LOCUS       sequence-388682-        6723 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMUT2 based vector for sensing lactic acid in E. coli Nissle 1917.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6723)
  AUTHORS   Vaaben TH, Vazquez-Uribe R, Sommer MOA
  TITLE     Characterization of Eight Bacterial Biosensors for Microbial 
            Diagnostic and Therapeutic Applications.
  JOURNAL   ACS Synth Biol. 2022 Dec 16;11(12):4184-4192. doi: 
            10.1021/acssynbio.2c00491. Epub 2022 Nov 30.
  PUBMED    36449712
REFERENCE   2  (bases 1 to 6723)
  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"; doi: 
            "10.1021/acssynbio.2c00491"; journalName: "ACS Synth Biol"; date: 
            "2022-12-16- 16"; volume: "11"; issue: "12"; pages: "4184-4192"
COMMENT     Sequence Label: pMUT2-pLac-mCherry
FEATURES             Location/Qualifiers
     source          1..6723
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     79..98
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     321..338
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(960..1754)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(1755..1859)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2160..2182
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2174..2191
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2175..2191
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2202..2221
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2202..2220
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2225..2236
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      complement(2272..2343)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    3155..3171
                     /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)."
     promoter        complement(join(3179..3185,3186..3201,3202..3207))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     promoter        3305..3339
                     /label=BBa_J23117
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.06"
     RBS             3386..3397
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
       61 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
      121 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
      181 cgcctcccgc gcgcccttct tctgggtctt tgtcttttgc tcacatgttc tttccggttt
      241 tatcccccga ttctgtggat aaccgtatta ccgcctttga gtgagctgac accgctcgcc
      301 gcagtcgaac gaccgagcgt agcgagtcag tgagcgagga agcggaagag cgccttatgt
      361 gacattttct ccttacgctc ctctatgccg ttctgcatcc tgtccggatg cgttatatcc
      421 cggtaagatt ttccgcttca aagcgtgtct gtatgctgtt ctggagttct tctgcgagtt
      481 cgtgcagttt ctcacacatg gcggcctgtt cgtcggcatt gagtgcgtcc agtttttcga
      541 gcagcgtcag gctctgactt tttatgaatc ccgccatgtt gagtacggct tgctgctgct
      601 tattcatctt ttcgttttct ccgttctgtc tgtcatctgc gttgtgtgat tatatcgcgt
      661 accacttttc gactgttttg ctgccgctat tctgccgctt ggctttttga cgggcatttc
      721 tgtcagacaa cactgtcact gccaaaaaac tgccgtgcct ttgtcggtaa ttcgagcttg
      781 ctgacaggac aggatgtgca attgttatac cgcgcataca tgcacgctat tacaattgcc
      841 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
      901 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttat
      961 tagaaaaatt catccagcag acgataaaac gcaatacgct ggctatccgg tgccgcaatg
     1021 ccatacagca ccagaaaacg atccgcccat tcgccgccca gttcttccgc aatatcacgg
     1081 gtggccagcg caatatcctg ataacgatcc gccacgccca gacggccgca atcaataaag
     1141 ccgctaaaac ggccattttc caccataatg ttcggcaggc acgcatcacc atgggtcacc
