LOCUS       sequence-306913-        6367 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 6367)
  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 6367)
  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..6367
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        16..615
                     /gene="eft-3"
                     /label=eft-3 promoter
                     /note="promoter for the C. elegans elongation factor 
                     1-alpha gene (Mitrovich and Anderson, 2000)"
     CDS             join(2158..2160,2161..2874)
                     /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(2158..2160,2161..2871)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2200..2221)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2461..2480)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2808..2829
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3802..3824)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3924..3943
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4021..4039)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4107..4211
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4212..4280,4281..5072)
                     /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(4430..4449)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5243..5831
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5732..5751
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5985..6002
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6119..6140
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6155..6160,6161..6178,6179..6185)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6193..6209
                     /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     6198..6220
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6217..6233
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6217..6233
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ggccggccaa agcttgcacc tttggtcttt tattgtcaac ttccattggt tcttccattg
       61 tttctgttaa attaatgaat ttttcataaa ataaagacat tatacaatat aaaaatgaag
      121 aatttattga aaataaactg ccagagagaa aaagtatgca acactcccgc cgagagtgtt
      181 tgaaatggtg tacggtacat tttcgtgcta ggagttagat gtgcaggcag caacgagagg
      241 gggagagatt tttttgggcc ttgtgaaatt aacgtgagtt ttctggtcat ctgactaatc
      301 atgttggttt tttgttggtt tattttgttt ttatctttgt ttttatccag attaggaaat
      361 ttaaatttta tgaatttata atgaggtcaa acattcagtc ccagcgtttt tcctgttctc
      421 actgtttagt cgaattttta ttttaggctt tcaacaaatg ttctaactgt cttatttgtg
      481 acctcacttt ttatattttt ttaattttta aaaatattag aagtttctag gataattttt
      541 tcgactttta ttctctctac cgtccgcact cttcttactt ttaaattaaa ttgttttttt
      601 ttcagttggg aaacactttg ctctctagaa aaaatgctgc gggcctgtca gttatccggc
      661 gtgaccgtcg cggcccagag ttgtctgtgt gggaagttcg tcctccgtcc actacgacca
      721 tgccgtagat actccacttc aagcagttct gggttgactg ctggtaagat tgctggagct
      781 ggccttttgt tcgttggtgg agggattggt ggcactatcc tgtatgccaa atgggattcc
      841 catttccggg aaagtgtaga gaaaaccatc ccttactcag acaaactctt tgggatggtt
      901 cttggttctg cgccttatac tgttccatta ccaaagaaac cggttcagtc tggtccacta
      961 aaaatctcca gtgtatcaga ggtgatgaaa gactccaaac tgcctgtggc acagagccag
     1021 aaaacgaagg gagacactcc agcttcagca gcatctacag gggcagctca aattatttct
     1081 gcagcaggtg atacactttc ggtcccggcc cctgcagttc agcatgagga cactataaaa
     1141 accgaatgtc ccaacactaa tgaaggaaaa tctacgtctg aaaccacaga ggaacctgca
     1201 ggcttgatcg tggaccagtt cgaggaggtg ctgatgaaga ccagccagct gttcccactg
     1261 ccaaccgcta cccagagcgc ccagccaacc cacgtggccc ccgtgccaac cgtgctgccc
     1321 gacaccccca tctacgagac cgtgggcgac agcggcagca agaccctgtg ggtggtgttc
     1381 gtgctgatgc tgatcgccag cgccgccttc accgccctga gctggaagat ccccgtgaac
     1441 aggaggctgt accacgtgat caccaccatc atcaccctga ccgccgccct gagctacttc
     1501 gctatggcta ccggccacgg agtggccctg aacaagatcg tgatcaggac ccagcacgac
     1561 cacgtgcccg acacctacga gaccgtgtac cgacaggtgt actacgccag gtacatcgac
     1621 tgggctatca ccaccccact gctgctgctg gacctgggac tgctggctgg aatgagcgga
     1681 gcccacatct tcatggccat cgtggctgac ctgatcatgg tgctgaccgg cctgttcgct
     1741 gctttcggca gcgagggaac cccacagaag tggggatggt acaccatcgc ctgcatcgcc
     1801 tacatcttcg tggtgtggca cctggtgctg aacggcggcg ccaacgccag ggtgaagggc
     1861 gagaagctga ggagcttctt cgtggccatc ggagcttaca ccctgatcct gtggaccgct
     1921 tacccaatcg tgtggggact ggctgacgga gctaggaaga tcggagtgga cggagagatc
     1981 atcgcttacg ctgtgctgga cgtgctggct aagggagtgt tcggagcttg gctgctggtg
     2041 acccacgcca acctgaggga gagcgacgtg gagctgaacg gcttctgggc caacggcctg
     2101 aacagggagg gcgccatcag gatcggcgag gacgacggcg cccgaccggt agtagcagtg
     2161 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     2221 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     2281 ctgaccctga agttcatttg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     2341 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     2401 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     2461 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     2521 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     2581 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     2641 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     2701 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     2761 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     2821 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagcggcc
     2881 gcaattccaa ctgagcgccg gtcgctacca ttaccaactt gtctggtgtc aaaaataata
     2941 ggggccgctg tcatcagagt aagtttaaac tgagttctac taactaacga gtaatattta
     3001 aattttcagc atctcgcgcc cgtgcctctg acttctaagt ccaattactc ttcaacatcc
     3061 ctacatgctc tttctccctg tgctcccacc ccctattttt gttattatca aaaaacttct
     3121 cttaatttct ttgtttttta gcttctttta agtcacctct aacaatgaaa ttgtgtagat
     3181 tcaaaaatag aattaattcg taataaaaag tcgaaaaaaa ttgtgctccc tccccccatt
     3241 aataataatt ctatcccaaa atctacacaa tgttctgtgt acacttctta tgtttttttt
     3301 acttctgata aatttttttt gaaacatcat agaaaaaacc gcacacaaaa taccttatca
     3361 tatgttacgt ttcagtttat gaccgcaatt tttatttctt cgcacgtctg ggcctctcat
     3421 gacgtcaaat catgctcatc gtgaaaaagt tttggagtat ttttggaatt tttcaatcaa
     3481 gtgaaagttt atgaaattaa ttttcctgct tttgcttttt gggggtttcc cctattgttt
     3541 gtcaagagtt tcgaggacgg cgtttttctt gctaaaatca caagtattga tgagcacgat
     3601 gcaagaaaga tcggaagaag gtttgggttt gaggctcagt ggaaggtgag tagaagttga
     3661 taatttgaaa gtggagtagt gtctatgggg tttttgcctt aaatgacaga atacattccc
     3721 aatataccaa acataactgt ttcctactag tcggccgtac gggccctttc gtctcgcgcg
     3781 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     3841 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     3901 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     3961 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc ggccttaagg
     4021 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     4081 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4141 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4201 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4261 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4321 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4381 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4441 cggtattatc ccgtattgac gccggtgaag agcaactcgg tcgccgcata cactattctc
     4501 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4561 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4621 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4681 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4741 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4801 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4861 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4921 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4981 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5041 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5101 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5161 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5221 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5281 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5341 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     5401 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5461 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5521 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5581 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagcattgag
     5641 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5701 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5761 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5821 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5881 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     5941 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     6001 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     6061 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     6121 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     6181 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     6241 acgccaagct gtaagtttaa acatgatctt actaactaac tattctcatt taaattttca
     6301 gagcttaaaa atggctgaaa tcactcacaa cgatggatac gctaacaact tggaaatgaa
     6361 ataagct
//
