LOCUS       sequence-369496-        5127 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mitochondrial photoactivatable GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5127)
  AUTHORS   Vevea JD, Chapman ER
  TITLE     Mitofusin 2 Sustains the Axonal Mitochondrial Network to Support 
            Presynaptic Ca(2+) Homeostasis and the Synaptic Vesicle Cycle in Rat
            Hippocampal Axons.
  JOURNAL   J Neurosci. 2023 May 10;43(19):3421-3438. doi: 
            10.1523/JNEUROSCI.1356-22.2023. Epub 2023 Mar 30.
  PUBMED    36997314
REFERENCE   2  (bases 1 to 5127)
  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: 
            "10.1523/JNEUROSCI.1356-22.2023"; journalName: "J Neurosci"; date: 
            "2023-05-10- 10"; volume: "43"; issue: "19"; pages: "3421-3438"
COMMENT     Sequence Label: pEF mito-PAcGFP
FEATURES             Location/Qualifiers
     source          1..5127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        110..1292
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          341..1283
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1240..1260
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1330..1404
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     regulatory      1432..1441
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1438..1440,1441..1443,1444..1956)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     complement(1483..1504)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1744..1763)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2091..2112
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2223..2242)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2288..2343
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2342..2361)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2704..2720)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2704..2720)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2717..2739)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2728..2744
                     /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(2752..2758,2759..2776,2777..2782))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2797..2818
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2877..3072
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2923..3058
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3078..3212
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3115..3134)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3169..3188
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3280..3297)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3451..4039)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3531..3550)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4210..5001,5002..5070))
                     /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     4833..4852
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5071..5127,1..48))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
       61 aaaagtgcca cctgggtcga cattgattat tgactagatc atcgcgtgag gctccggtgc
      121 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
      181 caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
      241 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
      301 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc
      361 ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacgcc
      421 cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg tgggagagtt
      481 cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc
      541 gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct gctttcgata
      601 agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc tggcaagata
      661 gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg gccgcgggcg
      721 gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg cgagcgcggc
      781 caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg cctggcctcg
      841 cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca ccagttgcgt
      901 gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg aggacgcggc
      961 gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt ccgtcctcag
     1021 ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc gattagttct
     1081 cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg atggagtttc
     1141 cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg taattctcct
     1201 tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag acagtggttc
     1261 aaagtttttt tcttccattt caggtgtcgt gaggaattct gcagtcgacg gtaccgcggg
     1321 cccgccacca tgtccgtcct gacgccgctg ctgctgcggg gcttgacagg ctcggcccgg
     1381 cggctcccag tgccgcgcgc caagatccat tcgttggggg atccaccggt cgccaccatg
     1441 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     1501 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     1561 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     1621 gtgaccacct tcagctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     1681 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     1741 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     1801 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     1861 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     1921 atcaaggcca acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     1981 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     2041 ctgagccacc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     2101 ctggagttcg tgaccgccgc cgggatcact ctcggctaag gccgcactcc tcaggtgcag
     2161 gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga
     2221 gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact
     2281 tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc
     2341 tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt
     2401 tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga
     2461 ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg
     2521 acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta
     2581 aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc
     2641 atagctgtcc ctcttctctt atggagatcc ctcgacctgc agcccaagct tggcgtaatc
     2701 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     2761 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     2821 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatccgcat
     2881 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     2941 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3001 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3061 ggcttttgca aaaagctaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3121 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3181 aactcatcaa tgtatcttat catgtctgga tccgctgcat taatgaatcg gccaacgcgc
     3241 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3301 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3361 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3421 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3481 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3541 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3601 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3661 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3721 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3781 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3841 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3901 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3961 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4021 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4081 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4141 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4201 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4261 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4321 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4381 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4441 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4501 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4561 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4621 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4681 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4741 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4801 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4861 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4921 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4981 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5041 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5101 ttatcagggt tattgtctca tgagcgg
//