     1201 accagatctt cgccatccgg catgctcgct ttcagacgcg caaacagctc tgccggtgcc
     1261 aggccctgat gttcttcatc cagatcatcc tgatccacca ggcccgcttc catacgggta
     1321 cgcgcacgtt caatacgatg tttcgcctga tgatcaaacg gacaggtcgc cgggtccagg
     1381 gtatgcagac gacgcatggc atccgccata atgctcactt tttctgccgg cgccagatgg
     1441 ctagacagca gatcctgacc cggcacttcg cccagcagca gccaatcacg gcccgcttcg
     1501 gtcaccacat ccagcaccgc cgcacacgga acaccggtgg tggccagcca gctcagacgc
     1561 gccgcttcat cctgcagctc gttcagcgca ccgctcagat cggttttcac aaacagcacc
     1621 ggacgaccct gcgcgctcag acgaaacacc gccgcatcag agcagccaat ggtctgctgc
     1681 gcccaatcat agccaaacag acgttccacc cacgctgccg ggctacccgc atgcaggcca
     1741 tcctgttcaa tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     1801 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     1861 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
     1921 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     1981 ataaatcaaa agaatagacc gagatagggt tgagtggccg ctacagggcg ctcccattcg
     2041 ccattcaggc tgcgcaactg ttgggaaggg cgtttcggtg cgggcctctt cgctattacg
     2101 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     2161 ccagtcacga cgttgtaaaa cgacggccag tgagcgcgac gtaatacgac tcactatagg
     2221 gcgaattgaa ggaaggccgt caaggccacg tgtcttgtcc gcggtaccta cgagagcgtt
     2281 caccgacaaa caacagataa aacgaaaggc ccagtctttc gactgagcct ttcgttttat
     2341 ttgatgcctg gtcatgcgtt tttctccctc gaatgctcat tatggtcacc aggcaggcgg
     2401 gtaatacgtg catggcgagc ctgatcttca tcgaatcgtt tcatggtggt gtgaacaaag
     2461 ctaaggtgcg ccatcattgc tttacgcgcc ccgtcagcat caccggcaaa aatggcgtca
     2521 atgaccgcct gatgttgttc ggtcagctgt gaaaaaaccg ggggtaccag gtacatccgc
     2581 tggcggctat gcttcacaga ggattgcagg acatcgaaga aaccgcgcat ggtttgcagc
     2641 aacacaatgt tatgtgaagc ttcggcaatc gccagatgaa aacgaacgtc cgcttgtgag
     2701 gcgagatccg ggtcttcact tagcgttgct tcaaagcaaa gctgaatctt ttctttttct
     2761 ccaggtgtgg cgcgcattgc tgcatgccat gcggtgctgg cttcaatggc gtatcgggct
     2821 tccagaatat cgaaactgta atccggatca tcggccatca gcggtttcag cggctggacg
     2881 atgttttgct ccgaccatgt gtcatgacgc cagcgaataa acgtcccgcc gccacgtcga
     2941 ctgagcagca caccttcgct taccagtttc gccagcgcct cgcgcagtga atttctcgac
     3001 acgccgagtt gcatcgccag ttgacgttca gcgggcaact tcatgcccgc ttccaggttt
     3061 ttttcatcaa tcagcgcccg cacacgatcg gcaacctcgt ctgacaggcg tctgggtaaa
     3121 acaatcatct agtatttctc tcttttgtgt gaaattgtta tccgctcaca attccacaca
     3181 ttatacgagc cgatgattaa ttgtcaagcg ccccagctgg caattccgac gtcaattggc
     3241 cctaccaatt cttcgcttat ctgacctctg gttcacaatt tcccaattaa aactcacatc
     3301 aatgttgaca gctagctcag tcctagggat tgtgctagct cattatccct acacaacaca
     3361 attggcagtg ccacttgaat tcattaaaga ggagaaaggt acctcaggag aattacaatg
     3421 gtttccaagg gcgaggagga taacatggct atcattaaag agttcatgcg cttcaaagtt
     3481 cacatggagg gttctgttaa cggtcacgag ttcgagatcg aaggcgaagg cgagggccgt
     3541 ccgtatgaag gcacccagac cgccaaactg aaagtgacta aaggcggccc gctgcctttt
     3601 gcgtgggaca tcctgagccc gcaatttatg tacggttcta aagcgtatgt taaacaccca
     3661 gcggatatcc cggactatct gaagctgtct tttccggaag gtttcaagtg ggaacgcgta
     3721 atgaattttg aagatggtgg tgtcgtgacc gtcactcagg actcctccct gcaggatggc
     3781 gagttcatct ataaagttaa actgcgtggt actaattttc catctgatgg cccggtgatg
     3841 cagaaaaaga cgatgggttg ggaggcgtct agcgaacgca tgtatccgga agatggtgcg
     3901 ctgaaaggcg aaattaaaca gcgcctgaaa ctgaaagatg gcggccatta tgacgctgaa
