LOCUS       sequence-369499-        5127 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mitochondrial 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-cGFP (matrix)
FEATURES             Location/Qualifiers
     source          1..5127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        69..1251
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          300..1242
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1199..1219
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1289..1363
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     regulatory      1391..1400
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1397..1399,1400..1402,1403..1915)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIE"
     primer_bind     complement(1442..1463)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1703..1722)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2050..2071
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2182..2201)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2247..2302
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2301..2320)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2663..2679)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2663..2679)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2676..2698)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2687..2703
                     /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(2711..2717,2718..2735,2736..2741))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2756..2777
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2836..3031
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2882..3017
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3037..3171
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3074..3093)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3128..3147
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3239..3256)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3410..3998)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3490..3509)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4169..4960,4961..5029))
                     /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     4792..4811
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5030..5127,1..7))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttccgcgcac atttccccga aaagtgccac ctgggtcgac attgattatt gactagatca
       61 tcgcgtgagg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga
      121 agttgggggg aggggtcggc aattgaaccg gtgcctagag aaggtggcgc ggggtaaact
      181 gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata
      241 taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg
      301 taagtgccgt gtgtggttcc cgcgggcctg gcctctttac gggttatggc ccttgcgtgc
      361 cttgaattac ttccacgccc ctggctgcag tacgtgattc ttgatcccga gcttcgggtt
      421 ggaagtgggt gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga
      481 gttgaggcct ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc
      541 tgtctcgctg ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg
      601 ctttttttct ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg
      661 gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg
      721 cggggcctgc gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct
      781 gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc
      841 cggtcggcac cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc
      901 tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa
      961 agggcctttc cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc
     1021 aggcacctcg attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg
     1081 ttttatgcga tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg
     1141 cacttgatgt aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc
     1201 aagcctcaga cagtggttca aagttttttt cttccatttc aggtgtcgtg aggaattctg
     1261 cagtcgacgg taccgcgggc ccgccaccat gtccgtcctg acgccgctgc tgctgcgggg
     1321 cttgacaggc tcggcccggc ggctcccagt gccgcgcgcc aagatccatt cgttggggga
     1381 tccaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     1441 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     1501 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     1561 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     1621 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     1681 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     1741 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     1801 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     1861 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     1921 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     1981 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     2041 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggctaagg
     2101 ccgcactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct
     2161 ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg acatcatgaa
     2221 gccccttgag catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg
     2281 ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca
     2341 tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa
     2401 caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt
     2461 attccataga aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt
     2521 tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct
     2581 ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc tcgacctgca
     2641 gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     2701 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     2761 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     2821 cgtgccagcg gatccgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     2881 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     2941 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     3001 aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc agcttataat
     3061 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3121 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgctgcatt
     3181 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     3241 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3301 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3361 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3421 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3481 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3541 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3601 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3661 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3721 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3781 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3841 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3901 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3961 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4021 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4081 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4141 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4201 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4261 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     4321 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     4381 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     4441 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     4501 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     4561 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     4621 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     4681 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     4741 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     4801 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     4861 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4921 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     4981 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5041 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     5101 gtatttagaa aaataaacaa atagggg
//
