LOCUS       sequence-414442-        7388 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Biosensor for extracellular L-lactate.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7388)
  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 7388)
  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-CaMKIIa-HA-eLACCO2.1-NGR
FEATURES             Location/Qualifiers
     source          1..7388
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   71..200
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        219..1507
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1542..1551
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2532..2553)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2793..2812)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    3585..4173
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3638..3658)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4056..4067)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4205..4681
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4404..4423)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4721..4861
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4721..4850
                     /label=AAV2 ITR
     rep_origin      4936..5391
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5023..5042)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5233..5254
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5408..5427)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5527..5549
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5587..5605)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5673..5777
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5778..5846,5847..6638)
                     /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(5996..6015)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6809..7388,1..9)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7298..7317
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
       61 gctcacatgt cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg
      121 ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa
      181 ctccatcact aggggttcct gcggccgcac gcgtttaaca ttatggcctt aggtcacttc
      241 atctccatgg ggttcttctt ctgattttct agaaaatgag atgggggtgc agagagcttc
      301 ctcagtgacc tgcccagggt cacatcagaa atgtcagagc tagaacttga actcagatta
      361 ctaatcttaa attccatgcc ttgggggcat gcaagtacga tatacagaag gagtgaactc
      421 attagggcag atgaccaatg agtttaggaa agaagagtcc agggcagggt acatctacac
      481 cacccgccca gccctgggtg agtccagcca cgttcacctc attatagttg cctctctcca
      541 gtcctacctt gacgggaagc acaagcagaa actgggacag gagccccagg agaccaaatc
      601 ttcatggtcc ctctgggagg atgggtgggg agagctgtgg cagaggcctc aggaggggcc
      661 ctgctgctca gtggtgacag ataggggtga gaaagcagac agagtcattc cgtcagcatt
      721 ctgggtctgt ttggtacttc ttctcacgct aaggtggcgg tgtgatatgc acaatggcta
      781 aaaagcaggg agagctggaa agaaacaagg acagagacag aggccaagtc aaccagacca
      841 attcccagag gaagcaaaga aaccattaca gagactacaa gggggaaggg aaggagagat
      901 gaattagctt cccctgtaaa ccttagaacc cagctgttgc cagggcaacg gggcaatacc
      961 tgtctcttca gaggagatga agttgccagg gtaactacat cctgtctttc tcaaggacca
     1021 tcccagaatg tggcacccac tagccgttac catagcaact gcctctttgc cccacttaat
     1081 cccatcccgt ctgttaaaag ggccctatag ttggaggtgg gggaggtagg aagagcgatg
     1141 atcacttgtg gactaagttt gttcgcatcc ccttctccaa ccccctcagt acatcaccct
     1201 gggggaacag ggtccacttg ctcctgggcc cacacagtcc tgcagtattg tgtatataag
     1261 gccagggcaa agaggagcag gttttaaagt gaaaggcagg caggtgttgg ggaggcagtt
     1321 accggggcaa cgggaacagg gcgtttcgga ggtggttgcc atggggacct ggatgctgac
     1381 gaaggctcgc gaggctgtga gcagccacag tgccctgctc agaagcccca agctcgtcag
     1441 tcaagccggt tctccgtttg cactcaggag cacgggcagg cgagtggccc ctagttctgg
     1501 gggcagctct agagcggtac cggatccctg gctagcgtgc cgccaccatg gcctcgagaa
     1561 tgaagaccat catcgccctg tcctacatct tctgcctggt gctgggcaga tctatgttct
     1621 ctcccttggc agtggcccag gctcgacgct ataggtggcg cattcaaacc gcgtgggatg
     1681 ccggaacagt aggctacagt ctgttccaaa agtttacgga gcgggtaaag gaactgacgg
     1741 atggacagct tgaggtccag ccttttccag cgagtgcagt cgtaggcacg tttgatatgt
     1801 tcgacgcggt caagactgga gttttggacg gaatgaatcc tttcacgatc tattgggcag
     1861 ggaggatgcc ggttacagct ttcctctcca gctacgccct cgggttggac cgacctgacc
     1921 agtgggaaac atggttttac tccctgggtg gcctcgacaa cgcccgacga gcctttgccg
     1981 agcaaggcct cttctacgtg ggaccagttc agcacgacct caatacaatt cattcaaaga
     2041 agccgatccg gagatttgag gacttcaaag gtgtcaagct ccgagtgccg ggcggcatga
     2101 ttgcggaagt ttttgcggcc gcaggagctt ccaccgtcct cttgcctggg ggggaggtca
     2161 accccgctct ggaaagaggg gtaatagatt ggagccacaa cgtccatatc atggccgaca
     2221 agcagaggaa cggcatcaag gcgaacttcg agatccgcca cagcaccgag gacggcggcg
     2281 tgcagctcgc ctaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     2341 ccgacaacca ctacctgagc acccagacca aacttacgaa agaccccaac gagaagcgcg
     2401 atcacatggt cctgctggag tacgtgaccg ccgccgggat cactcgcggc atggacgagc
     2461 tgtacaaggg cggtaccgga gggagaatgg tgagcaaggg cgaggagctg ttcaccgggg
     2521 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     2581 gcgagggtga gggcgatgcc acctccggca agctgtccct gaagttcatc agcaccaccg
