LOCUS       sequence-453641-        6731 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To explore mitochondrial lactate dynamics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6731)
  AUTHORS   Rauseo D, Contreras-Baeza Y, Faurand H, Carcamo N, Suarez R, von 
            Faber-Castell A, Silva F, Mora-Gonzalez V, Wyss MT, Baeza-Lehnert F,
            Ruminot I, Alvarez-Navarro C, Martin AS, Weber B, Sandoval PY, 
            Barros LF
  TITLE     Lactate-carried Mitochondrial Energy Overflow
  JOURNAL   bioRxiv 2024.07.19.604361
REFERENCE   2  (bases 1 to 6731)
  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: "bioRxiv 
            2024.07.19.604361"
COMMENT     Sequence Label: Mito-CanlonicSF/pCMV-myc-mito
FEATURES             Location/Qualifiers
     source          1..6731
                     /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"
     CDS             636..710
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             1284..1535
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGPVLLPDNH
                     YLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     1472..1493
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1599..1620)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1860..1879)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2523..2552
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2588..2605)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2594..2818
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2864..3292
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2951..2970)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3161..3182
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3301..3321)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3306..3635
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3486..3621
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3548..3567
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3661..4455
                     /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(3715..3734)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4325..4344
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4629..4750
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4666..4685)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4720..4739
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4783..4800)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4954..5542)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5034..5053)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5713..6504,6505..6573))
                     /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     6336..6355
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6574..6678)
                     /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 tgaccatgtc cgtcctgacg ccgctgctgc
      661 tgcggggctt gacaggctcg gcccggcggc tcccagtgcc gcgcgccaag atccattcgt
      721 tgctgcagat gttttctcct ctggcggtgg cccaagcgag gcgctaccgt tggcgcatcc
      781 agaccgcttg ggatgcgggc acggtaggct attccctgtt ccagaagttc accgagcgtg
      841 ttaaggagct caccgacggc cagttggagg ttcagccctt ccccgcaggg gccgtggtgg
      901 gcacctttga catgtttgac gcggtgaaga cgggcgtttt ggacgggatg aaccccttca
      961 ccctcgcatg ggcggggcgg atgccggtga cggcctttct ttcctcctat gccctgggtt
     1021 tggaccggcc cgaccagtgg gagacctggt tctacagcct gggcggcctg gacatcgccc
     1081 ggagggcctt cgccgagcag gggcttttct acgtgggccc cgtgcaacac gatttgaaca
     1141 tcatccactc caaaaagccg atccgtcgct ttgaggattt caaaggggta aagctgcggg
     1201 tgccgggcgg aatgattgct gaagtctttg ccgccgcggg ggcctccacg gtccttctac
     1261 tagaaaacgt ctatatcaag gccgacaagc agaagaacgg catcaaggcg aacttcaaga
     1321 tccgccacaa catcgaggac ggcggcgtgc agctcgccta ccactaccag cagaacaccc
     1381 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcgtc cagtccaagc
     1441 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1501 ccgggatcac tctcggcatg gacgagctgt acaagggtgg taccggtgga tctatggtga
     1561 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     1621 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     1681 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     1741 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     1801 acttcttcaa gtccgccatg cccgaaggct acatccagga gcgcaccatc ttcttcaagg
     1861 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     1921 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     1981 agtacaacac cagacccgga ggcgaggtct atcctgcctt ggagcgtggg gtcattgatg
     2041 cggccgactt tgtggggcct gcggtcaact acaatctggg cttccaccag gtggccaagt
     2101 acatcattat ggggcccccg gagaccccgg ccatccatca gccggtggac cttatggact
     2161 tcaccatcaa cctcaatcgg tggcggtcgc tacccaagcc tttgcaagag cgtttcatcg
     2221 ccgctgttca cgaatacagt tggattcact acgccgggat tcaaaaggcc aacctcgagg
     2281 cctggcccaa gtaccgccag gcgggtgtgg aggtcatccg cctgtctaac gaggacgtgc
     2341 gcaagttccg ccgtctggcc atccccatct ggttcaagtg ggccaagatg gacaagtact
     2401 cgcgggaggc cttcgccagc cagctggagt acatgaaggg catcggctac gttacggacg
     2461 aggagctcaa ggggctttcc ctttagctgc aggtcgacct cgagatcaaa cgggcggccg
     2521 cagaacaaaa actcatctca gaagaggatc tgaatggggc cgcatagtct agaagctcgc
     2581 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     2641 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     2701 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     2761 aagggggagg attgggaaga caatagcagg catgctgggg atggcccggg ctctatggct
     2821 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     2881 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     2941 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3001 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3061 tcccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3121 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3181 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3241 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3301 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3361 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3421 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3481 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccta ggctgactaa
     3541 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     3601 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct ccccccccgg gaggtccaca
     3661 atggttgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     3721 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     3781 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactc
     3841 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     3901 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     3961 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4021 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4081 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4141 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg tatgcccgac
     4201 ggcgaggatc tcgtcgtgac tcatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4261 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4321 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4381 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4441 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     4501 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4561 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4621 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4681 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4741 atgtatctta tcatgtctgt ataccgtcga tctttccgct tcctcgctca ctgactcgct
     4801 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4861 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4921 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4981 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5041 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5101 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5161 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtatgc acgaaccccc
     5221 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5281 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5341 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5401 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5461 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5521 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5581 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5641 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5701 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5761 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5821 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5881 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5941 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6001 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6061 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6121 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6181 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6241 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6301 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6361 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6421 gctgttgaga tccagttcga tgtaacccac tcgggcaccc aactgatctt cagcatcttt
     6481 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6541 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6601 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6661 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtca gatcgacgga
     6721 tcgggagatc g
//
