LOCUS       sequence-330586-        5834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent 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, Murphy-Royal C, Wen Y, Haidey JN, Molina RS, Aggarwal A, 
            Zhang S, Kamijo Y, Paquet ME, Podgorski K, Drobizhev M, Bains JS, 
            Lemieux MJ, Gordon GR, Campbell RE
  TITLE     A genetically encoded fluorescent biosensor for extracellular 
            L-lactate.
  JOURNAL   Nat Commun. 2021 Dec 6;12(1):7058. doi: 10.1038/s41467-021-27332-2.
  PUBMED    34873165
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: "2021-12-6"; volume: "12(1):7058. doi"; pages: " 
            10.1038/s41467-021-27332-2"
COMMENT     Sequence Label: pBAD-eLACCO1.1
FEATURES             Location/Qualifiers
     source          1..5834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..311
                     /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     234..253
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             357..374
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             378..410
                     /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(414..422,423..437)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     1244..1265
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1371..1392)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1632..1651)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(2261..2278)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2431..2517
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2609..2636
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2655..2746
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2747..2815,2816..3607)
                     /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(2965..2984)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3778..4366
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4267..4286
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4520..4537
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4552..4692
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4618..4637)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4778..4800
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4956..5834)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 acttttcata ctcccgccat tcagagaaga aaccaattgt ccatattgca tcagacattg
       61 ccgtcactgc gtcttttact ggctcttctc gctaaccaaa ccggtaaccc cgcttattaa
      121 aagcattctg taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca aaagtgtcta
      181 taatcacggc agaaaagtcc acattgatta tttgcacggc gtcacacttt gctatgccat
      241 agcattttta tccataagat tagcggatcc tacctgacgc tttttatcgc aactctctac
      301 tgtttctcca tacccgtttt ttgggctaac aggaggaatt aaccatgggg ggttctcatc
      361 atcatcatca tcatggtatg gctagcatga ctggtggaca gcaaatgggt cgggatctgt
      421 acgacgatga cgataaggat ccgagctcga gaatgttctc tcccttggca gtggcccagg
      481 ctcgacgcta taggtggaga attcaaaccg cgtgggatgc tggaacagta ggctacagtc
      541 tgttccaaaa gtttacggag cgggtaaagg aactgacgga tggacagctt gaggtccagc
      601 cttttccagc gggtgcagtc gtaggcacgt ttgatatgtt cgacgcggtc aagactggag
      661 ttttggacgg aatgaatcct ttcacgctgt tttgggcagg gaggatgccg gttacagctt
      721 tcctctccag ctacgccctc gggttggacc gacctgacca gtgggaaaca tggttttact
      781 ccctgggtgg cctcgacaac gcccgacgag cctttgccga gcaaggcctc ttctacgtgg
      841 gaccagttca gcacgacctc aatacaattc attcaaggaa gccgatccgg agatttgagg
      901 acttcaaagg tgtcaagctc cgagtgccgg gcggcatgat tgcggaagtt tttgcggccg
      961 caggagcttc caccgtcctc ttgcctgggg gggaggtcta ccccgctctg gaaagagggg
     1021 taatagattg gagccacaac gtctatatca tggccgacaa gcagaggaac ggcatcaagg
     1081 cgaacttcga gatccgccac aacatcgagg acggcggcgt gcagctcgcc taccactacc
     1141 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     1201 cccagaccaa actttcgaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1261 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggc ggtaccggag
     1321 ggagaatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     1381 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggtgag ggcgatgcca
     1441 cctccggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     1501 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     1561 tgaagcagca cgacttcttc aagtcggcca tgcccgaagg ctacatccag gagcgcacca
     1621 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     1681 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     1741 ggcataagct ggagtacagc tttaacgatg gtggtgcggc tgattttgtt gggcctgcag
     1801 taaactataa cttggggttc catcaggtgg caaagtacat cataatgggg cctccagaga
     1861 cgcctgccat tcatcaacct gtagacctca tggacttcac cataaacctt aatcggtggc
     1921 gcagccttcc aaagcctctg caagaaagat tcatcgcggc ggttcacgag tactcttgga
     1981 ttcactacgc aggaattcag aaggctaatt tggaggcgtg gccgaaatat cggcaggcag
     2041 gtgttgaggt catccggctg agcaacgagg atgttagaaa attcaggcga ctcgctattc
     2101 ccatttggtt taagtgggct aagatggata aatactcacg agaggcattc gcgtcccaac
     2161 tggagtatat gaaaggtata ggttacgtta cggacgagga attgaagggc ttgagcctgt
     2221 gaaagcttgg ctgttttggc ggatgagaga agattttcag cctgatacag attaaatcag
     2281 aacgcagaag cggtctgata aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac
     2341 ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc
     2401 cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac
     2461 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
     2521 ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg
     2581 ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttgcg
     2641 tttctacaaa ctcttttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     2701 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     2761 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     2821 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     2881 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     2941 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     3001 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     3061 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     3121 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     3181 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     3241 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     3301 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     3361 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     3421 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     3481 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     3541 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     3601 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     3661 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     3721 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3781 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaacaaccac cgctaccagc
     3841 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3901 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     3961 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4021 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4081 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4141 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4201 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4261 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4321 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4381 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     4441 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     4501 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     4561 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac
     4621 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
     4681 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     4741 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     4801 ttttcaccgt catcaccgaa acgcgcgagg cagcagatca attcgcgcgc gaaggcgaag
     4861 cggcatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac
     4921 tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca
     4981 ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta
     5041 aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata
     5101 ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag
     5161 cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag
     5221 caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg
     5281 tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct
     5341 tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc
     5401 ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc
     5461 gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca
     5521 tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga
     5581 tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa
     5641 acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata
     5701 taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc
     5761 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt
     5821 tgcgcttcag ccat
//
