LOCUS       sequence-410586-        6051 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 6051)
  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 6051)
  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-gfaABC1D-R-iLACCO1.2
FEATURES             Location/Qualifiers
     source          1..6051
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      841..850
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1869..1888
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2178..2766
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2231..2251)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2649..2660)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2798..3274
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2997..3016)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3314..3454
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3314..3443
                     /label=AAV2 ITR
     rep_origin      3529..3984
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3616..3635)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3826..3847
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4001..4020)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4120..4142
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4180..4198)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4266..4370
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4371..4439,4440..5231)
                     /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"
     rep_origin      5402..5990
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5891..5910
                     /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 gcgtggatct aacatatcct ggtgtggagt aggggacgct
      181 gctctgacag aggctcgggg gcctgagctg gctctgtgag ctggggagga ggcagacagc
      241 caggccttgt ctgcaagcag acctggcagc attgggctgg ccgcccccca gggcctcctc
      301 ttcatgccca gtgaatgact caccttggca cagacacaat gttcggggtg ggcacagtgc
      361 ctgcttcccg ccgcacccca gcccccctca aatgccttcc gagaagccca ttgagcaggg
      421 ggcttgcatt gcaccccagc ctgacagcct ggcatcttgg gataaaagca gcacagcccc
      481 ctaggggctg cccttgctgt gtggcgccac cggcggtgga gaacaaggct ctattcagcc
      541 tgtgcccagg aaaggggatc aggggatgcc caggcatgga cagtgggtgg caggggggga
      601 gaggagggct gtctgcttcc cagaagtcca aggacacaaa tgggtgaggg gagagctctc
      661 cccatagctg ggctgcggcc caaccccacc ccctcaggct atgccagggg gtgttgccag
      721 gggcacccgg gcatcgccag tctagcccac tccttcataa agccctcgca tcccaggagc
      781 gagcagagcc agagcaggtt ggagaggaga cgcatcacct ccgctgctcg cggggatccc
      841 gccaccatgg aacaaaacat agtccagccg ttgaagaccc tcatgggaga cgaccctgat
      901 tattcatttg atattctgga ggcgcggtac gcgatagaag cgtcaactgc ctggtacgct
      961 gcactgcgag ctacacccgg ggacaaagag aaaattcaac tgtgcttcga ggctacacgg
     1021 tccgaagatc ccgatatagc ttcccaagca gacgtaaggt tccacctggc gattgctgaa
     1081 gcaagccata acatagttct gcttcagaca atgcgaggct tttttgatgt tttgcagtct
     1141 tcagtcaagc acagtcggca gcggatgtac ttggattggg tttccgagcg gatgttcccc
     1201 gaggacggcg ccctgaagag cgagatcaag aaggggctga ggctgaagga cggcggccac
     1261 tacgccgccg aggtcaagac cacttacaag gccaagaagc ccgtgcagct gcccggcgcc
     1321 tacgtcgtcg acatcaagtt ggacatcgtg tcccacaacg aggactacac tatcgtggaa
     1381 cagtgcgaac gcgccgaggg ccgccactcc accggcggcg tggacgagct gtacaaggga
     1441 ggtacaggcg ggagtctggt gagcaagggc gaggaggatg acatggccat cgtcaaggag
     1501 ttcatgcgct tcaaggtgca catggagggc tccgtaaacg gccacgagtt cgagatcgag
     1561 ggcgagggcg agggccgccc ctacgaggcc tttcagaccg ctaagctgaa ggtgaccaag
     1621 ggtggccccc tgcccttcgc ctgggacatc ctatcccctc agttcatgta cggctccaag
     1681 gcctacatta agcacccagc cgacatcccc gactacttca agctgtcctt ccccgagggc
     1741 ttcaggtggg agcgcgtgat gtacttcgag gacggcggca ttattcacgt taaccaggac
     1801 tcctccctgc aggacggcgt attcatctac aaggtgaagc tgcgcggcac caacttcccc
     1861 cccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcgaggcc gccggtctac
     1921 ggtcaactca cagaacagca tcaggcagtt atagacgcca tctttgccgg cgatgcggac
     1981 ggtgctagaa aagctatgat ggctcatctg agcttcatcc atacgactat gaaacgcttc
     2041 gatggagatc aggcccgaca cgcacgcatt tccagactgc ctggagagca taatgagcac
     2101 tctcgggaaa agaaagcatg agcggccgct cgacctcgag gggcccggta cccgaattcg
     2161 atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     2221 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     2281 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     2341 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     2401 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     2461 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     2521 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     2581 cctttccttg gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct
     2641 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     2701 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     2761 ccccgcatcg ataccgagcg ctgctcgaga gatctacggg tggcatccct gtgacccctc
     2821 cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat
     2881 aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag
     2941 gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta
     3001 ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg
     3061 ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac
     3121 caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct
     3181 ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt
     3241 acaggcgtga accactgctc ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg
     3301 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     3361 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     3421 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     3481 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     3541 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3601 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3661 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3721 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     3781 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3841 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     3901 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     3961 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     4021 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4081 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     4141 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     4201 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     4261 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4321 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4381 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4441 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4501 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4561 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     4621 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4681 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4741 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     4801 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     4861 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     4921 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     4981 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5041 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5101 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5161 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5221 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5281 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5341 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5401 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5461 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5521 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     5581 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5641 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5701 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5761 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5821 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5881 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5941 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6001 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
