LOCUS       sequence-457086-        6107 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of LuxTA mutant with strong promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6107)
  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 6107)
  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: pMT1035
FEATURES             Location/Qualifiers
     source          1..6107
                     /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"
     promoter        2256..2732
                     /label=GAP promoter
                     /note="promoter for Pichia pastoris 
                     glyceraldehyde-3-phosphate dehydrogenase"
     CDS             2885..2905
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      3731..3977
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             complement(4626..5420)
                     /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(4737..4756)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5347..5366
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(5713..5730)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5713..5729)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5722..5744)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5938..5957)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6057..6079
                     /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 atcctttttt gtagaaatgt cttggtgtcc
     2281 tcgtccaatc aggtagccat ctctgaaata tctggctccg ttgcaactcc gaacgacctg
     2341 ctggcaacgt aaaattctcc ggggtaaaac ttaaatgtgg agtaatggaa ccagaaacgt
     2401 ctcttccctt ctctctcctt ccaccgcccg ttaccgtccc taggaaattt tactctgctg
     2461 gagagcttct tctacggccc ccttgcagca atgctcttcc cagcattacg ttgcgggtaa
     2521 aacggaggtc gtgtacccga cctagcagcc cagggatgga aaagtcccgg ccgtcgctgg
     2581 caataatagc gggcggacgc atgtcatgag attattggaa accaccagaa tcgaatataa
     2641 aaggcgaaca cctttcccaa ttttggtttc tcctgaccca aagactttaa atttaattta
     2701 tttgtcccta tttcaatcaa ttgaacaact atcaaaacac aactagtgcg tcgacatgcc
     2761 agccgatgct ttggatgatt tcgatttgga tatgttgcct gctgatgcat tggacgattt
     2821 tgacttagac atgttaccag cagacgcatt ggatgacttc gaccttgata tgctaccagg
     2881 tccaccaaaa aagaagagaa aggttgaaaa catcaacgct gatgacacct acagaatcat
     2941 caacaagatt aaggcttgca ggtctaacaa cgatattaac cagtgtttgt ccgacatgac
     3001 caagatggtt cattgcgaat attacttgtt ggccatcatc tacccacact ctatggttaa
     3061 gtccgacatt tccattttgg acaactaccc aaaaaaatgg cgtcagtatt acgatgatgc
     3121 caacttgatt aagtacgacc caatcgttga ttactccaac tctaatcatt ctcccatcaa
     3181 ctggaacatc ttcgaaaaca atgccgtcaa caaaaagtcc ccaaacgtta tcaaagaagc
     3241 taagacctat ggtttgatta ccggtttttc tttcccaatt cataccgcta acaatggttt
     3301 cggtatgttg tcttttgccc actctgaaaa ggataactac atcgactcat tattcttgca
     3361 tgcctgcatg aacattccat tgatagttcc atctctggtc gataactata ggaagattaa
     3421 cattgccaac aacaagtcca acaacgattt gaccaaaagg gaaaaagaat gtttggcttg
     3481 ggcttgcgaa ggtaaatctt ctagggatat ttccaagatt ttgggttgct ctgaaagaac
     3541 cgttactttc catttgacta acgcccaaat gaagttgaac actaccaaca gatgccagtc
     3601 tatttctaag gctattttga ccggtgctat tgattgtcca tactttaaga attaacctcg
     3661 agccttagac atgactgttc ctcagttcaa gttgggcact tacgagaaga ccggtcttgc
     3721 tagattctaa tcaagaggat gtcagaatgc catttgcctg agagatgcag gcttcatttt
     3781 tgatactttt ttatttgtaa cctatatagt ataggatttt ttttgtcatt ttgtttcttc
     3841 tcgtacgagc ttgctcctga tcagcctatc tcgcagctga tgaatatctt gtggtagggg
     3901 tttgggaaaa tcattcgagt ttgatgtttt tcttggtatt tcccactcct cttcagagta
     3961 cagaagatta agtgagacgt tcgtttgtgc aagcttagat cttctagaga caataagaag
     4021 aaaaaaaaag aaaagcggtg ggggagggat tattaaataa ggattatgta accccagggt
     4081 accgttctat acatatttaa ggattattta ggacaatcga tgaaatcggc atcaaactgg
     4141 atgggagtat agtgtccgga taatcggata aatcatcttg cgaggagccg cttggttggt
     4201 tggtgagagg agtgaaatat gtgtctcctc acccaagaat cgcgatatca gcaccctgtg
     4261 ggggacacta ttggcctccc tcccaaacct tcgatgtggt agtgctttat tatattgatt
     4321 acattgatta catagctaaa ccctgcctgg ttgcaagttg agctccgaat tccaatatta
     4381 gtaaaatgcc tgcaagataa cctcggtatg gcgtccgacc ccgcttaatt attttaactc
     4441 ctttccaacg aggacttcgt aatttttgat tagggagttg agaaacgggg ggtcttgata
     4501 cctcctcgat ttcagatccc accccctctc agtcccaagt gggacccccc tcggccgtga
     4561 aatgcgcgca ctttagtttt tttcgcatgt aaacgccggt gtccgtcatc tagagaattc
     4621 cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg
     4681 gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata
     4741 tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg
     4801 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg
     4861 gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct
     4921 ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc
     4981 cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga
     5041 tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca
     5101 aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc
     5161 gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat
     5221 agccgcgctg cctcgtcctg cagttcattc agggcaccgg acaggtcggt cttgacaaaa
     5281 agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc
     5341 tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc
     5401 aatccatctt gttcaatcat atggtttcta tattatcttt gtactaaaga gcaattgata
     5461 atgtgcgaga aaaactggtc cttatatacc gtttgcagca ctccctcccg aactttacga
     5521 aaagtcgtgc gccacctgat tttcatcacg ccaaaaacct acacgtatga ctactccggg
     5581 ccagtgttca ccacgagcta tatagtgtta attaattacc ttattggtta gctctgcatg
     5641 taagggtggt gtgagccggg aattgggtct actctagcgt tcagtaaggt gatataaagc
     5701 tctgtagaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     5761 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     5821 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg
     5881 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac
     5941 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     6001 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     6061 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcga
//
