LOCUS       sequence-410574-        6573 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 6573)
  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 6573)
  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-HA-eLACCO2.1-NGR
FEATURES             Location/Qualifiers
     source          1..6573
                     /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     complement(1641..1662)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1902..1921)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2700..3288
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2753..2773)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3171..3182)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3320..3796
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3519..3538)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3836..3976
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3836..3965
                     /label=AAV2 ITR
     rep_origin      4051..4506
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4138..4157)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4348..4369
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4523..4542)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4642..4664
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4702..4720)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4788..4892
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4893..4961,4962..5753)
                     /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(5111..5130)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5924..6512
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6413..6432
                     /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 ccgccaccat gaagaccatc atcgccctgt cctacatctt ctgcctggtg ctgggcagat
      721 ctatgttctc tcccttggca gtggcccagg ctcgacgcta taggtggcgc attcaaaccg
      781 cgtgggatgc cggaacagta ggctacagtc tgttccaaaa gtttacggag cgggtaaagg
      841 aactgacgga tggacagctt gaggtccagc cttttccagc gagtgcagtc gtaggcacgt
      901 ttgatatgtt cgacgcggtc aagactggag ttttggacgg aatgaatcct ttcacgatct
      961 attgggcagg gaggatgccg gttacagctt tcctctccag ctacgccctc gggttggacc
     1021 gacctgacca gtgggaaaca tggttttact ccctgggtgg cctcgacaac gcccgacgag
     1081 cctttgccga gcaaggcctc ttctacgtgg gaccagttca gcacgacctc aatacaattc
     1141 attcaaagaa gccgatccgg agatttgagg acttcaaagg tgtcaagctc cgagtgccgg
     1201 gcggcatgat tgcggaagtt tttgcggccg caggagcttc caccgtcctc ttgcctgggg
     1261 gggaggtcaa ccccgctctg gaaagagggg taatagattg gagccacaac gtccatatca
     1321 tggccgacaa gcagaggaac ggcatcaagg cgaacttcga gatccgccac agcaccgagg
     1381 acggcggcgt gcagctcgcc taccactacc agcagaacac ccccatcggc gacggccccg
     1441 tgctgctgcc cgacaaccac tacctgagca cccagaccaa acttacgaaa gaccccaacg
     1501 agaagcgcga tcacatggtc ctgctggagt acgtgaccgc cgccgggatc actcgcggca
     1561 tggacgagct gtacaagggc ggtaccggag ggagaatggt gagcaagggc gaggagctgt
     1621 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     1681 gcgtgtccgg cgagggtgag ggcgatgcca cctccggcaa gctgtccctg aagttcatca
     1741 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     1801 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtcggcca
     1861 tgcccgaagg ctacatccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     1921 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     1981 tcgacttcaa ggaggacggc aacatcctgg ggcataagct ggagtacagc tttaacgatg
     2041 gtggtgcggc tgattttgtt gggcctgcag taaactataa cttggggttc catcaggtgg
     2101 caaagtacat cataatgggg cctccagaga cgcctgccat tcatcaacct gtagacctca
     2161 tggacttcac cataaacctt aatcggtggc gcagccttcc aaagcccctg caagaaagat
     2221 tcatcgcggc ggttcacgag tactcttgga ttcactacgc aggcattcag aaggctaatt
     2281 tggaggcgtg gccgaaatat cggcaggcag gtgttgaggt catccggctg agcaacgagg
     2341 atgttagaaa attcaggcga ctcgctattc ccatttggtt taagtgggct aagatggata
     2401 actactcacg agaggcattc gcgtctcaac tggagtatat gaaaggtata ggttacgtta
     2461 cggacgagga attgaagggc ttgggcctgg gctccacctc cggctccggc aagcccggct
     2521 ccggcgaggg ctccaccaag ggcgaattcg caggtagtgg tgcatctgga acaggagatg
     2581 cagaaggatc aggtgccctt cctgcccttg catgtagtct ggctccactg ggactggcat
     2641 tggttctgtg gacggtgttg ggcccctgtt aggcggccgc tcgaatcaag cttatcgata
     2701 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     2761 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     2821 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     2881 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     2941 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     3001 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3061 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     3121 cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     3181 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     3241 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag
     3301 cgctgctcga gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc
     3361 cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat
     3421 tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa
     3481 ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag
     3541 tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct
     3601 gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt
     3661 gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc
     3721 tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc
     3781 tcccttccct gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa
     3841 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     3901 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     3961 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     4021 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     4081 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc
     4141 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4201 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4261 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4321 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4381 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt
     4441 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     4501 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     4561 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     4621 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     4681 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     4741 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4801 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4861 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4921 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4981 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5041 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5101 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5161 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5221 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5281 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     5341 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5401 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     5461 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5521 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5581 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5641 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5701 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     5761 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5821 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5881 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5941 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6001 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6061 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6121 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6181 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6241 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6301 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6361 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6421 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6481 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6541 tcctggcctt ttgctggcct tttgctcaca tgt
//
