LOCUS       sequence-329879-        5625 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of photoconvertible mEos2 targted to mitochondrial matrix
            to allow tracking of individual mitochondria and measurement of 
            fission-fusion rate.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5625)
  AUTHORS   Lewis TL Jr, Kwon SK, Lee A, Shaw R, Polleux F
  TITLE     MFF-dependent mitochondrial fission regulates presynaptic release 
            and axon branching by limiting axonal mitochondria size.
  JOURNAL   Nat Commun. 2018 Nov 27;9(1):5008. doi: 10.1038/s41467-018-07416-2.
  PUBMED    30479337
REFERENCE   2  (bases 1 to 5625)
  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: "2018-11-27"; volume: "9(1):5008. doi"; pages: " 
            10.1038/s41467-018-07416-2"
COMMENT     Sequence Label: pCAG mito-mEos2
FEATURES             Location/Qualifiers
     source          1..5625
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        18..397
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        399..676
                     /label=chicken beta-actin promoter
     intron          677..1693
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1617..1639
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1701..1720
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1754..1828
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             1862..2539
                     /codon_start=1
                     /product="monomeric variant of green-to-red photoswitchable
                     fluorescent protein EosFP (McKinney et al., 2009)"
                     /label=mEos2
                     /translation="SAIKPDMKIKLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVKE
                     GGPLPFAFDILTTAFHYGNRVFAKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICIARN
                     DITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIHMALLLE
                     GNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPDN
                     ARR"
     primer_bind     complement(2630..2649)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2695..2750
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2749..2768)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3141..3157)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3141..3157)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3154..3176)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3165..3181
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3189..3195,3196..3213,3214..3219))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3234..3255
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3314..3509
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3360..3495
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3515..3649
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3552..3571)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3606..3625
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3717..3734)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3888..4476)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3968..3987)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4647..5438,5439..5507))
                     /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     5270..5289
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5508..5612)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaagtgccac ctgggtcgac attgattatt gactagttat taatagtaat caattacggg
       61 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      121 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      181 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      241 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      301 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      361 gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc ccacgttctg
      421 cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta
      481 attattttgt gcagcgatgg gggcgggggg gggggggggg cgcgcgccrg gsggggsggg
      541 gsggggsgrg gggsggggsg gggsgaggcg gagaggtgcg gcggcagcca atcagagcgg
      601 cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg
      661 aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc
      721 cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg
      781 ggacggccct tctcctccgg gctgtaatta gcgcttggtt taatgacggc ttgtttcttt
      841 tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg gggggagcgg
      901 ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc
      961 cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga
     1021 ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg ggctgcgagg ggaacaaagg
     1081 ctgcgtgcgg ggtgtgtgcg tgggggggtg agcagggggt gtgggcgcgt cggtcgggct
     1141 gcaacccccc ctgcaccccc ctccccgagt tgctgagcac ggcccggctt cgggtgcggg
     1201 gctccgtacg gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg
     1261 ggtgccgggc ggggcggggc cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg
     1321 cccccggagc gccggcggct gtcgaggcgc ggcgagccgc agccattgcc ttttatggta
     1381 atcgtgcgag agggcgcagg gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg
     1441 gaggcgccgc cgcaccccct ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa
     1501 ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca
     1561 gcctcggggc tgtccgcggg gggacggctg ccttcggggg ggacggggca gggcggggtt
     1621 cggcttctgg cgtgtgaccg gcggctctag agcctctgct aaccatgttc atgccttctt
     1681 ctttttccta cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc attttggcaa
     1741 agaattgctc gagatgtccg tcctgacgcc gctgctgctg cggggcttga caggctcggc
     1801 ccggcggctc ccagtgccgc gcgccaagat ccattcgttg ggggatccac cggtcgccac
     1861 cagtgcgatt aagccagaca tgaagatcaa actccgtatg gaaggcaacg taaacgggca
     1921 ccactttgtg atcgacggag atggtacagg caagcctttt gagggaaaac agagtatgga
     1981 tcttgaagtc aaagagggcg gacctctgcc ttttgccttt gatatcctga ccactgcatt
     2041 ccattacggc aacagggtat tcgccaaata tccagacaac atacaagact attttaagca
     2101 gtcgtttcct aaggggtatt cgtgggaacg aagcttgact ttcgaagacg ggggcatttg
     2161 cattgccaga aacgacataa caatggaagg ggacactttc tataataaag ttcgatttta
     2221 tggtaccaac tttcccgcca atggtccagt tatgcagaag aagacgctga aatgggagcc
     2281 ctccactgag aaaatgtatg tgcgtgatgg agtgctgacg ggtgatattc atatggcttt
     2341 gttgcttgaa ggaaatgccc attaccgatg tgacttcaga actacttaca aagctaagga
     2401 gaagggtgtc aagttaccag gctaccactt tgtggaccac tgcattgaga ttttaagcca
     2461 tgacaaagat tacaacaagg ttaagctgta tgagcatgct gttgctcatt ctggattgcc
     2521 tgacaatgcc agacgataag cggccgcaat tcactcctca ggtgcaggct gcctatcaga
     2581 aggtggtggc tggtgtggcc aatgccctgg ctcacaaata ccactgagat ctttttccct
     2641 ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa
     2701 ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga
     2761 catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa
     2821 catatgccca tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata
     2881 tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga
     2941 ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac
     3001 atgttttact agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc
     3061 ttctcttatg gagatccctc gacctgcacc gtcgaccagc tggtcgacgg tgcaccgtcg
     3121 accagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     3181 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     3241 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     3301 tgccagcgga tccgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3361 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3421 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3481 gcttttttgg aggcctaggc ttttgcaaaa agctaacttg tttattgcag cttataatgg
     3541 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3601 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc gctgcattaa
     3661 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     3721 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3781 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3841 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3901 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3961 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4021 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4081 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4141 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4201 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4261 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4321 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4381 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4441 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4501 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4561 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4621 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4681 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4741 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4801 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4861 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4921 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4981 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5041 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5101 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5161 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5221 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5281 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5341 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5401 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5461 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5521 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5581 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccga
//
