LOCUS       sequence-329820-        6203 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the bacterial lactate oxidase (LOx) enzyme in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6203)
  AUTHORS   Vaccari Cardoso B, Shevelkin AV, Terrillion C, Mychko O, Mosienko V,
            Kasparov S, Pletnikov MV, Teschemacher AG
  TITLE     Reducing l-lactate release from hippocampal astrocytes by 
            intracellular oxidation increases novelty induced activity in mice.
  JOURNAL   Glia. 2021 Jan 5. doi: 10.1002/glia.23960.
  PUBMED    33400321
REFERENCE   2  (bases 1 to 6203)
  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: "Glia. 2021 
            Jan 5. doi: 10.1002/glia.23960."
COMMENT     Sequence Label: pCMV-LOx-IRES-EGFP
FEATURES             Location/Qualifiers
     source          1..6203
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      398..407
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(404..406,407..409,410..1123)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(404..406,407..409,410..1120)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(449..470)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(710..729)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1057..1078
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1246..1367
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1283..1302)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1337..1356
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1374..1829)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1511..1532)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1723..1742
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1856..1960
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1962..2319
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1962..2153
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1985..2005)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2170..2305
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2354..3148
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2354..3148
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2408..2427)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3018..3037
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3336..3355)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3380..3427
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3756..4344
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4245..4264
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4519..4822
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4823..5026
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4977..4997
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 agtactgaga tatctgcagt cgacggtacc agatgggggt cctgggcccc agggtgtgca
       61 gccactgact tggggactgc tggtggggta gggatgaggg agggaggggc attgtgatgt
      121 acagggctgc tctgtgagat caagggtctc ttaagggtgg gagctggggc agggactacg
      181 agagcagcca gatgggctga aagtggaact caaggggttt ctggcaccta cctacctgct
      241 tcccgctggg gggtggggag ttggcccaga gtcttaagat tggggcaggg tggagaggtg
      301 ggctcttcct gcttcccact catcttatag ctttctttcc ccagatccga attcgagatc
      361 caaaccaagg gtaccgcggg cccgggatcc accggtcgcc accatggtga gcaagggcga
      421 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
      481 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
      541 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
      601 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
      661 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
      721 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
      781 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
      841 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
      901 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
      961 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     1021 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     1081 cgccgccggg atcactctcg gcatggacga gctgtacaag taaagcggcc gcgactctag
     1141 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     1201 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     1261 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     1321 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt
     1381 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     1441 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     1501 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     1561 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     1621 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     1681 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     1741 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     1801 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     1861 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     1921 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     1981 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     2041 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2101 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2161 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2221 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     2281 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     2341 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     2401 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     2461 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     2521 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     2581 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     2641 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     2701 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     2761 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     2821 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     2881 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     2941 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     3001 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     3061 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     3121 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     3181 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     3241 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     3301 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     3361 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     3421 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     3481 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     3541 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     3601 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     3661 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     3721 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3781 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3841 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3901 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3961 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4021 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4081 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4141 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4201 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4261 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4321 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4381 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     4441 cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag
     4501 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     4561 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     4621 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     4681 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     4741 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     4801 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     4861 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     4921 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     4981 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
     5041 tcagatccgc tagcgctacc ggactcagat cttcgaagcc gccaccatga acaacaacga
     5101 catcgagtac aacgccccta gcgagatcaa gtacatcgac gtggtcaaca cctacgacct
     5161 ggaagaggaa gccagcaagg ttgtccctca cggcggcttc aactatatcg ctggcgctag
     5221 cggagatgag tggaccaaga gagccaacga cagagcctgg aagcacaagc tgctgtaccc
     5281 tagactggcc caggatgtgg aagcccctga cacctctaca gagatcctgg gacacaagat
     5341 caaggcccct ttcatcatgg cccctatcgc cgctcatgga ctggctcaca caacaaaaga
     5401 ggccggcaca gccagagccg tgtctgagtt tggcaccatc atgagcatca gcgcctacag
     5461 cggcgccacc ttcgaggaaa tctctgaggg actgaacggc ggacccagat ggttccaaat
     5521 ctacatggcc aaggacgacc agcagaacag agacatcctg gacgaggcca agtctgatgg
     5581 cgccacagcc atcatcctga cagccgactc taccgtgtcc ggcaacagag atagggacgt
     5641 gaagaacaag ttcgtgtacc ccttcggcat gcccatcgtg cagagatacc tgagaggcac
     5701 agccgagggc atgagcctga acaacatcta cggcgccagc aagcagaaga tcagccccag
     5761 ggacatcgaa gagatcgccg gacattctgg cctgcctgtg ttcgtgaagg gcatccagca
     5821 tcctgaggac gccgacatgg ctatcaagag aggcgcttcc ggaatctggg tgtccaacca
     5881 tggtgccaga cagctgtacg aggcccctgg ctctttcgac acactgcctg ctatcgccga
     5941 gcgcgtgaac aagagggtgc caatcgtgtt cgatagcggc gtgcggagag gcgagcatgt
     6001 ggctaaagct cttgcctctg gcgccgatgt tgtggctctt ggtagacccg tgctgttcgg
     6061 actggcactt ggaggatggc agggcgctta cagcgtgctg gactacttcc agaaagacct
     6121 gaccagagtg atgcagctga ccggctctca gaacgtcgag gacctgaaag gcctggacct
     6181 gttcgacaac ccctacggct acg
//
