LOCUS       sequence-330589-        5834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent biosensor control 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-deLACCO
FEATURES             Location/Qualifiers
     source          1..5834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             46..63
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             67..99
                     /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(103..111,112..126)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     933..954
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1060..1081)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1321..1340)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(1950..1967)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2120..2206
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2298..2325
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2344..2435
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2436..2504,2505..3296)
                     /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(2654..2673)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3467..4055
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3956..3975
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4209..4226
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4241..4381
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4307..4326)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4467..4489
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4645..5523)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        5550..5834
                     /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     5757..5776
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 acccgttttt tgggctaaca ggaggaatta accatggggg gttctcatca tcatcatcat
       61 catggtatgg ctagcatgac tggtggacag caaatgggtc gggatctgta cgacgatgac
      121 gataaggatc cgagctcgag aatgttctct cccttggcag tggcccaggc tcgacgctat
      181 aggtggagga ttcaaaccgc gtgggatgct ggaacagtag gctacagtct gttccaaaag
      241 tttacggagc gggtaaagga actgacggat ggacagcttg aggtccagcc ttttccagcg
      301 ggtgcagtcg taggcacgtt tgatatgttc gacgcggtca agactggagt tttggacgga
      361 atgaatcctt tcacgctgtt ttgggcaggg aggatgccgg ttacagcttt cctctccagc
      421 tacgccctcg ggttggaccg acctgaccag tgggaaacat ggttttactc cctgggtggc
      481 ctcgacaacg cccgacgagc ctttgccgag caaggcctct tctacgtggg accagttcag
      541 cacgacctca atacaattca ttcaaggaag ccgatccgga gatttgagga cttcaaaggt
      601 gtcaagctcc gagtgccggg cggcatgatt gcggaagttt ttgcggccgc aggagcttcc
      661 accgtcctct tgcctggggg ggaggtctac cccgctctgg aaagaggggt aatagattgg
      721 agccacaacg tctatatcat ggccgacaag cagaggaacg gcatcaaggc gaacttcgag
      781 atccgccaca acatcgagga cggcggcgtg cagctcgcct accactacca gcagaacacc
      841 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagaccaaa
      901 ctttcgaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
      961 gccgggatca ctctcggcat ggacgagctg tacaagggcg gtaccggagg gagaatggtg
     1021 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     1081 gtaaacggcc acaagttcag cgtgtccggc gagggtgagg gcgatgccac ctccggcaag
     1141 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     1201 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     1261 gacttcttca agtcggccat gcccgaaggc tacatccagg agcgcaccat cttcttcaag
     1321 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1381 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcataagctg
     1441 gagtacagct ttaacgatgg tggtgcggct aattttgttg ggcctgcagt aaactataac
     1501 ttggggttcc atcaggtggc aaagtacatc ataatggggc ctccagagac gcctgccatt
     1561 catcaacctg tagacctcat ggacttcacc ataaacctta atcggtggcg cagccttcca
     1621 aagcctctgc aagaaagatt catcgcggcg gttcacgagt actcttggat tcactacgca
     1681 ggcattcaga aggctaattt ggaggcgtgg ccgaaatatc ggcaggcagg tgttgaggtc
     1741 atccggctga gcaacgagga tgttagaaaa ttcaggcgac tcgctattcc catttggttt
     1801 aagtgggcta agatggataa atactcacga gaggcattcg cgtcccaact ggagtatatg
     1861 aaaggtatag gttacgttac ggacgaggaa ttgaagggct tgagcctgtg aaagcttggc
     1921 tgttttggcg gatgagagaa gattttcagc ctgatacaga ttaaatcaga acgcagaagc
     1981 ggtctgataa aacagaattt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg
     2041 ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga
     2101 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     2161 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga
     2221 tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc
     2281 caggcatcaa attaagcaga aggccatcct gacggatggc ctttttgcgt ttctacaaac
     2341 tcttttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2401 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     2461 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     2521 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     2581 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     2641 cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac
     2701 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     2761 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     2821 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     2881 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     2941 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     3001 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     3061 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     3121 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     3181 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     3241 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     3301 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     3361 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     3421 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3481 ttctgcgcgt aatctgctgc ttgcaaacaa aacaaccacc gctaccagcg gtggtttgtt
     3541 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3601 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3661 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3721 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3781 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3841 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3901 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3961 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4021 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4081 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     4141 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     4201 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     4261 ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg
     4321 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
     4381 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     4441 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     4501 atcaccgaaa cgcgcgaggc agcagatcaa ttcgcgcgcg aaggcgaagc ggcatgcata
     4561 atgtgcctgt caaatggacg aagcagggat tctgcaaacc ctatgctact ccgtcaagcc
     4621 gtcaattgtc tgattcgtta ccaattatga caacttgacg gctacatcat tcactttttc
     4681 ttcacaaccg gcacggaact cgctcgggct ggccccggtg cattttttaa atacccgcga
     4741 gaaatagagt tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag gcatccgggt
     4801 ggtgctcaaa agcagcttcg cctggctgat acgttggtcc tcgcgccagc ttaagacgct
     4861 aatccctaac tgctggcgga aaagatgtga cagacgcgac ggcgacaagc aaacatgctg
     4921 tgcgacgctg gcgatatcaa aattgctgtc tgccaggtga tcgctgatgt actgacaagc
     4981 ctcgcgtacc cgattatcca tcggtggatg gagcgactcg ttaatcgctt ccatgcgccg
     5041 cagtaacaat tgctcaagca gatttatcgc cagcagctcc gaatagcgcc cttccccttg
     5101 cccggcgtta atgatttgcc caaacaggtc gctgaaatgc ggctggtgcg cttcatccgg
     5161 gcgaaagaac cccgtattgg caaatattga cggccagtta agccattcat gccagtaggc
     5221 gcgcggacga aagtaaaccc actggtgata ccattcgcga gcctccggat gacgaccgta
     5281 gtgatgaatc tctcctggcg ggaacagcaa aatatcaccc ggtcggcaaa caaattctcg
     5341 tccctgattt ttcaccaccc cctgaccgcg aatggtgaga ttgagaatat aacctttcat
     5401 tcccagcggt cggtcgataa aaaaatcgag ataaccgttg gcctcaatcg gcgttaaacc
     5461 cgccaccaga tgggcattaa acgagtatcc cggcagcagg ggatcatttt gcgcttcagc
     5521 catacttttc atactcccgc cattcagaga agaaaccaat tgtccatatt gcatcagaca
     5581 ttgccgtcac tgcgtctttt actggctctt ctcgctaacc aaaccggtaa ccccgcttat
     5641 taaaagcatt ctgtaacaaa gcgggaccaa agccatgaca aaaacgcgta acaaaagtgt
     5701 ctataatcac ggcagaaaag tccacattga ttatttgcac ggcgtcacac tttgctatgc
     5761 catagcattt ttatccataa gattagcgga tcctacctga cgctttttat cgcaactctc
     5821 tactgtttct ccat
//
