LOCUS       sequence-319272-        6362 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ubiquitous expression of mtON in C. elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6362)
  AUTHORS   Berry BJ, Trewin AJ, Milliken AS, Baldzizhar A, Amitrano AM, Lim Y, 
            Kim M, Wojtovich AP
  TITLE     Optogenetic control of mitochondrial protonmotive force to impact 
            cellular stress resistance.
  JOURNAL   EMBO Rep. 2020 Apr 3;21(4):e49113. doi: 10.15252/embr.201949113. 
            Epub 2020 Feb 11.
  PUBMED    32043300
REFERENCE   2  (bases 1 to 6362)
  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: "EMBO Rep.";
            date: "2020-04-3"; volume: "21(4):e49113. doi"; pages: " 
            10.15252/embr.201949113. Epub 2020 Feb 11"
COMMENT     Sequence Label: pBJB20 - eft-3p::mitofilin::Mac::GFP::unc-54 3...
FEATURES             Location/Qualifiers
     source          1..6362
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..600
                     /gene="eft-3"
                     /label=eft-3 promoter
                     /note="promoter for the C. elegans elongation factor 
                     1-alpha gene (Mitrovich and Anderson, 2000)"
     CDS             join(2143..2145,2146..2859)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2143..2145,2146..2856)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2185..2206)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2446..2465)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2793..2814
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3782..3804)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3904..3923
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4001..4019)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4087..4191
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4192..4260,4261..5052)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGEEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4410..4429)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5223..5811
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5712..5731
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5965..5982
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6099..6120
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6135..6140,6141..6158,6159..6165)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6173..6189
                     /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)."
     primer_bind     6178..6200
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6197..6213
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6197..6213
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gcacctttgg tcttttattg tcaacttcca ttggttcttc cattgtttct gttaaattaa
       61 tgaatttttc ataaaataaa gacattatac aatataaaaa tgaagaattt attgaaaata
      121 aactgccaga gagaaaaagt atgcaacact cccgccgaga gtgtttgaaa tggtgtacgg
      181 tacattttcg tgctaggagt tagatgtgca ggcagcaacg agagggggag agattttttt
      241 gggccttgtg aaattaacgt gagttttctg gtcatctgac taatcatgtt ggttttttgt
      301 tggtttattt tgtttttatc tttgttttta tccagattag gaaatttaaa ttttatgaat
      361 ttataatgag gtcaaacatt cagtcccagc gtttttcctg ttctcactgt ttagtcgaat
      421 ttttatttta ggctttcaac aaatgttcta actgtcttat ttgtgacctc actttttata
      481 tttttttaat ttttaaaaat attagaagtt tctaggataa ttttttcgac ttttattctc
      541 tctaccgtcc gcactcttct tacttttaaa ttaaattgtt tttttttcag ttgggaaaca
      601 ctttgctctc tagaaaaaat gctgcgggcc tgtcagttat ccggcgtgac cgtcgcggcc
      661 cagagttgtc tgtgtgggaa gttcgtcctc cgtccactac gaccatgccg tagatactcc
      721 acttcaagca gttctgggtt gactgctggt aagattgctg gagctggcct tttgttcgtt
      781 ggtggaggga ttggtggcac tatcctgtat gccaaatggg attcccattt ccgggaaagt
      841 gtagagaaaa ccatccctta ctcagacaaa ctctttggga tggttcttgg ttctgcgcct
      901 tatactgttc cattaccaaa gaaaccggtt cagtctggtc cactaaaaat ctccagtgta
      961 tcagaggtga tgaaagactc caaactgcct gtggcacaga gccagaaaac gaagggagac
     1021 actccagctt cagcagcatc tacaggggca gctcaaatta tttctgcagc aggtgataca
     1081 ctttcggtcc cggcccctgc agttcagcat gaggacacta taaaaaccga atgtcccaac
     1141 actaatgaag gaaaatctac gtctgaaacc acagaggaac ctgcaggctt gatcgtggac
     1201 cagttcgagg aggtgctgat gaagaccagc cagctgttcc cactgccaac cgctacccag
     1261 agcgcccagc caacccacgt ggcccccgtg ccaaccgtgc tgcccgacac ccccatctac
     1321 gagaccgtgg gcgacagcgg cagcaagacc ctgtgggtgg tgttcgtgct gatgctgatc
     1381 gccagcgccg ccttcaccgc cctgagctgg aagatccccg tgaacaggag gctgtaccac
     1441 gtgatcacca ccatcatcac cctgaccgcc gccctgagct acttcgctat ggctaccggc
     1501 cacggagtgg ccctgaacaa gatcgtgatc aggacccagc acgaccacgt gcccgacacc
     1561 tacgagaccg tgtaccgaca ggtgtactac gccaggtaca tcgactgggc tatcaccacc
     1621 ccactgctgc tgctggacct gggactgctg gctggaatga gcggagccca catcttcatg
     1681 gccatcgtgg ctgacctgat catggtgctg accggcctgt tcgctgcttt cggcagcgag
     1741 ggaaccccac agaagtgggg atggtacacc atcgcctgca tcgcctacat cttcgtggtg
     1801 tggcacctgg tgctgaacgg cggcgccaac gccagggtga agggcgagaa gctgaggagc
