LOCUS       sequence-327173-        5878 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrative expression of CUP1-AID* (mini auxin inducible 
            degron)-yEGFP .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5878)
  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 5878)
  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: pILGFP5V1
FEATURES             Location/Qualifiers
     source          1..5878
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             614..745
                     /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"
     CDS             755..1465
                     /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"
     terminator      1481..1751
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
     primer_bind     complement(1899..1915)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1899..1915)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1912..1934)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1923..1939
                     /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(1947..1953,1954..1971,1972..1977))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1992..2013
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2130..2147)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2301..2889)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2381..2400)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3060..3851,3852..3920))
                     /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     3683..3702
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3921..4025)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4093..4111
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4149..4171)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4271..4290
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4484..4506
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4498..4515
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4499..4515
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5629..5662)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(5873..5878,1..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 actcgagaaa atgttcagcg aattaattaa cttccaaaat gaaggtcatg agtgccaatg
      481 ccaatgtggt agctgcaaaa ataatgaaca atgccaaaaa tcatgtagct gcccaacggg
      541 gtgtaacagc gacgacaaat gcccctgcgg taacaagtct gaagaaacca agaagtcatg
      601 ctgctctggg aaaccaaaag acccagctaa acctccagcc aaggcccaag tcgtcggctg
      661 gccaccagtt aggtcttaca gaaagaacgt aatggttagt tgtcaaaagt cctccggtgg
      721 tccagaagct gctgcctttg ttaagatggg atcctctaaa ggtgaagaat tattcactgg
      781 tgttgtccca attttggttg aattagatgg tgatgttaat ggtcacaaat tttctgtctc
      841 cggtgaaggt gaaggtgatg ctacttacgg taaattgacc ttaaaattta tttgtactac
      901 tggtaaattg ccagttccat ggccaacctt agtcactact ttcggttatg gtgttcaatg
      961 ttttgcgaga tacccagatc atatgaaaca acatgacttt ttcaagtctg ccatgccaga
     1021 aggttatgtt caagaaagaa ctattttttt caaagatgac ggtaactaca agaccagagc
     1081 tgaagtcaag tttgaaggtg ataccttagt taatagaatc gaattaaaag gtattgattt
     1141 taaagaagat ggtaacattt taggtcacaa attggaatac aactataact ctcacaatgt
     1201 ttacatcatg gctgacaaac aaaagaatgg tatcaaagtt aacttcaaaa ttagacacaa
     1261 cattgaagat ggttctgttc aattagctga ccattatcaa caaaatactc caattggtga
     1321 tggtccagtc ttgttaccag acaaccatta cttatccact caatctgcct tatccaaaga
     1381 tccaaacgaa aagagagacc acatggtctt gttagaattt gttactgctg ctggtattac
     1441 ccatggtatg gatgaattgt acaaaagatc ttaaactagt attgaattga attgaaatcg
     1501 atagatcaat ttttttcttt tctctttccc catcctttac gctaaaataa tagtttattt
     1561 tattttttga atatttttta tttatatacg tatatataga ctattattta tcttttaatg
     1621 attattaaga tttttattaa aaaaaaattc gctcctcttt taatgccttt atgcagtttt
     1681 tttttcccat tcgatatttc tatgttcggg ttcagcgtat tttaagttta ataactcgaa
     1741 aattctgcgt tcgttaaagc tttcgagaag gatattattt gtaagtaaat gcatgtatac
     1801 taaactcaca aattagagct tcaatttaat tatatcagtt attacccggg aatctcggtc
     1861 gtaatgattt aaatgcatgc aagcttggcg taatcatggt catagctgtt tcctgtgtga
     1921 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     1981 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     2041 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     2101 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2161 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2221 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2281 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2341 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2401 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2461 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2521 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2581 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2641 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2701 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     2761 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     2821 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     2881 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     2941 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3001 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3061 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3121 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3181 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3241 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3301 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3361 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3421 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3481 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3541 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3601 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     3661 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     3721 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     3781 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     3841 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     3901 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     3961 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4021 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     4081 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     4141 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     4201 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     4261 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     4321 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc
     4381 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     4441 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     4501 aaaacgacgg ccagtgaatt catttaaatt tttttgattc ggtaatctcc gaacagaagg
     4561 aagaacgaag gaaggagcac agacttagat tggtatatat acgcatatgt agtgttgaag
     4621 aaacatgaaa ttgcccagta ttcttaaccc aactgcacag aacaaaaacc tgcaggaaac
     4681 gaagataaaa catgtccaca aaatcatata ccagtagagc tgagactcat gcaagtccgg
     4741 ttgcatcgaa acttttacgt ttaatggatg aaaagaagac caatttgtgt gcttctcttg
     4801 acgttcgttc gactgatgag ctattgaaac ttgttgaaac gttgggtcca tacatttgcc
     4861 ttttgaaaac acacgttgat atcttggatg atttcagtta tgagggtact gtcgttccat
     4921 tgaaagcatt ggcagagaaa tacaagttct tgatatttga ggacagaaaa ttcgccgata
     4981 tcggtaacac agtcaaatta caatatacat cgggcgttta ccgtatcgca gaatggtctg
     5041 atatcaccaa cgcccacggg gttactggtg ctggtattgt tgctggcttg aaacaaggtg
     5101 cgcaagaggt caccaaagaa ccaaggggat tattgatgct tgctgaattg tcttccaagg
     5161 gttctctagc acacggtgaa tatactaagg gtaccgttga tattgcaaag agtgataaag
     5221 atttcgttat tgggttcatt gctcagaacg atatgggagg aagagaagaa gggtttgatt
     5281 ggctaatcat gaccccaggt gtaggtttag acgacaaagg cgatgcattg ggtcagcagt
     5341 acagaaccgt cgacgaagtt gtaagtggtg gatcagatat catcattgtt ggcagaggac
     5401 ttttcgccaa gggtagagat cctaaggttg aaggtgaaag atacagaaat gctggatggg
     5461 aagcgtacca aaagagaatc agcgctcccc attaattata caggaaactt aatagaacaa
     5521 atcacatatt taatctaata gccacctgca ttggcacggt gcaacactac ttcaacttca
     5581 tcttacaaaa agatcacgtg atctgttgta ttgggatctc tagacctaat aacttcgtat
     5641 agcatacatt atacgaagtt atattaaggg ttgctcgacc agaaagcgac cacccaactt
     5701 tggctgataa tagcgtataa acaatgcata ctttgtacgt tcaaaataca atgcagtaga
     5761 tatatttatg catattacat ataatacata tcacatagga agcaacaggc gcgttggact
     5821 tttaattttc gaggaccgcg aatccttaca tcacacccaa tcccccacaa gtgatccc
//
