LOCUS       sequence-346802-        5404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lactate biosensor in anaerobic conditions.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5404)
  AUTHORS   Zuniga A, Camacho M, Chang HJ, Fristot E, Mayonove P, Hani EH, 
            Bonnet J
  TITLE     Engineered l-Lactate Responding Promoter System Operating in 
            Glucose-Rich and Anoxic Environments.
  JOURNAL   ACS Synth Biol. 2021 Dec 1. doi: 10.1021/acssynbio.1c00456.
  PUBMED    34851606
REFERENCE   2  (bases 1 to 5404)
  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: "ACS Synth 
            Biol. 2021 Dec 1. doi: 10.1021/acssynbio.1c00456."
COMMENT     Sequence Label: ALPaGA
FEATURES             Location/Qualifiers
     source          1..5404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1196..1207
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(1214..1216,1217..1930)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2025..2096
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2112..2139
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2297..3112)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3020..3039
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3104..3123
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(3470..4420)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      4468..4690
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5182..5276)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 ctcggatacc cttactctgt tgaaaacgaa tagataggtt tcatgcgttt ttctccctcg
       61 aatgctcatt atgctcaccg ggcaggcggg taatccgtgc gtggcgagcc tgatcttcat
      121 cgaatcgttt catggtggtg tgaacaaaac taaggtgcgc catcattgct ttacgcgccc
      181 cgtcagcatc accggcaaaa atggcgtcaa tgacagcctg atgttgttcg gtcagttgtg
      241 aaaaaaccgg tggcaccaga tacatccgct gacggctatg cttcactgag gattgcagga
      301 catcgaagaa accgcgcatg gtttgcagca gcacgatgtt atgtgaggct tcggcaatcg
      361 ccagatgaaa acgaacgtcc gcttgtgagg cgatatccgg gtcttcactt agcgttgctt
      421 caaagcaaag ctgaatcttt tctttgtcgc caggtgtggc gcgcattgcc gcatgccatg
      481 cggtgctggc ttcaatggcg tagcgggctt ccagaatatc gaaactgtaa tccggatcat
      541 cggccatcag tgtttttagc ggctggacga tgttttgctc cgaccatgtg tcatgacgcc
      601 agcgaataaa cgtcccgccg ccgcgtcgac tgagcagcac gccttcactc accagttttg
      661 ccagcgcctc gcgcagtgaa ttacgtgata cgccgagttg catcgccagt tggcgctcag
      721 cgggcaactt catgcccgct tccaggtttt tttcatcaat cagcgcccgc acacgatcgg
      781 caacctcgtc tgacaggcgt ctgggtaaaa caatcatact agtgggtcct gtctttctct
      841 agatccacac attatacgag ccgatgatta agaggcaaag attactactg ataaccactg
      901 ttgattggga tacccgtaag agaatataaa aagccagatt attaatccgg cttttttatt
      961 atttctttac cagacatctc cccccacaag aattggccct accaattctt cgcttaaaag
     1021 gtcccgtctg aacttacgtc gcttttaact attcttaaac ttgacaatta atcatccggc
     1081 tcttagtgtt tgtggagtca ttatccctac acaacacaat tggcagtgcc acttttacac
     1141 aacgtgtgac aaggagatga gcaacagact cattacacga tgtgcgtgga ctccaaaaga
     1201 ggagaaatac tagatgtcaa aaggagaaga actttttaca ggtgtagtac ctatcttggt
     1261 tgaattggat ggtgatgtta acggtcacaa attttctgta cgtggtgaag gtgaaggtga
     1321 tgcaactaac ggtaaattga cacttaaatt catttgtaca actggaaaac ttcctgttcc
     1381 ttggcctact cttgttacaa cattgacata tggagtacaa tgtttttcac gttatcctga
     1441 tcatatgaaa cgtcacgatt tttttaaatc tgctatgcca gaaggttatg tacaagaacg
     1501 tacaatttca tttaaagatg acggaacata taaaacacgt gctgaagtaa aattcgaagg
     1561 tgacactctt gttaatcgta tcgaattgaa aggaatcgat ttcaaagaag atggtaacat
     1621 tttgggacac aaacttgaat acaacttcaa ctctcataat gtttatatca cagctgacaa
     1681 acaaaaaaac ggtattaaag ctaattttaa aattcgtcac aatgttgaag atggatctgt
     1741 tcaattggct gatcattatc aacaaaatac accaatcgga gacggaccag tattgcttcc
     1801 agataaccac tacctttcta ctcaatcagt tctttcaaaa gatcctaacg aaaaacgtga
     1861 ccatatggta cttcttgaat ttgttacagc agcaggtatc actcacggta tggacgaact
     1921 ttataaataa actttatctg agaatagtca atcttcggaa atcccaggtg gcatgctaaa
     1981 agtctcgtaa agcgttctat caataacccg ttggtgccag gcatcaaata aaacgaaagg
     2041 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctctact
     2101 agagtcacac tggctcacct tcgggtgggc ctttctgcgt ttataccgtc tcagaatcgg
     2161 ccgtgaacaa taaaatagtt tcggtattat tgaccacttc cgagtagaat cgtgcttcag
     2221 taagagctca ctcaaaggcg gtaatcagat aaaaaaaatc cttagctttc gctaaggatg
     2281 atttctgcta gtattattag aaaaactcat cgagcatcaa atgaaactgc aatttattca
     2341 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
     2401 caccgaggca gttccaaaga atggcaaggt cctggtaacg gtctgcgatt ccgacccgtc
     2461 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
     2521 caccatgagt gacgactgaa tccggtgaga atggcaagag cttgtgcatt tctttccaga
     2581 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
     2641 tattcatgcg tgattgcgcc tgagcaagac gaaatacacg atcgctgtta aaaggacaat
     2701 tacaaacagg aatcgaatgt aaccggcgca ggaacacggc cagcgcatca acaatatttt
     2761 cacctgaatc aggatattct tctaatacct ggaaggctgt tttcccagga atcgcggtgg
     2821 tgagtaacca cgcatcatca ggagtacgga taaaatgctt gatggtcggg agaggcataa
     2881 actccgtcag ccagttgaga cggaccatct catctgtaac atcattggca acgctacctt
     2941 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaagcga tagattgtcg
     3001 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcgtccatgt
     3061 tggagtttaa gcgcggacgg gagcaagacg tttcccgttg aatatggctc ataacacccc
     3121 ttgtattact gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt
     3181 gtgcaatgta acatcagaga ttttgagaca caacgtggct ttgttgaata aatcgaactt
     3241 ttgctgagtt gaaggatcag ctctagtagt tacattgtcg atctgttcat ggtgaacagc
     3301 tttgaatgca ccaaaaactc gtaaaagctc tgatgtatct atctttttta caccgttttc
     3361 atctgtgcat atggacagtt ttccctttga tatgtaacgg tgaacagttg ttctactttt
     3421 gtttgttagt cttgatgctt cactgataga tacaagagcc ataagaacct cagatccttc
     3481 cgtatttagc cagtatgttc tctagtgtgg ttcgttgttt ttgcgtgagc catgagaacg
     3541 aaccattgag atcatactta ctttgcatgt cactcaaaaa ttttgcctca aaactggtga
     3601 gctgaatttt tgcagttaaa gcatcgtgta gtgtttttct tagtccgtta tgtaggtagg
     3661 aatctgatgt aatggttgtt ggtattttgt caccattcat ttttatctgg ttgttctcaa
     3721 gttcggttac gagatccatt tgtctatcta gttcaacttg gaaaatcaac gtatcagtcg
     3781 ggcggcctcg cttatcaacc accaatttca tattgctgta agtgtttaaa tctttactta
     3841 ttggtttcaa aacccattgg ttaagccttt taaactcatg gtagttattt tcaagcatta
     3901 acatgaactt aaattcatca aggctaatct ctatatttgc cttgtgagtt ttcttttgtg
     3961 ttagttcttt taataaccac tcataaatcc tcatagagta tttgttttca aaagacttaa
     4021 catgttccag attatatttt atgaattttt ttaactggaa aagataaggc aatatctctt
     4081 cactaaaaac taattctaat ttttcgcttg agaacttggc atagtttgtc cactggaaaa
     4141 tctcaaagcc tttaaccaaa ggattcctga tttccacagt tctcgtcatc agctctctgg
     4201 ttgctttagc taatacacca taagcatttt ccctactgat gttcatcatc tgagcgtatt
     4261 ggttataagt gaacgatacc gtccgttctt tccttgtagg gttttcaatc gtggggttga
     4321 gtagtgccac acagcataaa attagcttgg tttcatgctc cgttaagtca tagcgactaa
     4381 tcgctagttc atttgctttg aaaacaacta attcagacat acatctcaat tggtctaggt
     4441 gattttaatc actataccaa ttgagatggg ctagtcaatg ataattacat gtccttttcc
     4501 tttgagttgt gggtatctgt aaattctgct agacctttgc tggaaaactt gtaaattctg
     4561 ctagaccctc tgtaaattcc gctagacctt tgtgtgtttt ttttgtttat attcaagtgg
     4621 ttataattta tagaataaag aaagaataaa aaaagataaa aagaatagat cccagccctg
     4681 tgtataactc actactttag tcagttccgc agtattacaa aaggatgtcg caaacgctgt
     4741 ttgctcctct acaaaacaga ccttaaaacc ctaaaggctt aagtagcacc ctcgcaagct
     4801 cgggcaaatc gctgaatatt ccttttgtct ccgaccatca ggcacctgag tcgctgtctt
     4861 tttcgtgaca ttcagttcgc tgcgctcacg gctctggcag tgaatggggg taaatggcac
     4921 tacaggcgcc ttttatggat tcatgcaagg aaactaccca taatacaaga aaagcccgtc
     4981 acgggcttct cagggcgttt tatggcgggt ctgctatgtg gtgctatctg actttttgct
     5041 gttcagcagt tcctgccctc tgattttcca gtctgaccac ttcggattat cccgtgacag
     5101 gtcattcaga ctggctaatg cacccagtaa ggcagcggta tcatcaacag gcttacccgt
     5161 cttactgtcc ctagtgcttg gattctcacc aataaaaaac gcccggcggc aaccgagcgt
     5221 tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca ggagtccaag
     5281 cgagctcgta aacttggtct gacagctcta gctccggcaa aaaaacgggc aaggtgtcac
     5341 caccctgccc tttttcttta aaaccgaaaa gattacttcg cgtttgccac ctgacgtcta
     5401 agaa
//