     3961 gtgaaaacca cgtacaaagc caagaaacct gtgcagctgc ctggcgcgta caatgtgaat
     4021 attaaactgg acatcacctc tcataatgaa gattatacga tcgtagagca atatgagcgc
     4081 gcggagggtc gtcattctac cggtggcatg gatgagctgt acaaataaaa aggggctacc
     4141 ggcgaaccag cagccccttt ataaaggcgc ttcagtagtc agaccagcat cagtcctgaa
     4201 aaggcgggcc tgcgcccgcc tccaggttgc tacttaccgg attcgtaagc catgaaagcc
     4261 gccacctccc tgtgtccgtc tctgtaacga atctcgcaca gcgattttcg tgtcagataa
     4321 gtgaatatca acagtgtgag acacacgatc aacacacacc agacaaggga acttcgtggt
     4381 agtttcatgg ccttcttctc cttgcgcaaa gcgcggtaag aggctatcct gatgtggact
     4441 agacataggg atgcctcgtg gtggttaatg aaaattaact tactacgggg ctatcttctt
     4501 tctgccacac aacacggcaa caaaccacct tcacgtcatg aggcagaaag cctcaagcgc
     4561 cgggcacatc atagcccata tacctgcacg ctgaccacac tcactttccc tgaaaataat
     4621 ccgctcattc agaccgttca cgggaaatcc gtgtgattgt tgccgcatca cgctgcctcc
     4681 cggagtttgt tcggcataag tcacaagaga cactcaatgg caaaggaaga ccgtgccctg
     4741 ttatccaggc ttatggcaga ttagcttccc ggagagaaaa ctgtcgaaaa ctgacggtat
     4801 gaacaccgta agctcccaaa gtgatcacca ttcgctttca tgcatagcta tgcagcgagc
     4861 tgaaaacgat cctgacgcat ccttcctgtt tttccggggt aaaacatctc tttttgcggt
     4921 gtctgcgtca gaatcgcgtt cagcgcgttt cagtggtgcg tacaattaag ggattatggt
     4981 aaatatatga gctatgcgat aactttaact gtgaagcgat gaacccatta caggcaaagc
     5041 caattactcc tgacagtggt ttagccagaa gcagggctac caagaccaat gcaataagta
     5101 atatatcgtt ttgctatcgt gcatcgctcg cgctcagttc cattgtgctt ttttaagctg
     5161 tcgtttttct tacggtatat accggttttt tatggcgtgg tttcttaact tgttcagcta
     5221 ctgatggacc cattgtatct aggtagtcaa ctagctttgt tagatcataa atattgcgac
     5281 caccatatcg gcgatcactc ttcgatgttg gtttcttgtc caagagatta gctttttcaa
     5341 gatcattgat agctctctga acagtccgta cagaaacccc catacgtatg gctagacttt
     5401 ccattgacgg atgcggccac tcttgcaaac tccaccagtg aacgatcagg ttaagtagtg
     5461 tgttaaaggc cactgaagtt agctttttct cgttttgtat aaaaaacaat acggtaggca
     5521 ctgctgtcca gccaagagac aaaccgccag ctttccattt attcttaacg gagtaagtca
     5581 ttgattttcc taagccccaa aatatttaaa gtatatatta tatggtatat tcatatgaat
     5641 agggtgacac tggcgccatt attgtgcaac caaaaaagac tactctgaaa acgaggaaaa
     5701 gattttttcc tgcctgaatt agatacggag ttagcgatat gaaaaccgaa caacgtcatg
     5761 atcttgttaa agatattgag gtttttggcg tatccttgtc tctgttgatt tccagagcga
     5821 atgagaagtc tgttacaatg ccatctggtc taagtcggga gcagagaaga gcatgggcag
     5881 cggagcaggc gcgcaaaatc cacaattgaa tattgtctca ttctctgaga ccttcaacct
     5941 ttattacaca tccagatatt ctgcaaaaac actcgataaa atcgatgatt tcattgagca
     6001 ttttgaaaaa tacaatctct ttggcgatcc tttaaaagga tatccagctt ggactggcaa
     6061 agtatcgcca tcgtggaaag tgcctgatca ttacgaaaac aaagaagcta ttgagaagta
     6121 tgctagagct aacaaattat ggcatgctca tttaggcgat ccggttttta aagatacgtt
     6181 tcatgggaaa tacaaggttt ctgactgggt tattcatttc cagcggctga caccgaacca
     6241 tataaagctt ttagagcttg gataccatga cccaatgaag ctacctcaaa actttgaatg
     6301 atcgagcggc aggctaaatg aaatcttgag attcattcag tctcgtcgta atctcactat
     6361 tgtaaaaacg aaaaaaccac cctggcaggt ggtttttcga aggttagtta atcctggcag
     6421 attctctaac cgtggtaaca gtcttgtgcg agacatgtca ccaaatactg tcctttcagt
     6481 gtagcctcag ttaggccgcc acttcaagaa ctctcgttac atctctcgca catcctgctt
     6541 accagtggcc gttgccagtg gcgttaagtc gtgtcttacc gggttggact caagacgata
     6601 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6661 ggagcgaacg acctacaccg aaccgagata ccaacagcgt gagctatgag aaagcgccac
     6721 gct
//
