LOCUS       sequence-410590-        6013 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 6013)
  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 6013)
  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-iLACCO1.1
FEATURES             Location/Qualifiers
     source          1..6013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     primer_bind     1875..1894
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2140..2728
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2193..2213)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2611..2622)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2760..3236
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2959..2978)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3276..3416
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3276..3405
                     /label=AAV2 ITR
     rep_origin      3491..3946
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3578..3597)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3788..3809
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3963..3982)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4082..4104
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4142..4160)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4228..4332
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4333..4401,4402..5193)
                     /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      5364..5952
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5853..5872
                     /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 cggggatcct
      841 ctagagtcga gaatggaaca aatcatagtc cagccgttga agaccctcat gggagacgac
      901 cctgattatt catttgatat tctggaggcg cggtacgcga tagaagcgtc aactgcctgg
      961 tacgctgcac tgcgagctac acccggggac aaagagaaaa ttcaactgtg ctacgaggct
     1021 acacggtccg aagatcccga tatagcttcc caagcagacg taaggttcca cctggcgatt
     1081 gctgaagcaa gccataacat agtactgctt cagacaatgc gaggcttttt tgatgttttg
     1141 cagtcttcag tcaagcacag tcgacagcgg atgtacttgg attgggtttc cgagcggatg
     1201 ttccccgagg acggcgccct gaagagcgag atcaagaagg ggctgaggct gaaggacggc
     1261 ggccactacg ccgccgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc
     1321 ggcgcctacg tcgtcgacat caagttggac atcgtgtccc acaacgagga ctacactatc
     1381 gtggaacagt gcgaacgcgc cgagggccgc cactccaccg gcggcgtgga cgagctgtac
     1441 aagggaggta caggcgggag tctggtgagc aagggcgagg aagatgacat ggccatcgtc
     1501 aaggagttca tgcgctacaa ggtgcacatg gagggctccg taaacggcca cgagttcgag
     1561 atcgagggcg agggcgaggg ccgcccctac gaggcctttc agaccgctaa gctgaaggtg
     1621 accaagggtg gccccctgcc cttcgcctgg gacatcctat cccctcagtt catgtacggc
     1681 tccaaggcct acattaagca cccagccgac atccccgact acttcaagct gtccttcccc
     1741 gagggcttca ggtgggagcg cgtgatgtac ttcgaggacg gcggcattat tcacgttaac
     1801 caggactcct ccctgcagga cggcgtattc atctacaagg tgaagctgcg cggcaccaac
     1861 ttcccccccg acggccccgt aatgcagaag aagaccatgg gctgggaggc gaggccgccg
     1921 gtctacggtc aactcacaga acagcatcag gctgttatag acgccatctt tgccggcgat
     1981 gcggacggtg ctagaaaagc tatgatggct catctgagct tcgtccatac gactatgaaa
     2041 cgcttcgatg gagatcaggc ccgacacgca cgcatttcca gactgcctgg agagcataat
     2101 gagcactctc gggaaaagaa agcatgaaag cttatcgata atcaacctct ggattacaaa
     2161 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     2221 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     2281 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     2341 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     2401 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     2461 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     2521 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt
     2581 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     2641 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     2701 cggatctccc tttgggccgc ctccccgcat cgataccgag cgctgctcga gagatctacg
     2761 ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag
     2821 tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct
     2881 tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa
     2941 cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc
     3001 tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt
     3061 tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg
     3121 gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt
     3181 ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga
     3241 ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc
     3301 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     3361 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     3421 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     3481 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3541 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
     3601 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     3661 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     3721 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3781 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
     3841 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
     3901 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     3961 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     4021 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     4081 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     4141 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     4201 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4261 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4321 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4381 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4441 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4501 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     4561 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     4621 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     4681 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     4741 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     4801 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     4861 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     4921 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     4981 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5041 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5101 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5161 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     5221 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5281 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5341 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5401 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5461 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5521 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5581 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5641 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5701 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5761 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5821 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5881 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5941 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6001 tttgctcaca tgt
//
