LOCUS       sequence-331493-        6602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Neuronal expression of fluorescent sensor for extracellular lactate.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6602)
  AUTHORS   Nasu Y, Murphy-Royal C, Wen Y, Haidey JN, Molina RS, Aggarwal A, 
            Zhang S, Kamijo Y, Paquet ME, Podgorski K, Drobizhev M, Bains JS, 
            Lemieux MJ, Gordon GR, Campbell RE
  TITLE     A genetically encoded fluorescent biosensor for extracellular 
            L-lactate.
  JOURNAL   Nat Commun. 2021 Dec 6;12(1):7058. doi: 10.1038/s41467-021-27332-2.
  PUBMED    34873165
REFERENCE   2  (bases 1 to 6602)
  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: "2021-12-6"; volume: "12(1):7058. doi"; pages: " 
            10.1038/s41467-021-27332-2"
COMMENT     Sequence Label: pAAV-hSyn-eLACCO1.1
FEATURES             Location/Qualifiers
     source          1..6602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..448
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     primer_bind     1409..1430
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1536..1557)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1797..1816)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2555..3143
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2608..2628)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3026..3037)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3175..3651
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3374..3393)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3691..3831
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3691..3820
                     /label=AAV2 ITR
     rep_origin      3906..4361
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3993..4012)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4203..4224
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4378..4397)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4497..4519
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4557..4575)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4643..4747
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4748..4816,4817..5608)
                     /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(4966..4985)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5779..6367
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6268..6287
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6429..6558
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
       61 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      121 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg
      181 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      241 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      301 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      361 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      421 aggagtcgtg tcgtgcctga gagcgcagtc gagaaggtac cggatcggac gaataacgcg
      481 tggatccacc ggtgagctcc cgcggcggcc gctctagatt agtctgcaga atcaccatgg
      541 gaatccaagg agggtctgtc ctgttcgggc tgctgctcgt cctggctgtc ttctgccatt
      601 caggtcatag cagatctatg ttctctccct tggcagtggc ccaggctcga cgctataggt
      661 ggaggattca aaccgcgtgg gatgctggaa cagtaggcta cagtctgttc caaaagttta
      721 cggagcgggt aaaggaactg acggatggac agcttgaggt ccagcctttt ccagcgggtg
      781 cagtcgtagg cacgtttgat atgttcgacg cggtcaagac tggagttttg gacggaatga
      841 atcctttcac gctgttttgg gcagggagga tgccggttac agctttcctc tccagctacg
      901 ccctcgggtt ggaccgacct gaccagtggg aaacatggtt ttactccctg ggtggcctcg
      961 acaacgcccg acgagccttt gccgagcaag gcctcttcta cgtgggacca gttcagcacg
     1021 acctcaatac aattcattca aggaagccga tccggagatt tgaggacttc aaaggtgtca
     1081 agctccgagt gccgggcggc atgattgcgg aagtttttgc ggccgcagga gcttccaccg
     1141 tcctcttgcc tgggggggag gtctaccccg ctctggaaag aggggtaata gattggagcc
     1201 acaacgtcta tatcatggcc gacaagcaga ggaacggcat caaggcgaac ttcgagatcc
     1261 gccacaacat cgaggacggc ggcgtgcagc tcgcctacca ctaccagcag aacaccccca
     1321 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag accaaacttt
     1381 cgaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     1441 ggatcactct cggcatggac gagctgtaca agggcggtac cggagggaga atggtgagca
     1501 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     1561 acggccacaa gttcagcgtg tccggcgagg gtgagggcga tgccacctcc ggcaagctga
     1621 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     1681 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     1741 tcttcaagtc ggccatgccc gaaggctaca tccaggagcg caccatcttc ttcaaggacg
     1801 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1861 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcat aagctggagt
     1921 acagctttaa cgatggtggt gcggctgatt ttgttgggcc tgcagtaaac tataacttgg
     1981 ggttccatca ggtggcaaag tacatcataa tggggcctcc agagacgcct gccattcatc
     2041 aacctgtaga cctcatggac ttcaccataa accttaatcg gtggcgcagc cttccaaagc
     2101 ctctgcaaga aagattcatc gcggcggttc acgagtactc ttggattcac tacgcaggca
     2161 ttcagaaggc taatttggag gcgtggccga aatatcggca ggcaggtgtt gaggtcatcc
     2221 ggctgagcaa cgaggatgtt agaaaattca ggcgactcgc tattcccatt tggtttaagt
     2281 gggctaagat ggataaatac tcacgagagg cattcgcgtc ccaactggag tatatgaaag
     2341 gtataggtta cgttacggac gaggaattga agggcttgag cctgggctcc acctccggct
     2401 ccggcaagcc cggctccggc gagggctcca ccaagggcga attccttgaa aatggtggga
     2461 catccttatc agagaaaaca gttcttctgc tggtgactcc atttctggca gcagcctgga
     2521 gccttcatcc ctaaggatca tcaagcttat cgataatcaa cctctggatt acaaaatttg
     2581 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     2641 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     2701 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     2761 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     2821 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     2881 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     2941 gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg
     3001 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     3061 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     3121 ctccctttgg gccgcctccc cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg
     3181 catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc
     3241 accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat
     3301 aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt
     3361 agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact
     3421 gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga
     3481 ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc
     3541 accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct
     3601 cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgattttg
     3661 taggtaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc
     3721 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     3781 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga
     3841 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc
     3901 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3961 gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct
     4021 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     4081 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     4141 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4201 tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga
     4261 tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa
     4321 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac
     4381 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     4441 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     4501 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     4561 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     4621 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     4681 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4741 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4801 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4861 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4921 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4981 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5041 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5101 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5161 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5221 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5281 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     5341 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     5401 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5461 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5521 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     5581 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     5641 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5701 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5761 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5821 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5881 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5941 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6001 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6061 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6121 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6181 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6241 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6301 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6361 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6421 tcacatgtcc tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc gggcgtcggg
     6481 cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact
     6541 ccatcactag gggttcctgc ggccgcacgc gtgtgtctag actgcagagg gccctgcgta
     6601 tg
//
