LOCUS       sequence-461541-        7721 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homoserine lactone (HSL)-regulatable EGFP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7721)
  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 7721)
  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: pMT1877
FEATURES             Location/Qualifiers
     source          1..7721
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(3427..3429,3430..3432,3433..4143)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     terminator      4221..4467
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             4864..5886
                     /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(6096..6113)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6096..6112)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6105..6127)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6321..6340)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6440..6462
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6500..6518)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6586..6690
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6691..6759,6760..7551)
                     /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(6909..6928)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccgctagcc atggttgtgc
     1021 gacagattca ctgtgaaaga ctgttcatta tacccacgtt tcactgggag atgtaagcct
     1081 taggtgtttt accctgatta gataatacaa taaccaacag aaatacgaga atctaaacta
     1141 atttcgatga ttcatttttc tttttaccgc gctgcctctt ttggcaattc tttcacctat
     1201 attctacctt ctctttcctt ttgttctaaa cttattacca gctatctatg tcgaatcaag
     1261 aagaaggact taaactgtgg ggtggcaggt ttactggggc tactgacccc ttgatggatt
     1321 tgtataacgc ttccttacct tacgacaaga aaatgtacaa ggtggattta gaaggaacaa
     1381 aagtttacac tgagggcctg gagaaaatta atttgctaac taaagacgaa ctaagtgaga
     1441 ttcatcgtgg tctcaaattg attgaagcag agtgggcaga agggaagttt gttgagaagc
     1501 caggggatga ggatattcac actgctaatg aacgtcgctt gggtgagttg attggtcgtg
     1561 gaatctctgg taaggttcat accggaaggt ctagaaatga tcaagttgcc actgatatgc
     1621 ggttgtatgt cagagacaat ctaactcagt tggctgacta tctgaagcag ttcattcaag
     1681 taatcatcaa gagagctgaa caggaaatag acgtcttgat gcccggttat actcacttgc
     1741 aaagagctca accaatcaga tggtctcact ggttgagcat gtatgctacc tatttcactg
     1801 aagattatga gagactgaat caaatcgtta aaaggttgaa caaatcccca ttgggagctg
     1861 gagctttggc tggtcatcct tatggaattg atcgtgaata cattgctgag agattagggt
     1921 ttgattctgt tattggtaat tctttggccg ctgtttcaga cagagatttt gtagtcgaaa
     1981 ccatgttctg gtcttcgttg tttatgaatc atatttctcg attctcagaa gatttgatca
     2041 tttactccac tggagagttt ggatttatca agttggcaga tgcttattct actggatctt
     2101 ctctgatgcc tcaaaaaaaa aacccagact ctttggagtt attgaggggt aaatctggta
     2161 gatgttttgg ggccttggct ggtttcctca tgtctattaa gtccattccg tcaacctata
     2221 acaaagatat gcaagaggat aaggagcctt tatttgatac tctaatcact gtagagcact
     2281 cgattttgat agcatccggt gtagtttcta ccttgaacat tgatgccgaa cgaatgaaga
     2341 atgctctaac tatggatatg ctggctacag atcttgccga ctatttagtt agaaggggag
     2401 ttccattcag agaaactcac cacatttctg gtgaatgtgt cagacaagcc gaggagttga
     2461 acctttctgg tattgatcag ttgtccctcg aacaattgaa atccattgac tcccgttttg
     2521 aggctgatgt ggcttcaacg tttgactttg aagccagtgt tgaaaaaaga actgccaccg
     2581 gaggaacttc taagactgct gttttaaagc aattggatgc actgaatgaa aagctagagt
     2641 cttgactgca ggtcgacagc gtgtttttta ttccttaagt tacgcaaggt cgaacataag
     2701 tacctgtagg atcgtacagg tttacgcaag gtcgaacata agtacctgta ggatcgtaca
     2761 ggtcgctatt acgccagcgg atccacctgt aggatcgtac aggtgaacct gtaggatcgt
     2821 acaggtcaac ctgtaggatc gtacaggtac gtatgtgatc cggatgataa tgttaagtta
     2881 cgcaaggtcg aacataagta cctgtaggat cgtacaggtt tacgcaaggt cgaacataag
     2941 tacctgtagg atcgtacagg tcgctattac gccagcggat ccacctgtag gatcgtacag
     3001 gtgaacctgt aggatcgtac aggtcaacct gtaggatcgt acaggtacgt atgtgatccg
     3061 gatgataatg ttaagttacg caaggtcgaa cataagtacc tgtaggatcg tacaggttta
     3121 cgcaaggtcg aacataagta cctgtaggat cgtacaggtc gctattacgc cagcggatcc
     3181 aactgtaaaa tcggacaggt gaaactgtaa aatcggacag gtcaaactgt aaaatcggac
     3241 aggtacgtat gtgaagcaat atataaacag aaggaagctg ccctgtctta aacctttttt
     3301 tttatcatca ttattagctt actttcataa ttgcgactgg ttccaattga caagcttttg
     3361 attttaacga cttttaacga caacttgaga agatcaaaaa acaactaatt attcgaaacg
     3421 actagtatgg tttctaaggg tgaagagttg ttcactggtg ttgttccaat cttggttgag
     3481 ttggacggtg acgttaacgg acacaagttc tctgtttctg gtgaaggtga gggtgacgct
     3541 acttacggaa agttgacttt gaagttcatc tgtactactg gtaagttgcc agttccatgg
     3601 ccaactttgg ttactacttt gacttacggt gttcagtgtt tctccagata cccagaccac
     3661 atgaagcagc acgatttctt caagtctgct atgccagagg gttacgttca agagagaact
