LOCUS       sequence-439061-        5872 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 5872)
  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 5872)
  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-iLACCO1.1
FEATURES             Location/Qualifiers
     source          1..5872
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..455
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     primer_bind     1567..1586
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    1832..2420
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1885..1905)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2303..2314)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2452..2928
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2651..2670)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2968..3108
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2968..3097
                     /label=AAV2 ITR
     rep_origin      3183..3638
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3270..3289)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3480..3501
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3655..3674)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3774..3796
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3834..3852)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3920..4024
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4025..4093,4094..4885)
                     /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(4243..4262)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5056..5644
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5545..5564
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5706..5835
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gcgtatgagt gcaagtgggt tttaggacca ggatgaggcg gggtgggggt gcctacctga
       61 cgaccgaccc cgacccactg gacaagcacc caacccccat tccccaaatt gcgcatcccc
      121 tatcagagag ggggagggga aacaggatgc ggcgaggcgc gtgcgcactg ccagcttcag
      181 caccgcggac agtgccttcg cccccgcctg gcggcgcgcg ccaccgccgc ctcagcactg
      241 aaggcgcgct gacgtcactc gccggtcccc cgcaaactcc ccttcccggc caccttggtc
      301 gcgtccgcgc cgccgccggc ccagccggac cgcaccacgc gaggcgcgag ataggggggc
      361 acgggcgcga ccatctgcgc tgcggcgccg gcgactcagc gctgcctcag tctgcggtgg
      421 gcagcggagg agtcgtgtcg tgcctgagag cgcagtcgag aaggtaccgg atcggacgaa
      481 taacgcgtgg atccaccggt gagctcccgc ggcggccgct ctagattagt ctgcagattc
      541 gagaatggaa caaatcatag tccagccgtt gaagaccctc atgggagacg accctgatta
      601 ttcatttgat attctggagg cgcggtacgc gatagaagcg tcaactgcct ggtacgctgc
      661 actgcgagct acacccgggg acaaagagaa aattcaactg tgctacgagg ctacacggtc
      721 cgaagatccc gatatagctt cccaagcaga cgtaaggttc cacctggcga ttgctgaagc
      781 aagccataac atagtactgc ttcagacaat gcgaggcttt tttgatgttt tgcagtcttc
      841 agtcaagcac agtcgacagc ggatgtactt ggattgggtt tccgagcgga tgttccccga
      901 ggacggcgcc ctgaagagcg agatcaagaa ggggctgagg ctgaaggacg gcggccacta
      961 cgccgccgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
     1021 cgtcgtcgac atcaagttgg acatcgtgtc ccacaacgag gactacacta tcgtggaaca
     1081 gtgcgaacgc gccgagggcc gccactccac cggcggcgtg gacgagctgt acaagggagg
     1141 tacaggcggg agtctggtga gcaagggcga ggaagatgac atggccatcg tcaaggagtt
     1201 catgcgctac aaggtgcaca tggagggctc cgtaaacggc cacgagttcg agatcgaggg
     1261 cgagggcgag ggccgcccct acgaggcctt tcagaccgct aagctgaagg tgaccaaggg
     1321 tggccccctg cccttcgcct gggacatcct atcccctcag ttcatgtacg gctccaaggc
     1381 ctacattaag cacccagccg acatccccga ctacttcaag ctgtccttcc ccgagggctt
     1441 caggtgggag cgcgtgatgt acttcgagga cggcggcatt attcacgtta accaggactc
     1501 ctccctgcag gacggcgtat tcatctacaa ggtgaagctg cgcggcacca acttcccccc
     1561 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcgaggccgc cggtctacgg
     1621 tcaactcaca gaacagcatc aggctgttat agacgccatc tttgccggcg atgcggacgg
     1681 tgctagaaaa gctatgatgg ctcatctgag cttcgtccat acgactatga aacgcttcga
     1741 tggagatcag gcccgacacg cacgcatttc cagactgcct ggagagcata atgagcactc
     1801 tcgggaaaag aaagcatgaa agcttatcga taatcaacct ctggattaca aaatttgtga
     1861 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     1921 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     1981 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     2041 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     2101 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     2161 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     2221 ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg
     2281 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     2341 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     2401 cctttgggcc gcctccccgc atcgataccg agcgctgctc gagagatcta cgggtggcat
     2461 ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc
     2521 agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat
     2581 attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg
     2641 gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca
     2701 atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc
     2761 caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc
     2821 atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc
     2881 aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttgtag
     2941 gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     3001 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     3061 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     3121 ggtattttct ccctacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     3181 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3241 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3301 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     3361 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     3421 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3481 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     3541 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     3601 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     3661 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     3721 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     3781 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     3841 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     3901 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3961 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4021 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4081 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4141 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4201 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4261 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     4321 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     4381 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     4441 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     4501 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     4561 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     4621 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     4681 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     4741 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     4801 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     4861 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4921 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4981 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5041 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5101 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5161 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5221 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5281 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5341 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5401 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5461 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5521 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5581 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5641 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5701 catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga
     5761 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
     5821 tcactagggg ttcctgcggc cgcacgcgtg tgtctagact gcagagggcc ct
//
