LOCUS       sequence-410584-        5844 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Biosensor for intracellular L-lactate.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5844)
  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 5844)
  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: pAAV-hSyn-R-iLACCO1.2
FEATURES             Location/Qualifiers
     source          1..5844
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      663..672
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1691..1710
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    1971..2559
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2024..2044)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2442..2453)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2591..3067
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2790..2809)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3107..3247
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3107..3236
                     /label=AAV2 ITR
     rep_origin      3322..3777
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3409..3428)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3619..3640
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3794..3813)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3913..3935
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3973..3991)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4059..4163
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4164..4232,4233..5024)
                     /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     complement(4382..4401)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5195..5783
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5684..5703
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc ggacgaataa cgcgtggatc
      661 ccgccaccat ggaacaaaac atagtccagc cgttgaagac cctcatggga gacgaccctg
      721 attattcatt tgatattctg gaggcgcggt acgcgataga agcgtcaact gcctggtacg
      781 ctgcactgcg agctacaccc ggggacaaag agaaaattca actgtgcttc gaggctacac
      841 ggtccgaaga tcccgatata gcttcccaag cagacgtaag gttccacctg gcgattgctg
      901 aagcaagcca taacatagtt ctgcttcaga caatgcgagg cttttttgat gttttgcagt
      961 cttcagtcaa gcacagtcgg cagcggatgt acttggattg ggtttccgag cggatgttcc
     1021 ccgaggacgg cgccctgaag agcgagatca agaaggggct gaggctgaag gacggcggcc
     1081 actacgccgc cgaggtcaag accacttaca aggccaagaa gcccgtgcag ctgcccggcg
     1141 cctacgtcgt cgacatcaag ttggacatcg tgtcccacaa cgaggactac actatcgtgg
     1201 aacagtgcga acgcgccgag ggccgccact ccaccggcgg cgtggacgag ctgtacaagg
     1261 gaggtacagg cgggagtctg gtgagcaagg gcgaggagga tgacatggcc atcgtcaagg
     1321 agttcatgcg cttcaaggtg cacatggagg gctccgtaaa cggccacgag ttcgagatcg
     1381 agggcgaggg cgagggccgc ccctacgagg cctttcagac cgctaagctg aaggtgacca
     1441 agggtggccc cctgcccttc gcctgggaca tcctatcccc tcagttcatg tacggctcca
     1501 aggcctacat taagcaccca gccgacatcc ccgactactt caagctgtcc ttccccgagg
     1561 gcttcaggtg ggagcgcgtg atgtacttcg aggacggcgg cattattcac gttaaccagg
     1621 actcctccct gcaggacggc gtattcatct acaaggtgaa gctgcgcggc accaacttcc
     1681 cccccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcgagg ccgccggtct
     1741 acggtcaact cacagaacag catcaggcag ttatagacgc catctttgcc ggcgatgcgg
     1801 acggtgctag aaaagctatg atggctcatc tgagcttcat ccatacgact atgaaacgct
     1861 tcgatggaga tcaggcccga cacgcacgca tttccagact gcctggagag cataatgagc
     1921 actctcggga aaagaaagca tgagcggccg ctcgaatcaa gcttatcgat aatcaacctc
     1981 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     2041 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     2101 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     2161 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     2221 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     2281 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     2341 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg
     2401 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     2461 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     2521 ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccga gcgctgctcg
     2581 agagatctac gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt
     2641 tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga
     2701 ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt
     2761 tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc
     2821 acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc
     2881 tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg
     2941 gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat
     3001 ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc
     3061 tgtccttctg attttgtagg taaccacgtg cggaccgagc ggccgcagga acccctagtg
     3121 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     3181 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     3241 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     3301 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3361 tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt
     3421 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3481 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     3541 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3601 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct
     3661 cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg
     3721 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     3781 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     3841 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     3901 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     3961 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     4021 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4081 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4141 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     4201 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4261 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     4321 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     4381 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     4441 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4501 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     4561 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     4621 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4681 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     4741 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     4801 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4861 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     4921 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4981 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     5041 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5101 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5161 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5221 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5281 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5341 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5401 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5461 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5521 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5581 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5641 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5701 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5761 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5821 tttgctggcc ttttgctcac atgt
//
