LOCUS       sequence-457081-        6081 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of PhlTA mutant with weak promoter and 
            terminator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6081)
  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 6081)
  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: pMT1026
FEATURES             Location/Qualifiers
     source          1..6081
                     /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             3498..3518
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(4600..5394)
                     /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(4711..4730)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5321..5340
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(5687..5704)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5687..5703)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5696..5718)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5912..5931)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6031..6053
                     /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 atccgagtca caatctgctt ccacagacga
     2281 gtacaaggac aggcaaaagg aattggaaga agttgctaac ccaataatga gcaagttcta
     2341 tggagctgct ggtggagctc ctggtggagc tcctggtggc ttccctggag gtttccctgg
     2401 cggagctggc gcagctggcg gtgccccagg tggtgctgcc ccaggcggag acagcggacc
     2461 aaccgtggaa gaagtcgatt aagcaattca acggataaat tctggttaat atatataacg
     2521 tgaataggaa attaaggaaa ttttggatct aataatgtgc tgtatgccga catcgggcat
     2581 cgtagattgt atagtatcgc tgacactata ataagccagc caaaacccct aaaccagttg
     2641 ccctccacta attagtgtac tacccaatct tgcctcttcg ggtgtctttt ataaggacag
     2701 attcacaagc tcttgttgcc caatacacac atacacacag agataatagc agtcactagt
     2761 gcgtcgacat ggctagaact ccatccagat catctattgg ttctttgaga tcaccacata
     2821 ctcataaggc tattttgacc tccaccatcg aaatcttgaa agaatgtggt tactccggtt
     2881 tgtccattgg atctgttgct agacgtgctg gtgcttctaa acctactatc tacagatggt
     2941 ggactaacaa ggctgctttg attgctgaag tttacgaaaa cgaatccgaa caggttagaa
     3001 agtttccaga tttgggttcc tttaaggccg atttggattt cttgttgaga aacttgtgga
     3061 aggtttggag agaaactatt tgtggtgaag ccttcagatg cgttattgca gaagctcaat
     3121 tagatccagc tactttgact caattgaagg accaattcat ggaaagacgt agagaaatgc
     3181 caaaaaagtt ggttgaaaac gccatctcta atggtgaatt gccaaaggat acaaaccgtg
     3241 agttgttgtt ggatatgatt ttcggttttt gctggtacag gttgttgact gaacaattga
     3301 ctgttgaaca agacatcgaa gagttcacgt tcttgttaat caatggtgtt tgtccaggta
     3361 ctcaaagagg tccagcagac gctttggatg attttgattt ggatatgttg cctgctgatg
     3421 ccttggatga cttcgactta gacatgttac cagccgatgc attagacgat ttcgaccttg
     3481 atatgttacc tggtccacca aaaaagaaga gaaaggtcta acctcgagcg cattttacca
     3541 catctctact tgttctcacg accccttgca tcgcatatcc tacacatttt aaatcggcat
     3601 tccttccaag catattctta attcctaatt ttacatagtt tttttttttt tttagagaaa
     3661 ataacaacaa tatacataga acttacttac tcaattctac tatatctttt ttttttgtta
     3721 ctattagtca tgtttgcatg actccattac attttcgttt ttttgctagc caattctggt
     3781 cgtcggaaaa acttttcatt gaatctttca aagcgtgtag cagttgcatt tttttaaaaa
     3841 tgaatatgta tgcaaaatat atatatgtat atatatgcct aatatacata ataatagaaa
     3901 aaaaaaacct catacgaaat aggcgcttct cgtatctata ctcaacccgc ccccaatgct
     3961 gaaaattgtt agatcttcta gagacaataa gaagaaaaaa aaagaaaagc ggtgggggag
     4021 ggattattaa ataaggatta tgtaacccca gggtaccgtt ctatacatat ttaaggatta
     4081 tttaggacaa tcgatgaaat cggcatcaaa ctggatggga gtatagtgtc cggataatcg
     4141 gataaatcat cttgcgagga gccgcttggt tggttggtga gaggagtgaa atatgtgtct
     4201 cctcacccaa gaatcgcgat atcagcaccc tgtgggggac actattggcc tccctcccaa
     4261 accttcgatg tggtagtgct ttattatatt gattacattg attacatagc taaaccctgc
     4321 ctggttgcaa gttgagctcc gaattccaat attagtaaaa tgcctgcaag ataacctcgg
     4381 tatggcgtcc gaccccgctt aattatttta actcctttcc aacgaggact tcgtaatttt
     4441 tgattaggga gttgagaaac ggggggtctt gatacctcct cgatttcaga tcccaccccc
     4501 tctcagtccc aagtgggacc cccctcggcc gtgaaatgcg cgcactttag tttttttcgc
     4561 atgtaaacgc cggtgtccgt catctagaga attccccgct cagaagaact cgtcaagaag
     4621 gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg
     4681 gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg
     4741 atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc
     4801 caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg
     4861 catgcgcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc
     4921 cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg
     4981 tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc
     5041 atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc
     5101 cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc
     5161 tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cctgcagttc
     5221 attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag
     5281 ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag
     5341 cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatatggtt
     5401 tctatattat ctttgtacta aagagcaatt gataatgtgc gagaaaaact ggtccttata
     5461 taccgtttgc agcactccct cccgaacttt acgaaaagtc gtgcgccacc tgattttcat
     5521 cacgccaaaa acctacacgt atgactactc cgggccagtg ttcaccacga gctatatagt
     5581 gttaattaat taccttattg gttagctctg catgtaaggg tggtgtgagc cgggaattgg
     5641 gtctactcta gcgttcagta aggtgatata aagctctgta gaattcactg gccgtcgttt
     5701 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     5761 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     5821 tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg
     5881 gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     5941 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     6001 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     6061 cgtcatcacc gaaacgcgcg a
//
