LOCUS       sequence-331500-        6456 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent sensor for extracellular lactate.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6456)
  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 6456)
  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: pAAV-CAG-eLACCO1.1
FEATURES             Location/Qualifiers
     source          1..6456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(28..456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(138..159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     350..369
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   528..657
                     /label=AAV2 ITR
     promoter        968..1245
                     /label=chicken beta-actin promoter
     primer_bind     2281..2302
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2408..2429)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2669..2688)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    3449..4037
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3502..3522)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3920..3931)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4080..4201
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4091..4110)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4145..4164
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(4707..5295)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4787..4806)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5466..6257,6258..6326))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6089..6108
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6327..6431)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
       61 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
      121 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
      181 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
      241 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
      301 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
      361 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
      421 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
      481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg cgcgctcgct
      541 cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt cgcccggcct
      601 cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg ggttccttgt
      661 agttaatgat taacccgcca tgctacttat ctacgtagcc atgctctagg aagagtacca
      721 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      781 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      841 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      901 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      961 accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca
     1021 cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg
     1081 gggggggggc gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg
     1141 agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg
     1201 cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc
     1261 tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg
     1321 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag
     1381 cgctaccgga ctcagatctg atatcaccat gggaatccaa ggagggtctg tcctgttcgg
     1441 gctgctgctc gtcctggctg tcttctgcca ttcaggtcat agcagatcta tgttctctcc
     1501 cttggcagtg gcccaggctc gacgctatag gtggaggatt caaaccgcgt gggatgctgg
     1561 aacagtaggc tacagtctgt tccaaaagtt tacggagcgg gtaaaggaac tgacggatgg
     1621 acagcttgag gtccagcctt ttccagcggg tgcagtcgta ggcacgtttg atatgttcga
     1681 cgcggtcaag actggagttt tggacggaat gaatcctttc acgctgtttt gggcagggag
     1741 gatgccggtt acagctttcc tctccagcta cgccctcggg ttggaccgac ctgaccagtg
     1801 ggaaacatgg ttttactccc tgggtggcct cgacaacgcc cgacgagcct ttgccgagca
     1861 aggcctcttc tacgtgggac cagttcagca cgacctcaat acaattcatt caaggaagcc
     1921 gatccggaga tttgaggact tcaaaggtgt caagctccga gtgccgggcg gcatgattgc
     1981 ggaagttttt gcggccgcag gagcttccac cgtcctcttg cctggggggg aggtctaccc
     2041 cgctctggaa agaggggtaa tagattggag ccacaacgtc tatatcatgg ccgacaagca
     2101 gaggaacggc atcaaggcga acttcgagat ccgccacaac atcgaggacg gcggcgtgca
     2161 gctcgcctac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     2221 caaccactac ctgagcaccc agaccaaact ttcgaaagac cccaacgaga agcgcgatca
     2281 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     2341 caagggcggt accggaggga gaatggtgag caagggcgag gagctgttca ccggggtggt
     2401 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     2461 gggtgagggc gatgccacct ccggcaagct gaccctgaag ttcatctgca ccaccggcaa
     2521 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     2581 ccgctacccc gaccacatga agcagcacga cttcttcaag tcggccatgc ccgaaggcta
     2641 catccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     2701 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     2761 ggacggcaac atcctggggc ataagctgga gtacagcttt aacgatggtg gtgcggctga
     2821 ttttgttggg cctgcagtaa actataactt ggggttccat caggtggcaa agtacatcat
     2881 aatggggcct ccagagacgc ctgccattca tcaacctgta gacctcatgg acttcaccat
     2941 aaaccttaat cggtggcgca gccttccaaa gcctctgcaa gaaagattca tcgcggcggt
     3001 tcacgagtac tcttggattc actacgcagg cattcagaag gctaatttgg aggcgtggcc
     3061 gaaatatcgg caggcaggtg ttgaggtcat ccggctgagc aacgaggatg ttagaaaatt
     3121 caggcgactc gctattccca tttggtttaa gtgggctaag atggataaat actcacgaga
     3181 ggcattcgcg tcccaactgg agtatatgaa aggtataggt tacgttacgg acgaggaatt
     3241 gaagggcttg agcctgggct ccacctccgg ctccggcaag cccggctccg gcgagggctc
     3301 caccaagggc gaattccttg aaaatggtgg gacatcctta tcagagaaaa cagttcttct
     3361 gctggtgact ccatttctgg cagcagcctg gagccttcat ccctaaggat cctctagagt
     3421 cgacctgcag gcatgcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag
     3481 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     3541 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     3601 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     3661 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     3721 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     3781 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     3841 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     3901 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     3961 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     4021 ttgggccgcc tccccgcatc gataccgtcg acccgggcgg ccgcttcgag cagacatgat
     4081 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     4141 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     4201 taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt
     4261 ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatcttc ctagagcatg
     4321 gctacgtaga taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg
     4381 agttggccac tccctctctg cgcgctcgct cgctcactga ggacgcccgg gctttgcccg
     4441 ggcggcctca gtgagcgagc gagcgcgcag ctgcattaat gaatcggcca acgcgcgggg
     4501 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4561 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4621 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4681 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4741 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4801 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4861 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4921 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4981 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5041 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5101 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5161 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5221 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5281 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5341 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5401 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5461 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5521 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5581 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5641 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5701 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5761 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5821 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5881 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5941 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6001 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6061 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6121 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6181 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6241 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6301 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6361 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6421 ggggttccgc gcacatttcc ccgaaaagtg ccacct
//
