LOCUS       sequence-462052-        7755 bp    DNA     circular SYN 15-APR-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 7755)
  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 7755)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pMT1981
FEATURES             Location/Qualifiers
     source          1..7755
                     /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(4692..4694,4695..4697,4698..5408)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     terminator      5486..5732
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             6129..7151
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     primer_bind     complement(7361..7378)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7361..7377)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7370..7392)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(7586..7605)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7705..7727
                     /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 cgccgctagc catggttgtg cgacagattc actgtgaaag actgttcatt
     2281 atacccacgt ttcactggga gatgtaagcc ttaggtgttt taccctgatt agataataca
     2341 ataaccaaca gaaatacgag aatctaaact aatttcgatg attcattttt ctttttaccg
     2401 cgctgcctct tttggcaatt ctttcaccta tattctacct tctctttcct tttgttctaa
     2461 acttattacc agctatctat gtcgaatcaa gaagaaggac ttaaactgtg gggtggcagg
     2521 tttactgggg ctactgaccc cttgatggat ttgtataacg cttccttacc ttacgacaag
     2581 aaaatgtaca aggtggattt agaaggaaca aaagtttaca ctgagggcct ggagaaaatt
     2641 aatttgctaa ctaaagacga actaagtgag attcatcgtg gtctcaaatt gattgaagca
     2701 gagtgggcag aagggaagtt tgttgagaag ccaggggatg aggatattca cactgctaat
     2761 gaacgtcgct tgggtgagtt gattggtcgt ggaatctctg gtaaggttca taccggaagg
     2821 tctagaaatg atcaagttgc cactgatatg cggttgtatg tcagagacaa tctaactcag
     2881 ttggctgact atctgaagca gttcattcaa gtaatcatca agagagctga acaggaaata
     2941 gacgtcttga tgcccggtta tactcacttg caaagagctc aaccaatcag atggtctcac
     3001 tggttgagca tgtatgctac ctatttcact gaagattatg agagactgaa tcaaatcgtt
     3061 aaaaggttga acaaatcccc attgggagct ggagctttgg ctggtcatcc ttatggaatt
     3121 gatcgtgaat acattgctga gagattaggg tttgattctg ttattggtaa ttctttggcc
     3181 gctgtttcag acagagattt tgtagtcgaa accatgttct ggtcttcgtt gtttatgaat
     3241 catatttctc gattctcaga agatttgatc atttactcca ctggagagtt tggatttatc
     3301 aagttggcag atgcttattc tactggatct tctctgatgc ctcaaaaaaa aaacccagac
     3361 tctttggagt tattgagggg taaatctggt agatgttttg gggccttggc tggtttcctc
     3421 atgtctatta agtccattcc gtcaacctat aacaaagata tgcaagagga taaggagcct
     3481 ttatttgata ctctaatcac tgtagagcac tcgattttga tagcatccgg tgtagtttct
     3541 accttgaaca ttgatgccga acgaatgaag aatgctctaa ctatggatat gctggctaca
     3601 gatcttgccg actatttagt tagaagggga gttccattca gagaaactca ccacatttct
     3661 ggtgaatgtg tcagacaagc cgaggagttg aacctttctg gtattgatca gttgtccctc
     3721 gaacaattga aatccattga ctcccgtttt gaggctgatg tggcttcaac gtttgacttt
     3781 gaagccagtg ttgaaaaaag aactgccacc ggaggaactt ctaagactgc tgttttaaag
     3841 caattggatg cactgaatga aaagctagag tcttgactgc aggtcgactc tagaggatcc
     3901 atgatacgaa acgtaccgta tcgttaaggt gaatgatacg aaacgtaccg tatcgttaag
     3961 gtcaatgata cgaaacgtac cgtatcgtta aggtcaatga tacgaaacgt accgtatcgt
     4021 taaggtgtat gatacgaaac gtaccgtatc gttaaggtag atgatacgaa acgtaccgta
     4081 tcgttaaggt ggcgttgtga tccggagatc catgatacga aacgtaccgt atcgttaagg
     4141 tgaatgatac gaaacgtacc gtatcgttaa ggtcaatgat acgaaacgta ccgtatcgtt
     4201 aaggtcaatg atacgaaacg taccgtatcg ttaaggtgta tgatacgaaa cgtaccgtat
     4261 cgttaaggta gatgatacga aacgtaccgt atcgttaagg tggcgttgtg atccggagat
     4321 ccatgatacg aaacgtaccg tatcgttaag gtgaatgata cgaaacgtac cgtatcgtta
     4381 aggtcaatga tacgaaacgt accgtatcgt taaggtcaat gatacgaaac gtaccgtatc
     4441 gttaaggtgt atgatacgaa acgtaccgta tcgttaaggt agatgatacg aaacgtaccg
     4501 tatcgttaag gtggcgttca gcaatatata aacagaagga agctgccctg tcttaaacct
