LOCUS       sequence-408886-        5834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Biosensor for extracellular L-lactate.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5834)
  AUTHORS   Nasu Y, Aggarwal A, Le GNT, Vo CT, Kambe Y, Wang X, Beinlich FRM, 
            Lee AB, Ram TR, Wang F, Gorzo KA, Kamijo Y, Boisvert M, Nishinami S,
            Kawamura G, Ozawa T, Toda H, Gordon GR, Ge S, Hirase H, Nedergaard 
            M, Paquet ME, Drobizhev M, Podgorski K, Campbell RE
  TITLE     Lactate biosensors for spectrally and spatially multiplexed 
            fluorescence imaging.
  JOURNAL   Nat Commun. 2023 Oct 27;14(1):6598. doi: 10.1038/s41467-023-42230-5.
  PUBMED    37891202
REFERENCE   2  (bases 1 to 5834)
  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: "2023-10-27"; volume: "14(1):6598. doi"; pages: " 
            10.1038/s41467-023-42230-5"
COMMENT     Sequence Label: pBAD-eLACCO2.1
FEATURES             Location/Qualifiers
     source          1..5834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             331..348
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             352..384
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(388..396,397..411)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(1345..1366)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1606..1625)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(2235..2252)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2405..2491
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2583..2610
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2629..2720
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2721..2789,2790..3581)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2939..2958)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3752..4340
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4241..4260
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4494..4511
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4526..4666
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4592..4611)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4752..4774
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4930..5808)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLS*REDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aattaaccat ggggggttct catcatcatc atcatcatgg tatggctagc
      361 atgactggtg gacagcaaat gggtcgggat ctgtacgacg atgacgataa ggatccgagc
      421 tcgagaatgt tctctccctt ggcagtggcc caggctcgac gctataggtg gcgcattcaa
      481 accgcgtggg atgccggaac agtaggctac agtctgttcc aaaagtttac ggagcgggta
      541 aaggaactga cggatggaca gcttgaggtc cagccttttc cagcgagtgc agtcgtaggc
      601 acgtttgata tgttcgacgc ggtcaagact ggagttttgg acggaatgaa tcctttcacg
      661 atctattggg cagggaggat gccggttaca gctttcctct ccagctacgc cctcgggttg
      721 gaccgacctg accagtggga aacatggttt tactccctgg gtggcctcga caacgcccga
      781 cgagcctttg ccgagcaagg cctcttctac gtgggaccag ttcagcacga cctcaataca
      841 attcattcaa agaagccgat ccggagattt gaggacttca aaggtgtcaa gctccgagtg
      901 ccgggcggca tgattgcgga agtttttgcg gccgcaggag cttccaccgt cctcttgcct
      961 gggggggagg tcaaccccgc tctggaaaga ggggtaatag attggagcca caacgtccat
     1021 atcatggccg acaagcagag gaacggcatc aaggcgaact tcgagatccg ccacagcacc
     1081 gaggacggcg gcgtgcagct cgcctaccac taccagcaga acacccccat cggcgacggc
     1141 cccgtgctgc tgcccgacaa ccactacctg agcacccaga ccaaacttac gaaagacccc
     1201 aacgagaagc gcgatcacat ggtcctgctg gagtacgtga ccgccgccgg gatcactcgc
     1261 ggcatggacg agctgtacaa gggcggtacc ggagggagaa tggtgagcaa gggcgaggag
     1321 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1381 ttcagcgtgt ccggcgaggg tgagggcgat gccacctccg gcaagctgtc cctgaagttc
     1441 atcagcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     1501 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcg
     1561 gccatgcccg aaggctacat ccaggagcgc accatcttct tcaaggacga cggcaactac
     1621 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1681 ggcatcgact tcaaggagga cggcaacatc ctggggcata agctggagta cagctttaac
     1741 gatggtggtg cggctgattt tgttgggcct gcagtaaact ataacttggg gttccatcag
     1801 gtggcaaagt acatcataat ggggcctcca gagacgcctg ccattcatca acctgtagac
     1861 ctcatggact tcaccataaa ccttaatcgg tggcgcagcc ttccaaagcc cctgcaagaa
     1921 agattcatcg cggcggttca cgagtactct tggattcact acgcaggcat tcagaaggct
     1981 aatttggagg cgtggccgaa atatcggcag gcaggtgttg aggtcatccg gctgagcaac
     2041 gaggatgtta gaaaattcag gcgactcgct attcccattt ggtttaagtg ggctaagatg
     2101 gataactact cacgagaggc attcgcgtct caactggagt atatgaaagg tataggttac
     2161 gttacggacg aggaattgaa gggcttgggc ctgtgaaagc ttggctgttt tggcggatga
     2221 gagaagattt tcagcctgat acagattaaa tcagaacgca gaagcggtct gataaaacag
     2281 aatttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg
     2341 aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccaa
     2401 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     2461 gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa
     2521 gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa
     2581 gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttt tgtttatttt
     2641 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     2701 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     2761 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     2821 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     2881 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     2941 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
     3001 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     3061 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     3121 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     3181 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     3241 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     3301 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     3361 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     3421 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     3481 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     3541 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     3601 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     3661 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     3721 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3781 gctgcttgca aacaaaacaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3841 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     3901 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3961 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4021 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4081 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4141 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4201 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4261 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4321 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4381 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     4441 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     4501 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     4561 gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     4621 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
     4681 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     4741 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     4801 gaggcagcag atcaattcgc gcgcgaaggc gaagcggcat gcataatgtg cctgtcaaat
     4861 ggacgaagca gggattctgc aaaccctatg ctactccgtc aagccgtcaa ttgtctgatt
     4921 cgttaccaat tatgacaact tgacggctac atcattcact ttttcttcac aaccggcacg
     4981 gaactcgctc gggctggccc cggtgcattt tttaaatacc cgcgagaaat agagttgatc
     5041 gtcaaaacca acattgcgac cgacggtggc gataggcatc cgggtggtgc tcaaaagcag
     5101 cttcgcctgg ctgatacgtt ggtcctcgcg ctagcttaag acgctaatcc ctaactgctg
     5161 gcggaaaaga tgtgacagac gcgacggcga caagcaaaca tgctgtgcga cgctggcgat
     5221 atcaaaattg ctgtctgcca ggtgatcgct gatgtactga caagcctcgc gtacccgatt
     5281 atccatcggt ggatggagcg actcgttaat cgcttccatg cgccgcagta acaattgctc
     5341 aagcagattt atcgccagca gctccgaata gcgcccttcc ccttgcccgg cgttaatgat
     5401 ttgcccaaac aggtcgctga aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt
     5461 attggcaaat attgacggcc agttaagcca ttcatgccag taggcgcgcg gacgaaagta
     5521 aacccactgg tgataccatt cgcgagcctc cggatgacga ccgtagtgat gaatctctcc
     5581 tggcgggaac agcaaaatat cacccggtcg gcaaacaaat tctcgtccct gatttttcac
     5641 caccccctga ccgcgaatgg tgagattgag aatataacct ttcattccca gcggtcggtc
     5701 gataaaaaaa tcgagataac cgttggcctc aatcggcgtt aaacccgcca ccagatgggc
     5761 attaaacgag tatcccggca gcaggggatc attttgcgct tcagccatac ttttcatact
     5821 cccgccattc agag
//
