LOCUS       sequence-410570-        5938 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control biosensor for intracellular L-lactate.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5938)
  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 5938)
  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-diLACCO1
FEATURES             Location/Qualifiers
     source          1..5938
                     /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)"
     primer_bind     717..736
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        717..735
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      760..769
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1800..1819
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2065..2653
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2118..2138)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2536..2547)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2685..3161
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2884..2903)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3201..3341
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3201..3330
                     /label=AAV2 ITR
     rep_origin      3416..3871
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3503..3522)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3713..3734
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3888..3907)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4007..4029
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4067..4085)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4153..4257
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4258..4326,4327..5118)
                     /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(4476..4495)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5289..5877
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5778..5797
                     /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 caccggtgag ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat
      721 acgactcact atagggagac ccaagctggc tagcgtgccg ccaccatggc ctcgagaatg
      781 gtacaaaaca tagtccagcc gttgaagacc ctcatgggag acgaccctga ttattcattt
      841 gatattctgg aggcgcggta cgcgatagaa gcgtctactg cctggtacgc agcactgcga
      901 gctacacccg gggacaaaga gaaaattcaa cagtgcttcg aggctacacg gtccgaagat
      961 cccgatatag cttcccaagc aaacgtaagg ttccacctgg cgattgctga agcaagccat
     1021 aacatagtac tgctacagac gatgcgaggc ttttttgatg ttttgcagtc ttcagtcaag
     1081 cacagtcgac agcggatgta cttggattgg gcttccgagc ggatgttccc cgaggacggc
     1141 gccctgagga gcgtgatcaa gaaggggctg aggctgaagg acggcggcca ctacgccgcc
     1201 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacgtcgtc
     1261 gacatcaagt tggacatcgt gtcccacaac gaggactaca ctatcgtgga acagtgcgaa
     1321 cgcgccgagg gccgccactc caccggcggc gtggacgagc tgtacaaggg aggtacaggc
     1381 gggagtctgg tgagcaaggg cgaggaggat gacatggcca tcgtcaagga gttcatgcgc
     1441 tacaaggtgc acatggaggg ctccgtaaac ggccacgagt tcgagatcga gggcgagggc
     1501 gagggccgcc cctacgaggc ctttcagacc gctaagctga aggtgaccaa gggtggcccc
     1561 ctgcccttcg cctgggacat cctatcccct cagttcatgt acggctccaa ggcctacatt
     1621 aagcacccag ccgatatccc cgactacttc aagctgtcct tccccgaggg cttcaggtgg
     1681 gagcgcgtga tgtacttcga ggacggcggc attattcacg ttaaccagga ctcctccctg
     1741 caggacggcg tatttatcta caaggtgaag ctgcgcggca ccaacttccc ccccgacggc
     1801 cccgtaatgc agaagaagac catgggctgg gaggcgaggc cgccggtcta cggtcaactc
     1861 acagaacagc atcaggcagt tatagacgcc atctttgccg gcgatgcgga cggtgctaga
     1921 aaagctatga tggctcatct gagcttcgtc cattcgacta tgcaacgctt cgatggagat
     1981 caggcccgac acgcacgcat ttccagactg cctggagagc ataatgagca ctctcgggaa
     2041 aagaaagcat gaaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     2101 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     2161 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     2221 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     2281 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     2341 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     2401 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     2461 tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc
     2521 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     2581 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     2641 gccgcctccc cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg catccctgtg
     2701 acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg
     2761 tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg
     2821 ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg
     2881 gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg
     2941 cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat
     3001 gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg
     3061 ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc
     3121 tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca
     3181 cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     3241 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     3301 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     3361 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     3421 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3481 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3541 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3601 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     3661 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3721 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
     3781 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3841 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     3901 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     3961 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     4021 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     4081 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     4141 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4201 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4261 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4321 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4381 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4441 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4501 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4561 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4621 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     4681 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4741 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     4801 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     4861 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     4921 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     4981 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5041 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5101 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5161 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5221 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5281 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5341 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5401 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5461 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5521 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5581 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5641 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5701 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5761 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5821 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5881 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