     4561 ttttttttat catcattatt agcttacttt cataattgcg actggttcca attgacaagc
     4621 ttttgatttt aacgactttt aacgacaact tgagaagatc aaaaaacaac taattattcg
     4681 aaacgactag tatggtttct aagggtgaag agttgttcac tggtgttgtt ccaatcttgg
     4741 ttgagttgga cggtgacgtt aacggacaca agttctctgt ttctggtgaa ggtgagggtg
     4801 acgctactta cggaaagttg actttgaagt tcatctgtac tactggtaag ttgccagttc
     4861 catggccaac tttggttact actttgactt acggtgttca gtgtttctcc agatacccag
     4921 accacatgaa gcagcacgat ttcttcaagt ctgctatgcc agagggttac gttcaagaga
     4981 gaactatctt cttcaaggac gacggtaact acaagactag agctgaggtt aagttcgagg
     5041 gtgacacatt ggttaacaga atcgagttga agggtatcga cttcaaagag gacggaaaca
     5101 tcttgggtca caagttggag tacaactaca actcccacaa cgtttacatc atggctgaca
     5161 agcagaagaa cggtatcaag gttaacttca agatcagaca caacatcgag gacggttccg
     5221 ttcaattggc tgaccactac caacagaaca ctccaattgg tgacggtcca gttttgttgc
     5281 cagacaacca ctacttgtcc actcaatccg ctttgtccaa ggacccaaac gagaagagag
     5341 atcacatggt tttgttggag ttcgttactg ctgctggtat cactttgggt atggacgagt
     5401 tgtacaagta actcgagcct tagacatgac tgttcctcag ttcaagttgg gcacttacga
     5461 gaagaccggt cttgctagat tctaatcaag aggatgtcag aatgccattt gcctgagaga
     5521 tgcaggcttc atttttgata cttttttatt tgtaacctat atagtatagg attttttttg
     5581 tcattttgtt tcttctcgta cgagcttgct cctgatcagc ctatctcgca gctgatgaat
     5641 atcttgtggt aggggtttgg gaaaatcatt cgagtttgat gtttttcttg gtatttccca
     5701 ctcctcttca gagtacagaa gattaagtga gacgttcgtt tgtgcaagct tagatccccg
     5761 ggtaccgagc tcgaattcgc cgtaccactt caaaacaccc aagcacagca tactaaattt
     5821 cccctctttc ttcctctagg gtgtcgttaa ttacccgtac taaaggtttg gaaaagaaaa
     5881 aagagaccgc ctcgtttctt tttcttcgtc gaaaaaggca ataaaaattt ttatcacgtt
     5941 tctttttctt gaaaattttt ttttttgatt tttttctctt tcgatgacct cccattgata
     6001 tttaagttaa taaacggtct tcaatttctc aagtttcagt ttcatttttc ttgttctatt
     6061 acaacttttt ttacttcttg ctcattagaa agaaagcata gcaatctaat ctaagtttta
     6121 attacaaaat gaagaagcca gagttgactg ctacttccgt tgagaagttc ttgatcgaga
     6181 agttcgactc cgtttccgac ttgatgcaat tgtctgaagg tgaagagtcc agagctttct
     6241 cctttgacgt tggtggtaga ggttacgttt tgagagttaa ttcctgtgct gacggtttct
     6301 acaaggacag atacgtttac agacacttcg cttccgctgc tttgccaatt ccagaggttt
     6361 tggacattgg tgagttctcc gagtccttga cttactgtat ctccagaaga gctcagggag
     6421 ttacattgca agacttgcca gaaactgagt tgccagctgt tttgcaacca gttgctgaag
     6481 ctatggacgc tattgctgct gctgacttgt ctcaaacttc tggttttggt ccattcggtc
     6541 cacagggtat tggtcagtac actacttgga gagacttcat ctgtgctatc gctgacccac
     6601 atgtttacca ctggcaaact gttatggacg atactgtttc cgcttccgtt gctcaggctt
     6661 tggacgaatt gatgttgtgg gctgaagatt gtcctgaggt tagacatttg gttcacgctg
     6721 acttcggttc caacaacgtt ttgactgaca acggtagaat cactgctgtt atcgactggt
     6781 ccgaagctat gttcggtgac tctcaatacg aggttgctaa catcttcttt tggagaccat
     6841 ggttggcttg tatggaacag cagactagat acttcgagag aagacaccca gaattggctg
     6901 gttccccaag attgagagct tacatgttga gaatcggttt ggaccagttg taccagtcct
     6961 tggttgatgg taacttcgat gatgctgctt gggctcaagg tagatgtgac gctatcgtta
     7021 gatccggtgc tggtactgtt ggtagaactc agattgctag aagatccgct gctgtttgga
     7081 ctgacggttg tgttgaagtt ttggctgact ccggtaacag aagaccatcc actagaccaa
     7141 gagctaaaga ggttggtaga gtttaagtat taagaacgtt atttatattt caaatttttc
     7201 ttttttttct ctacagacgc tgtacgcatg taacattata ctgaaaacct tgcttgagaa
     7261 ggttttggga cgctcgaaga tccagctgca ttaatgaatc ggccaacgcg cggggagagg
     7321 cggtttgcgt attgggcgct cttccgcttc ctcggaattc actggccgtc gttttacaac
     7381 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     7441 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     7501 gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     7561 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     7621 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     7681 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     7741 caccgaaacg cgcga
//
