LOCUS       sequence-208581-        6456 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  labeling of mitochondrial succinate dehydrogenase subunit C, pH 
            measurements.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6456)
  AUTHORS   Rieger B, Shalaeva DN, Sohnel AC, Kohl W, Duwe P, Mulkidjanian AY, 
            Busch KB
  TITLE     Lifetime imaging of GFP at CoxVIIIa reports respiratory supercomplex
            assembly in live cells.
  JOURNAL   Sci Rep. 2017 Apr 6;7:46055. doi: 10.1038/srep46055.
  PUBMED    28383048
REFERENCE   2  (bases 1 to 6456)
  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: "Sci Rep."; 
            date: "2017-04-6"; volume: "7:46055. doi"; pages: " 
            10.1038/srep46055"
COMMENT     Sequence Label: SDHC-pHLuorin-sems
FEATURES             Location/Qualifiers
     source          1..6456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1434..2147
                     /codon_start=1
                     /product="pH-sensitive mutant of green fluorescent protein 
                     (Ng et al., 2002)"
                     /label=superecliptic pHluorin
                     /note="excited strongly by blue light at neutral pH but 
                     minimally fluorescent at acidic pH"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNDHQVYIMADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLFTTSTLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1464..1492)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2087..2107
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     intron          2238..2467
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     2450..2469
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    2645..3107
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2701..2718)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2928..2947
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3143..3946
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3206..3225)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3816..3835
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4076..4197
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4113..4132)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4299..4315)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4299..4315)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4312..4334)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4323..4339
                     /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(4347..4353,4354..4371,4372..4377))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4392..4413
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4530..4547)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4701..5289)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4781..4800)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5460..6251,6252..6320))
                     /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     6083..6102
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6321..6425)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat cgatatcatg gctgcgctgt tgctgagaca cgttggtcgt cattgcctcc
      961 gagcccactt tagccctcag ctctgtatca gaaatgctgt tcctttggga accacggcca
     1021 aagaagagat ggagcggttc tggaataaga atataggttc aaaccgtcct ctgtctcccc
     1081 acattactat ctacagttgg tctcttccca tggcgatgtc catctgccac cgtggcactg
     1141 gtattgcttt gagtgcaggg gtctctcttt ttggcatgtc ggccctgtta ctccctggga
     1201 actttgagtc ttatttggaa cttgtgaagt ccctgtgtct ggggccagca ctgatccaca
     1261 cagctaagtt tgcacttgtc ttccctctca tgtatcatac ctggaatggg atccgacact
     1321 tgatgtggga cctaggaaaa ggcctgaaga ttccccagct ataccagtct ggagtggttg
     1381 tcctggttct tactgtgttg tcctctatgg ggctggcagc catggaattc accatgagta
     1441 aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat ggtgatgtta
     1501 atgggcacaa attttctgtc agtggagagg gtgaaggtga tgcaacatac ggaaaactta
     1561 cccttaaatt tatttgcact actggaaaac tacctgttcc ttggccaaca cttgtcacta
     1621 ctttaactta tggtgttcaa tgcttttcaa gatacccaga tcatatgaaa cggcatgact
     1681 ttttcaagag tgccatgccc gaaggttatg tacaggaaag aactatattt ttcaaagatg
     1741 acgggaacta caagacacgt gctgaagtca agtttgaagg tgataccctt gttaatagaa
     1801 tcgagttaaa aggtattgat tttaaagaag atggaaacat tcttggacac aaattggaat
     1861 acaactataa cgatcaccag gtgtacatca tggcagacaa acaaaagaat ggaatcaaag
     1921 ctaacttcaa aattagacac aacattgaag atggaggcgt tcaactagca gaccattatc
     1981 aacaaaatac tccaattggc gatgggcccg tccttttacc agacaaccat tacctgttta
     2041 caacttctac tctttcgaaa gatcccaacg aaaagagaga ccacatggtc cttcttgagt
     2101 ttgtaacagc tgctgggatt acacatggca tggatgaact atacaaaacc ggtggatccc
     2161 tcgagtgagg cggccgcata gataactgat ccagtgtgct ggaattaatt cgctgtctgc
     2221 gagggccagc tgttggggtg agtactccct ctcaaaagcg ggcatgactt ctgcgctaag
     2281 attgtcagtt tccaaaaacg aggaggattt gatattcacc tggcccgcgg tgatgccttt
     2341 gagggtggcc gcgtccatct ggtcagaaaa gacaatcttt ttgttgtcaa gcttgaggtg
     2401 tggcaggctt gagatctggc catacacttg agtgacaatg acatccactt tgcctttctc
     2461 tccacaggtg tccactccca ggtccaactg caggtcgagc atgcatctag ggcggccaat
     2521 tccgcccctc tccccccccc ccctcccccc cccctaacgt tactggccga agccgcttgg
     2581 aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca
     2641 atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc
     2701 ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag
     2761 cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg
     2821 gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac
     2881 aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa
     2941 gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc
     3001 tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc
     3061 ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataagct tgccacaacc
     3121 cgggataatt cctgcagcca atatgggatc ggccattgaa caagatggat tgcacgcagg
     3181 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3241 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     3301 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     3361 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     3421 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     3481 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     3541 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     3601 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     3661 gttcgccagg ctcaaggcgc gcatgcccga cggcgatgat ctcgtcgtga cccatggcga
     3721 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     3781 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     3841 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     3901 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagggg atcaattctc
     3961 tagataactg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     4021 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     4081 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4141 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg
     4201 cgtcgagtgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     4261 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     4321 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4381 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4441 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4501 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4561 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4621 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4681 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4741 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4801 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4861 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4921 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4981 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5041 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5101 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5161 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5221 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5281 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5341 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5401 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5461 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5521 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5581 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5641 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5701 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5761 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5821 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5881 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5941 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6001 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6061 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6121 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6181 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6241 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6301 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6361 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6421 ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
