LOCUS       sequence-458296-        5891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  2,4-Diacetylphloroglucinol (DAPG)-regulatable EGFP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5891)
  AUTHORS   Tominaga M, Shima Y, Nozaki K, Ito Y, Someda M, Shoya Y, Hashii N, 
            Obata C, Matsumoto-Kitano M, Suematsu K, Matsukawa T, Hosoya K, 
            Hashiba N, Kondo A, Ishii J
  TITLE     Designing strong inducible synthetic promoters in yeasts.
  JOURNAL   Nat Commun. 2024 Dec 19;15(1):10653. doi: 
            10.1038/s41467-024-54865-z.
  PUBMED    39702268
REFERENCE   2  (bases 1 to 5891)
  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: "2024-12-19"; volume: "15(1):10653. doi"; pages: " 
            10.1038/s41467-024-54865-z"
COMMENT     Sequence Label: pMT2093
FEATURES             Location/Qualifiers
     source          1..5891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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)."
     primer_bind     2187..2209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2206..2222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(2721..2723,2724..2726,2727..3437)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     terminator      3515..3761
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             complement(4410..5204)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4521..4540)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5131..5150
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(5497..5514)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5497..5513)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5506..5528)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5722..5741)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5841..5863
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctt gcggccgcgg atcctttacc gtaggattta cttataccca
     2281 actggcgcgc ctcctttctc ttggatatag ctcatccaga tcaaagggaa tgcttaaatt
     2341 gagaaatagc tgtctcctta taatactttg tttccaattc aactgttgaa gattatttgg
     2401 ctcacttgat agttggttta acaatgccct agccctttca tcaagaaacg gtcgattttc
     2461 ctctatagcg tggatcctat gtatgatacg aaacgtaccg tatcgttaag gtagcgtcaa
     2521 aatttaactg ttctaacccc tacttgacag caatatataa acagaaggaa gctgccctgt
     2581 cttaaacctt tttttttatc atcattatta gcttactttc ataattgcga ctggttccaa
     2641 ttgacaagct tttgatttta acgactttta acgacaactt gagaagatca aaaaacaact
     2701 aattattcga aacgactagt atggtttcta agggtgaaga gttgttcact ggtgttgttc
     2761 caatcttggt tgagttggac ggtgacgtta acggacacaa gttctctgtt tctggtgaag
     2821 gtgagggtga cgctacttac ggaaagttga ctttgaagtt catctgtact actggtaagt
     2881 tgccagttcc atggccaact ttggttacta ctttgactta cggtgttcag tgtttctcca
     2941 gatacccaga ccacatgaag cagcacgatt tcttcaagtc tgctatgcca gagggttacg
     3001 ttcaagagag aactatcttc ttcaaggacg acggtaacta caagactaga gctgaggtta
     3061 agttcgaggg tgacacattg gttaacagaa tcgagttgaa gggtatcgac ttcaaagagg
     3121 acggaaacat cttgggtcac aagttggagt acaactacaa ctcccacaac gtttacatca
     3181 tggctgacaa gcagaagaac ggtatcaagg ttaacttcaa gatcagacac aacatcgagg
     3241 acggttccgt tcaattggct gaccactacc aacagaacac tccaattggt gacggtccag
     3301 ttttgttgcc agacaaccac tacttgtcca ctcaatccgc tttgtccaag gacccaaacg
     3361 agaagagaga tcacatggtt ttgttggagt tcgttactgc tgctggtatc actttgggta
     3421 tggacgagtt gtacaagtaa ctcgagcctt agacatgact gttcctcagt tcaagttggg
     3481 cacttacgag aagaccggtc ttgctagatt ctaatcaaga ggatgtcaga atgccatttg
     3541 cctgagagat gcaggcttca tttttgatac ttttttattt gtaacctata tagtatagga
     3601 ttttttttgt cattttgttt cttctcgtac gagcttgctc ctgatcagcc tatctcgcag
     3661 ctgatgaata tcttgtggta ggggtttggg aaaatcattc gagtttgatg tttttcttgg
     3721 tatttcccac tcctcttcag agtacagaag attaagtgag acgttcgttt gtgcaagctt
     3781 agatcttcta gagacaataa gaagaaaaaa aaagaaaagc ggtgggggag ggattattaa
     3841 ataaggatta tgtaacccca gggtaccgtt ctatacatat ttaaggatta tttaggacaa
     3901 tcgatgaaat cggcatcaaa ctggatggga gtatagtgtc cggataatcg gataaatcat
     3961 cttgcgagga gccgcttggt tggttggtga gaggagtgaa atatgtgtct cctcacccaa
     4021 gaatcgcgat atcagcaccc tgtgggggac actattggcc tccctcccaa accttcgatg
     4081 tggtagtgct ttattatatt gattacattg attacatagc taaaccctgc ctggttgcaa
     4141 gttgagctcc gaattccaat attagtaaaa tgcctgcaag ataacctcgg tatggcgtcc
     4201 gaccccgctt aattatttta actcctttcc aacgaggact tcgtaatttt tgattaggga
     4261 gttgagaaac ggggggtctt gatacctcct cgatttcaga tcccaccccc tctcagtccc
     4321 aagtgggacc cccctcggcc gtgaaatgcg cgcactttag tttttttcgc atgtaaacgc
     4381 cggtgtccgt catctagaga attccccgct cagaagaact cgtcaagaag gcgatagaag
     4441 gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat
     4501 tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc
     4561 gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata
     4621 ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catgcgcgcc
     4681 ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc
     4741 tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg
     4801 tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg
     4861 atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg
     4921 cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga
     4981 acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cctgcagttc attcagggca
     5041 ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg
     5101 gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc
     5161 caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatatggtt tctatattat
     5221 ctttgtacta aagagcaatt gataatgtgc gagaaaaact ggtccttata taccgtttgc
     5281 agcactccct cccgaacttt acgaaaagtc gtgcgccacc tgattttcat cacgccaaaa
     5341 acctacacgt atgactactc cgggccagtg ttcaccacga gctatatagt gttaattaat
     5401 taccttattg gttagctctg catgtaaggg tggtgtgagc cgggaattgg gtctactcta
     5461 gcgttcagta aggtgatata aagctctgta gaattcactg gccgtcgttt tacaacgtcg
     5521 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     5581 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     5641 gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     5701 ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     5761 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     5821 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     5881 gaaacgcgcg a
//
