LOCUS       sequence-65245-e        7216 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  a nuclear-targeted, genetically-encoded sensor for lactate.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7216)
  AUTHORS   Barros LF, San Martin A, Sotelo-Hitschfeld T, Lerchundi R, 
            Fernandez-Moncada I, Ruminot I, Gutierrez R, Valdebenito R, Ceballo 
            S, Alegria K, Baeza-Lehnert F, Espinoza D
  TITLE     Small is fast: astrocytic glucose and lactate metabolism at cellular
            resolution.
  JOURNAL   Front Cell Neurosci. 2013;7:27. doi: 10.3389/fncel.2013.00027. Epub 
            2013 Mar 22.
  PUBMED    23526722
REFERENCE   2  (bases 1 to 7216)
  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: "Front Cell 
            Neurosci."; date: "2013"; volume: "7:27. doi"; pages: " 
            10.3389/fncel.2013.00027. Epub 2013 Mar 22"
COMMENT     Sequence Label: NLS-Laconic/pCMV-myc-nuc
FEATURES             Location/Qualifiers
     source          1..7216
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        12..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     544..564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      650..659
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             689..1396
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
     CDS             join(2198..2200,2201..2203,2204..2914)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2243..2264)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2504..2523)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2851..2872
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2927..2947
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2951..2971
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2975..2995
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3008..3037
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3073..3090)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3079..3303
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3349..3777
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3436..3455)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3646..3667
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3786..3806)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3791..4120
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3971..4106
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4033..4052
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4146..4940
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MVEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4200..4219)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4810..4829
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5114..5235
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5151..5170)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5205..5224
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5268..5285)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5439..6027)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5519..5538)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6198..6989,6990..7058))
                     /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     6821..6840
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7059..7163)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtaccgaatt cacattgatt attgagtagt tattaatagt aatcaattac ggggtcatta
       61 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      121 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      181 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      241 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      301 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      361 atctacggtt agtcatcgct attaccatag tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctttctggc
      601 taactagaga acccactgct tactggcacg tggaaattaa tacgacgtgg ccaccatggc
      661 ccaggtgcag ctgcaggtcg acctcgagat ggtgagcaag ggcgaggaga ccacaatggg
      721 cgtaatcaag cccgacatga agatcaagct gaagatggag ggcaacgtga atggccacgc
      781 cttcgtgatc gagggcgagg gcgagggcaa gccctacgac ggcaccaaca ccatcaacct
      841 ggaggtgaag gagggagccc ccctgccctt ctcctacgac attctgacca ccgcgttcgc
      901 ctacggcaac agggccttca ccaagtaccc cgacgacatc cccaactact tcaagcagtc
      961 cttccccgag ggctactctt gggagcgcac catgaccttc gaggacaagg gcatcgtgaa
     1021 ggtgaagtcc gacatctcca tggaggagga ctccttcatc tacgagatac acctcaaggg
     1081 cgagaacttc ccccccaacg gccccgtgat gcagaagaag accaccggct gggacgcctc
     1141 caccgagagg atgtacgtgc gcgacggcgt gctgaagggc gacgtcaagc acaagctgct
     1201 gctggagggc ggcggccacc accgcgttga cttcaagacc atctacaggg ccaagaaggc
     1261 ggtgaagctg cccgactatc actttgtgga ccaccgcatc gagatcctga accacgacaa
     1321 ggactacaac aaggtgaccg tttacgagag cgccgtggcc cgcaactcca ccgacggcat
     1381 ggacgagctg tacaagagat ctggtaccat gattgtttta cccagacgcc tgtcagacga
     1441 ggttgccgat cgtgtgcggg cgctgattga tgaaaaaaac ctggaagcgg gcatgaagtt
     1501 gcccgctgag cgccaactgg cgatgcaact cggcgtatca cgtaattcac tgcgcgaggc
     1561 gctggcaaaa ctggtgagtg aaggcgtgct gctcagtcga cgcggcggcg ggacgtttat
     1621 tcgctggcgt catgacacat ggtcggagca aaacatcgtc cagccgctaa aaacactgat
     1681 ggccgatgat ccggattaca gtttcgatat tctggaagcc cgctacgcca ttgaagccag
     1741 caccgcatgg catgcggcaa tgcgcgccac acctggcgac aaagaaaaga ttcagctttg
     1801 ctttgaagca acgctaagtg aagacccgga tatcgcctca caagcggacg ttcgttttca
     1861 tctggcgatt gccgaagcct cacataacat cgtgctgctg caaaccatgc gcggtttctt
     1921 cgatgtcctg caatcctcag tgaagcatag ccgtcagcgg atgtatctgg tgccaccggt
     1981 tttttcacaa ctgaccgaac aacatcaggc tgtcattgac gccatttttg ccggtgatgc
     2041 tgacggggcg cgtaaagcaa tgatggcgca ccttagtttt gttcacacca ccatgaaacg
     2101 attcgatgaa gatcaggctc gccacgcacg gattacccgc ctgcccggtg agcataatga
     2161 gcattcgagg gagaaaaacg cacttaagag atctaccatg gtgagcaagg gcgaggagct
     2221 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     2281 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat
     2341 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg
     2401 cctgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     2461 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     2521 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     2581 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     2641 ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat
     2701 ccgccacaac atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc
     2761 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccgccct
     2821 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     2881 cgggatcact ctcggcatgg acgagctgta caaggcggcc gcagatccaa aaaagaagag
     2941 aaaggtagat ccaaaaaaga agagaaaggt agatccaaaa aagaagagaa aggtagatac
     3001 ggccgcagaa caaaaactca tctcagaaga ggatctgaat ggggccgcat agtctagaag
     3061 ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     3121 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     3181 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     3241 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatggc ccgggctcta
     3301 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     3361 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3421 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3481 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3541 acctctcccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     3601 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3661 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     3721 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     3781 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3841 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3901 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3961 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc ccctaggctg
     4021 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     4081 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctccccc cccgggaggt
     4141 ccacaatggt tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4201 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4261 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4321 aactccagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4381 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4441 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4501 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4561 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4621 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgtatgc
     4681 ccgacggcga ggatctcgtc gtgactcatg gcgatgcctg cttgccgaat atcatggtgg
     4741 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4801 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4861 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4921 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     4981 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5041 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5101 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5161 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5221 catcaatgta tcttatcatg tctgtatacc gtcgatcttt ccgcttcctc gctcactgac
     5281 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5341 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5401 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5461 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5521 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5581 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5641 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tatgcacgaa
     5701 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5761 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5821 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     5881 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5941 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6001 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6061 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6121 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6181 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6241 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6301 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6361 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6421 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6481 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6541 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6601 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6661 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6721 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6781 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6841 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6901 ttaccgctgt tgagatccag ttcgatgtaa cccactcggg cacccaactg atcttcagca
     6961 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7021 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7081 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7141 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcagatcg
     7201 acggatcggg agatcg
//
