgcaaaaa taatgaacaa tgccaaaaat catgtagctg
     1381 cccaacgggg tgtaacagcg acgacaaatg cccctgcggt aacaagtctg aagaaaccaa
     1441 gaagtcatgc tgctctggga aataaactag tattgaattg aattgaaatc gatagatcaa
     1501 tttttttctt ttctctttcc ccatccttta cgctaaaata atagtttatt ttattttttg
     1561 aatatttttt atttatatac gtatatatag actattattt atcttttaat gattattaag
     1621 atttttatta aaaaaaaatt cgctcctctt ttaatgcctt tatgcagttt ttttttccca
     1681 ttcgatattt ctatgttcgg gttcagcgta ttttaagttt aataactcga aaattctgcg
     1741 ttcgttaaag ctttcgagaa ggatattatt tgctaagtaa atgcatgtat actaaactca
     1801 caaattagag cttcaattta attatatcag ttattacccg ggaatctcgg tcgtaatgat
     1861 ttaaatgcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     1921 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     1981 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     2041 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     2101 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     2161 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2221 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2281 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2341 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2401 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2461 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2521 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2581 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2641 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2701 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     2761 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2821 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2881 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     2941 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3001 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3061 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3121 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3181 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3241 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3301 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3361 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3421 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     3481 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     3541 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     3601 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     3661 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     3721 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     3781 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     3841 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     3901 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     3961 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4021 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     4081 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     4141 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     4201 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     4261 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     4321 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     4381 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     4441 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     4501 ggccagtgaa ttcatttaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga
     4561 aggaaggagc acagacttag attggtatat atacgcatat gtagtgttga agaaacatga
     4621 aattgcccag tattcttaac ccaactgcac agaacaaaaa cctgcaggaa acgaagataa
     4681 aacatgtcca caaaatcata taccagtaga gctgagactc atgcaagtcc ggttgcatcg
     4741 aaacttttac gtttaatgga tgaaaagaag accaatttgt gtgcttctct tgacgttcgt
     4801 tcgactgatg agctattgaa acttgttgaa acgttgggtc catacatttg ccttttgaaa
     4861 acacacgttg atatcttgga tgatttcagt tatgagggta ctgtcgttcc attgaaagca
     4921 ttggcagaga aatacaagtt cttgatattt gaggacagaa aattcgccga tatcggtaac
     4981 acagtcaaat tacaatatac atcgggcgtt taccgtatcg cagaatggtc tgatatcacc
     5041 aacgcccacg gggttactgg tgctggtatt gttgctggct tgaaacaagg tgcgcaagag
     5101 gtcaccaaag aaccaagggg attattgatg cttgctgaat tgtcttccaa gggttctcta
     5161 gcacacggtg aatatactaa gggtaccgtt gatattgcaa agagtgataa agatttcgtt
     5221 attgggttca ttgctcagaa cgatatggga ggaagagaag aagggtttga ttggctaatc
     5281 atgaccccag gtgtaggttt agacgacaaa ggcgatgcat tgggtcagca gtacagaacc
     5341 gtcgacgaag ttgtaagtgg tggatcagat atcatcattg ttggcagagg acttttcgcc
     5401 aagggtagag atcctaaggt tgaaggtgaa agatacagaa atgctggatg ggaagcgtac
     5461 caaaagagaa tcagcgctcc ccattaatta tacaggaaac ttaatagaac aaatcacata
     5521 tttaatctaa tagccacctg cattggcacg gtgcaacact acttcaactt catcttacaa
     5581 aaagatcacg tgatctgttg tattgggatc tctagaccta ataacttcgt atagcataca
     5641 ttatacgaag ttatattaag ggttgctcga ccagaaagcg accacccaac tttggctgat
     5701 aatagcgtat aaacaatgca tactttgtac gttcaaaata caatgcagta gatatattta
     5761 tgcatattac atataataca tatcacatag gaagcaacag gcgcgttgga cttttaattt
     5821 tcgaggaccg cgaatcctta catcacaccc aatcccccac aagtgatccc
//
