LOCUS       sequence-352424-        6263 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the bacterial lactate 2-monooxygenase (LMO) enzyme in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6263)
  AUTHORS   Vaccari Cardoso B, Barrera I, Mosienko V, Gourine AV, Kasparov S, 
            Teschemacher AG
  TITLE     Expression of Microbial Enzymes in Mammalian Astrocytes to Modulate 
            Lactate Release.
  JOURNAL   Brain Sci. 2021 Aug 10;11(8). pii: brainsci11081056. doi: 
            10.3390/brainsci11081056.
  PUBMED    34439675
REFERENCE   2  (bases 1 to 6263)
  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: "Brain 
            Sci."; date: "2021-08-10"; volume: "11(8). pii: brainsci11081056. 
            doi"; pages: " 10.3390/brainsci11081056"
COMMENT     Sequence Label: pCMV-LMO-IRES-EGFP
FEATURES             Location/Qualifiers
     source          1..6263
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1594..1603
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1600..1602,1603..1605,1606..2319)
                     /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(1600..1602,1603..1605,1606..2316)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1645..1666)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1906..1925)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2253..2274
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2442..2563
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2479..2498)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2533..2552
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2570..3025)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2707..2728)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2919..2938
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3052..3156
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3158..3515
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3158..3349
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3181..3201)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3366..3501
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3550..4344
                     /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             3550..4344
                     /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(3604..3623)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4214..4233
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4532..4551)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4576..4623
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4952..5540
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5441..5460
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5715..6018
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6019..6222
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6173..6193
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gatcttcgaa gccgccacca tgagcaactg gggcgactac gagaacgaaa tctacggcca
       61 gggactcgtg ggagtcgctc ctacactgcc tatgagctac gccgactggg aagctcatgc
      121 tcagcaagca cttccacctg gcgtgctgtc ttatgtggct ggcggaagcg gagatgagca
      181 cacacagagg gctaacgtgg aagccttcaa gcactggggc ctgatgccta gaatgctgat
      241 ggctgccacc gagagggacc tgtctgttga gctgtggggc aagacatggg ccgctcctat
      301 gttcttcgct cccatcggag tgatcgccct gtgtgcccaa gatggacatg gcgacgctgc
      361 atctgctcag gcctctgcta gaaccggcgt gccctacatc acaagcacac tggctgtgtc
      421 cagcctcgag gacatcagaa agcacgctgg cgatacccct gcctacttcc agctgtacta
      481 ccccgaggat cgcgacctgg ccgagagctt tatcagaagg gctgaagagg ccggctacga
      541 cggcctggtt atcacactgg acacctggat cttcggctgg cggcctaggg atctgacaat
      601 cagcaacttc ccattcctga gaggcctgtg cctgaccaac tacgtgacag accccgtgtt
      661 ccagaagaag ttcaaggccc acagcggcgt ggaagctgag ggcctgagag ataaccctag
      721 actggccgcc gatttctggc acggcctgtt tggacactct gtgacctggg aagatattga
      781 ctgggtccga tccatcacca agatgcccgt gatcctgaag ggcatccagc atcctgacga
      841 tgctcggaga gctgtggata gcggagtgga cggcatctac tgttctaacc acggcggcag
      901 acaggccaat ggtggactgc ctgctctgga ctgtctgccc gaagtggtta aggccagcgg
      961 cgatacacct gtgctgttcg acagcggcat cagaacaggc gccgatgtgg tcaaggctct
     1021 ggctatggga gcttctgctg tcggcatcgg cagaccttat gcttggggag ctgctctcgg
     1081 cggctctaag ggaatcgagc atgtggctag aagcctgctg gccgaggccg atctgatcat
     1141 ggctgtggac ggctacagaa acctgaaaga gctgaccatc gacgccctga ggcctaccag
     1201 ataagatatc tgcagtcgac ggtaccagat gggggtcctg ggccccaggg tgtgcagcca
     1261 ctgacttggg gactgctggt ggggtaggga tgagggaggg aggggcattg tgatgtacag
     1321 ggctgctctg tgagatcaag ggtctcttaa gggtgggagc tggggcaggg actacgagag
     1381 cagccagatg ggctgaaagt ggaactcaag gggtttctgg cacctaccta cctgcttccc
     1441 gctggggggt ggggagttgg cccagagtct taagattggg gcagggtgga gaggtgggct
     1501 cttcctgctt cccactcatc ttatagcttt ctttccccag atccgaattc gagatccaaa
     1561 ccaagggtac cgcgggcccg ggatccaccg gtcgccacca tggtgagcaa gggcgaggag
     1621 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1681 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     1741 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     1801 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1861 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     1921 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1981 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     2041 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     2101 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     2161 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     2221 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     2281 gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgcga ctctagatca
     2341 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     2401 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     2461 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     2521 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc
     2581 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     2641 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     2701 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     2761 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     2821 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     2881 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     2941 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     3001 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     3061 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3121 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     3181 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3241 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3301 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3361 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3421 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     3481 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     3541 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3601 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3661 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3721 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3781 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3841 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     3901 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     3961 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4021 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     4081 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4141 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4201 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4261 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4321 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4381 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4441 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4501 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     4561 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     4621 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     4681 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     4741 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     4801 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     4861 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4921 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4981 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5041 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5101 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5161 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5221 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5281 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5341 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5401 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5461 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5521 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     5581 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     5641 ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     5701 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     5761 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     5821 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     5881 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     5941 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     6001 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     6061 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     6121 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     6181 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag
     6241 atccgctagc gctaccggac tca
//