     2641 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     2701 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtcggcc atgcccgaag
     2761 gctacatcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     2821 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     2881 aggaggacgg caacatcctg gggcataagc tggagtacag ctttaacgat ggtggtgcgg
     2941 ctgattttgt tgggcctgca gtaaactata acttggggtt ccatcaggtg gcaaagtaca
     3001 tcataatggg gcctccagag acgcctgcca ttcatcaacc tgtagacctc atggacttca
     3061 ccataaacct taatcggtgg cgcagccttc caaagcccct gcaagaaaga ttcatcgcgg
     3121 cggttcacga gtactcttgg attcactacg caggcattca gaaggctaat ttggaggcgt
     3181 ggccgaaata tcggcaggca ggtgttgagg tcatccggct gagcaacgag gatgttagaa
     3241 aattcaggcg actcgctatt cccatttggt ttaagtgggc taagatggat aactactcac
     3301 gagaggcatt cgcgtctcaa ctggagtata tgaaaggtat aggttacgtt acggacgagg
     3361 aattgaaggg cttgggcctg ggctccacct ccggctccgg caagcccggc tccggcgagg
     3421 gctccaccaa gggcgaattc gcaggtagtg gtgcatctgg aacaggagat gcagaaggat
     3481 caggtgccct tcctgccctt gcatgtagtc tggctccact gggactggca ttggttctgt
     3541 ggacggtgtt gggcccctgt tagactagtg caaagcttat cgataatcaa cctctggatt
     3601 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     3661 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     3721 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     3781 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     3841 ccacctgtca gctcctttcc ggaactttcg ctttccccct ccctattgcc acggcggaac
     3901 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     3961 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct
     4021 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     4081 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     4141 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgagcgctg ctcgagagat
     4201 ctacgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac
     4261 tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt
     4321 gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa
     4381 gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc
     4441 ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga
     4501 gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag
     4561 acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc
     4621 accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct
     4681 tctgattttg taggtaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag
     4741 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     4801 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     4861 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     4921 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     4981 cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc
     5041 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     5101 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     5161 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     5221 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct
     5281 attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga
     5341 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     5401 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     5461 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     5521 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     5581 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     5641 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     5701 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     5761 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5821 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5881 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5941 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     6001 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     6061 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     6121 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     6181 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     6241 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     6301 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     6361 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     6421 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     6481 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     6541 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     6601 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     6661 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6721 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6781 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6841 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6901 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6961 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     7021 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     7081 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     7141 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     7201 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     7261 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     7321 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     7381 ggcggagc
//
