LOCUS       sequence-208583-        6138 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  labeling of mitochondrial cytochrome c oxidase, pH measurements.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6138)
  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 6138)
  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: Cox7c-pHLuorin-sems
FEATURES             Location/Qualifiers
     source          1..6138
                     /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             1116..1829
                     /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(1146..1174)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1769..1789
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     intron          1920..2149
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     2132..2151
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    2327..2789
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2383..2400)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2610..2629
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2825..3628
                     /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(2888..2907)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3498..3517
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3758..3879
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3795..3814)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3981..3997)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3981..3997)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3994..4016)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4005..4021
                     /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(4029..4035,4036..4053,4054..4059))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4074..4095
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4212..4229)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4383..4971)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4463..4482)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5142..5933,5934..6002))
                     /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     5765..5784
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6003..6107)
                     /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 ttgggccaga gcatccggag gttcacaacc tctgtggtcc
      961 gtaggagcca ctatgaggag ggccctggga agaatttgcc attttcagtg gaaaacaagt
     1021 ggtcgttact agctaagatg tgtttgtact ttggatctgc atttgctaca cccttccttg
     1081 tagtaagaca ccaactgctt aaaacagaat tcaccatgag taaaggagaa gaacttttca
     1141 ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg
     1201 tcagtggaga gggtgaaggt gatgcaacat acggaaaact tacccttaaa tttatttgca
     1261 ctactggaaa actacctgtt ccttggccaa cacttgtcac tactttaact tatggtgttc
     1321 aatgcttttc aagataccca gatcatatga aacggcatga ctttttcaag agtgccatgc
     1381 ccgaaggtta tgtacaggaa agaactatat ttttcaaaga tgacgggaac tacaagacac
     1441 gtgctgaagt caagtttgaa ggtgataccc ttgttaatag aatcgagtta aaaggtattg
     1501 attttaaaga agatggaaac attcttggac acaaattgga atacaactat aacgatcacc
     1561 aggtgtacat catggcagac aaacaaaaga atggaatcaa agctaacttc aaaattagac
     1621 acaacattga agatggaggc gttcaactag cagaccatta tcaacaaaat actccaattg
     1681 gcgatgggcc cgtcctttta ccagacaacc attacctgtt tacaacttct actctttcga
     1741 aagatcccaa cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga
     1801 ttacacatgg catggatgaa ctatacaaaa ccggtggatc cctcgagtga ggcggccgca
     1861 tagataactg atccagtgtg ctggaattaa ttcgctgtct gcgagggcca gctgttgggg
     1921 tgagtactcc ctctcaaaag cgggcatgac ttctgcgcta agattgtcag tttccaaaaa
     1981 cgaggaggat ttgatattca cctggcccgc ggtgatgcct ttgagggtgg ccgcgtccat
     2041 ctggtcagaa aagacaatct ttttgttgtc aagcttgagg tgtggcaggc ttgagatctg
     2101 gccatacact tgagtgacaa tgacatccac tttgcctttc tctccacagg tgtccactcc
     2161 caggtccaac tgcaggtcga gcatgcatct agggcggcca attccgcccc tctccccccc
     2221 ccccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
     2281 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     2341 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     2401 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
     2461 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     2521 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     2581 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
     2641 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat
     2701 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
     2761 tggttttcct ttgaaaaaca cgatgataag cttgccacaa cccgggataa ttcctgcagc
     2821 caatatggga tcggccattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     2881 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     2941 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     3001 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     3061 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     3121 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     3181 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     3241 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     3301 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     3361 gcgcatgccc gacggcgatg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     3421 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     3481 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     3541 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     3601 ctatcgcctt cttgacgagt tcttctgagg ggatcaattc tctagataac tgatcataat
     3661 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     3721 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     3781 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     3841 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtcgagt gcattctagt
     3901 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     3961 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     4021 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     4081 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4141 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4201 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4261 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4321 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4381 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4441 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4501 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4561 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4621 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4681 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4741 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4801 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4861 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4921 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4981 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5041 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5101 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5161 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5221 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5281 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5341 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5401 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5461 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5521 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5581 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5641 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5701 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5761 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5821 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5881 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5941 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6001 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6061 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6121 aaagtgccac ctgacgtc
//