     3721 atcttcttca aggacgacgg taactacaag actagagctg aggttaagtt cgagggtgac
     3781 acattggtta acagaatcga gttgaagggt atcgacttca aagaggacgg aaacatcttg
     3841 ggtcacaagt tggagtacaa ctacaactcc cacaacgttt acatcatggc tgacaagcag
     3901 aagaacggta tcaaggttaa cttcaagatc agacacaaca tcgaggacgg ttccgttcaa
     3961 ttggctgacc actaccaaca gaacactcca attggtgacg gtccagtttt gttgccagac
     4021 aaccactact tgtccactca atccgctttg tccaaggacc caaacgagaa gagagatcac
     4081 atggttttgt tggagttcgt tactgctgct ggtatcactt tgggtatgga cgagttgtac
     4141 aagtaactcg agccttagac atgactgttc ctcagttcaa gttgggcact tacgagaaga
     4201 ccggtcttgc tagattctaa tcaagaggat gtcagaatgc catttgcctg agagatgcag
     4261 gcttcatttt tgatactttt ttatttgtaa cctatatagt ataggatttt ttttgtcatt
     4321 ttgtttcttc tcgtacgagc ttgctcctga tcagcctatc tcgcagctga tgaatatctt
     4381 gtggtagggg tttgggaaaa tcattcgagt ttgatgtttt tcttggtatt tcccactcct
     4441 cttcagagta cagaagatta agtgagacgt tcgtttgtgc aagcttagat ccccgggtac
     4501 cgagctcgaa ttcgccgtac cacttcaaaa cacccaagca cagcatacta aatttcccct
     4561 ctttcttcct ctagggtgtc gttaattacc cgtactaaag gtttggaaaa gaaaaaagag
     4621 accgcctcgt ttctttttct tcgtcgaaaa aggcaataaa aatttttatc acgtttcttt
     4681 ttcttgaaaa tttttttttt tgattttttt ctctttcgat gacctcccat tgatatttaa
     4741 gttaataaac ggtcttcaat ttctcaagtt tcagtttcat ttttcttgtt ctattacaac
     4801 tttttttact tcttgctcat tagaaagaaa gcatagcaat ctaatctaag ttttaattac
     4861 aaaatgaaga agccagagtt gactgctact tccgttgaga agttcttgat cgagaagttc
     4921 gactccgttt ccgacttgat gcaattgtct gaaggtgaag agtccagagc tttctccttt
     4981 gacgttggtg gtagaggtta cgttttgaga gttaattcct gtgctgacgg tttctacaag
     5041 gacagatacg tttacagaca cttcgcttcc gctgctttgc caattccaga ggttttggac
     5101 attggtgagt tctccgagtc cttgacttac tgtatctcca gaagagctca gggagttaca
     5161 ttgcaagact tgccagaaac tgagttgcca gctgttttgc aaccagttgc tgaagctatg
     5221 gacgctattg ctgctgctga cttgtctcaa acttctggtt ttggtccatt cggtccacag
     5281 ggtattggtc agtacactac ttggagagac ttcatctgtg ctatcgctga cccacatgtt
     5341 taccactggc aaactgttat ggacgatact gtttccgctt ccgttgctca ggctttggac
     5401 gaattgatgt tgtgggctga agattgtcct gaggttagac atttggttca cgctgacttc
     5461 ggttccaaca acgttttgac tgacaacggt agaatcactg ctgttatcga ctggtccgaa
     5521 gctatgttcg gtgactctca atacgaggtt gctaacatct tcttttggag accatggttg
     5581 gcttgtatgg aacagcagac tagatacttc gagagaagac acccagaatt ggctggttcc
     5641 ccaagattga gagcttacat gttgagaatc ggtttggacc agttgtacca gtccttggtt
     5701 gatggtaact tcgatgatgc tgcttgggct caaggtagat gtgacgctat cgttagatcc
     5761 ggtgctggta ctgttggtag aactcagatt gctagaagat ccgctgctgt ttggactgac
     5821 ggttgtgttg aagttttggc tgactccggt aacagaagac catccactag accaagagct
     5881 aaagaggttg gtagagttta agtattaaga acgttattta tatttcaaat ttttcttttt
     5941 tttctctaca gacgctgtac gcatgtaaca ttatactgaa aaccttgctt gagaaggttt
     6001 tgggacgctc gaagatccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6061 tgcgtattgg gcgctcttcc gcttcctcgg aattcactgg ccgtcgtttt acaacgtcgt
     6121 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     6181 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     6241 aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     6301 cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     6361 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     6421 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     6481 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     6541 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     6601 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     6661 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     6721 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     6781 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     6841 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     6901 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     6961 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     7021 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     7081 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     7141 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     7201 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     7261 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     7321 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     7381 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     7441 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     7501 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     7561 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     7621 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     7681 cactgagcgt cagaccccgt agaaaagatc aaaggatctt c
//
