LOCUS       sequence-329929-        5630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mito-EYFP (mitochondrial matrix targeted EYFP) for 
            visualization of mitochondrial dynamics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5630)
  AUTHORS   Lewis TL Jr, Turi GF, Kwon SK, Losonczy A, Polleux F
  TITLE     Progressive Decrease of Mitochondrial Motility during Maturation of 
            Cortical Axons In Vitro and In Vivo.
  JOURNAL   Curr Biol. 2016 Oct 10;26(19):2602-2608. doi: 
            10.1016/j.cub.2016.07.064. Epub 2016 Sep 15.
  PUBMED    27641765
REFERENCE   2  (bases 1 to 5630)
  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"; doi: 
            "10.1016/j.cub.2016.07.064"; journalName: "Curr Biol"; date: 
            "2016-10-10- 10"; volume: "26"; issue: "19"; pages: "2602-2608"
COMMENT     Sequence Label: pCAG mitoYFP
FEATURES             Location/Qualifiers
     source          1..5630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(45..633)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(125..144)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(804..1595,1596..1664))
                     /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     1427..1446
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1665..1769)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        1800..2179
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          2421..3437
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3361..3383
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3445..3464
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3498..3572
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     regulatory      3600..3609
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3606..3608,3609..3611,3612..4325)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3651..3672)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3912..3931)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4259..4280
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4417..4436)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4482..4537
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4536..4555)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4928..4944)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4928..4944)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4941..4963)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4952..4968
                     /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(4976..4982,4983..5000,5001..5006))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5021..5042
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5101..5296
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5147..5282
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5302..5436
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5339..5358)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5393..5412
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5504..5521)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
       61 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
      121 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
      181 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
      241 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
      301 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
      361 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
      421 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
      481 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
      541 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
      601 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
      661 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
      721 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
      781 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
      841 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
      901 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
      961 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     1021 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     1081 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     1141 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     1201 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     1261 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     1321 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     1381 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     1441 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     1501 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     1561 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     1621 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     1681 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     1741 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgggtcg
     1801 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     1861 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     1921 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     1981 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     2041 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     2101 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     2161 agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc
     2221 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
     2281 gggggcgggg ggrggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga
     2341 gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa
     2401 agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg
     2461 ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg
     2521 ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga cggcttgttt
     2581 cttttctgtg gctgcgtgaa agccttgagg ggctccggga gggccctttg tgcgggggga
     2641 gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc
     2701 tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc
     2761 gcgaggggag cgcggccggg ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca
     2821 aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg
     2881 ggctgcaacc ccccctgcac ccccctcccc gagttgctga gcacggcccg gcttcgggtg
     2941 cggggctccg tacggggcgt ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg
     3001 tgggggtgcc gggcggggcg gggccgcctc gggccgggga gggctcgggg gaggggcgcg
     3061 gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat tgccttttat
     3121 ggtaatcgtg cgagagggcg cagggacttc ctttgtccca aatctgtgcg gagccgaaat
     3181 ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca
     3241 ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc
     3301 tccagcctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg ggcagggcgg
     3361 ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat gttcatgcct
     3421 tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg
     3481 gcaaagaatt gctcgagatg tccgtcctga cgccgctgct gctgcggggc ttgacaggct
     3541 cggcccggcg gctcccagtg ccgcgcgcca agatccattc gttgggggat ccaccggtcg
     3601 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     3661 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     3721 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     3781 ccaccctcgt gaccaccttc ggctacggcc tgcagtgctt cgcccgctac cccgaccaca
     3841 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     3901 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     3961 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     4021 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     4081 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     4141 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     4201 accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     4261 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     4321 agtaaagcgg ccgcaattca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg
     4381 tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta
     4441 tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt
     4501 cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc
     4561 aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat
     4621 gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc
     4681 tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt
     4741 ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc
     4801 cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag
     4861 atccctcgac ctgcaccgtc gaccagctgg tcgacggtgc accgtcgacc agcttggcgt
     4921 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4981 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5041 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc
     5101 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     5161 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     5221 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     5281 cctaggcttt tgcaaaaagc taacttgttt attgcagctt ataatggtta caaataaagc
     5341 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5401 tccaaactca tcaatgtatc ttatcatgtc tggatccgct gcattaatga atcggccaac
     5461 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5521 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5581 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
//