     1861 ttcttcgtgg ccatcggagc ttacaccctg atcctgtgga ccgcttaccc aatcgtgtgg
     1921 ggactggctg acggagctag gaagatcgga gtggacggag agatcatcgc ttacgctgtg
     1981 ctggacgtgc tggctaaggg agtgttcgga gcttggctgc tggtgaccca cgccaacctg
     2041 agggagagcg acgtggagct gaacggcttc tgggccaacg gcctgaacag ggagggcgcc
     2101 atcaggatcg gcgaggacga cggcgcccga ccggtagtag cagtgagcaa gggcgaggag
     2161 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     2221 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     2281 atttgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     2341 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     2401 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     2461 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     2521 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     2581 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     2641 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     2701 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     2761 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     2821 gccgggatca ctctcggcat ggacgagctg tacaagtaag cggccgcaat tccaactgag
     2881 cgccggtcgc taccattacc aacttgtctg gtgtcaaaaa taataggggc cgctgtcatc
     2941 agagtaagtt taaactgagt tctactaact aacgagtaat atttaaattt tcagcatctc
     3001 gcgcccgtgc ctctgacttc taagtccaat tactcttcaa catccctaca tgctctttct
     3061 ccctgtgctc ccacccccta tttttgttat tatcaaaaaa cttctcttaa tttctttgtt
     3121 ttttagcttc ttttaagtca cctctaacaa tgaaattgtg tagattcaaa aatagaatta
     3181 attcgtaata aaaagtcgaa aaaaattgtg ctccctcccc ccattaataa taattctatc
     3241 ccaaaatcta cacaatgttc tgtgtacact tcttatgttt tttacttctg ataaattttt
     3301 ttgaaacatc atagaaaaaa ccgcacacaa aataccttat catatgttac gtttcagttt
     3361 atgaccgcaa tttttatttc ttcgcacgtc tgggcctctc atgacgtcaa atcatgctca
     3421 tcgtgaaaaa gttttggagt atttttggaa tttttcaatc aagtgaaagt ttatgaaatt
     3481 aattttcctg cttttgcttt ttggggtttc ccctattgtt tgtcaagatt tcgaggacgg
     3541 cgtttttctt gctaaaatca caagtattga tgagcacgat gcaagaaaga tcggaagaag
     3601 gtttgggttt gaggctcagt ggaaggtgag tagaagttga taatttgaaa gtggagtagt
     3661 gtctatgggg tttttgcctt aaatgacaga atacattccc aatataccaa acataactgt
     3721 ttcctactag tcggccgtac gggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     3781 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     3841 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     3901 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     3961 gatgcgtaag gagaaaatac cgcatcaggc ggccttaagg gcctcgtgat acgcctattt
     4021 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     4081 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4141 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     4201 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     4261 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     4321 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     4381 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     4441 gccggtgaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     4501 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     4561 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     4621 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     4681 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     4741 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4801 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4861 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4921 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4981 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     5041 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     5101 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5161 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5221 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5281 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5341 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     5401 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5461 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5521 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5581 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5641 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5701 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5761 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5821 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5881 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     5941 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     6001 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     6061 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     6121 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     6181 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct gtaagtttaa
     6241 acatgatctt actaactaac tattctcatt taaattttca gagcttaaaa atggctgaaa
     6301 tcactcacaa cgatggatac gctaacaact tggaaatgaa ataagctggc cggccaaagc
     6361 tt
//
