LOCUS       sequence-475759-        8150 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human ATP5F1B tagged with turboGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8150)
  AUTHORS   Rose K, Herrmann E, Kakudji E, Lizarrondo J, Celebi AY, Wilfling F, 
            Lewis SC, Hurley JH
  TITLE     In situ cryo-ET visualization of mitochondrial depolarization and 
            mitophagic engulfment.
  JOURNAL   Proc Natl Acad Sci U S A. 2025 Aug 5;122(31):e2511890122. doi: 
            10.1073/pnas.2511890122. Epub 2025 Jul 31.
  PUBMED    40743392
REFERENCE   2  (bases 1 to 8150)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2025-08-5"; volume: "122(31):e2511890122. 
            doi"; pages: " 10.1073/pnas.2511890122. Epub 2025 Jul 31"
COMMENT     Sequence Label: ATP5F1B-turboGFP
FEATURES             Location/Qualifiers
     source          1..8150
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1040..1161
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1051..1070)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1105..1124
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1335..2129)
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1446..1465)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2056..2075
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2196..2525)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2210..2345
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2510..2530
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    2554..3176
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     2958..2977
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     3187..3203
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3226..3921)
                     /codon_start=1
                     /product="improved green fluorescent protein from 
                     Pontellina plumata (Evdokimov et al., 2006)"
                     /label=TurboGFP
                     /note="mammalian codon-optimized"
                     /translation="MESDESGLPAMEIECRITGTLNGVEFELVGGGEGTPEQGRMTNKM
                     KSTKGALTFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVL
                     HVSFSYRYEAGRVIGDFKVMGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNDLDGSFTR
                     TFSLRDGGYYSSVVDSHMHFKSAIHPSILQNGGPMFAFRRVEEDHSNTELGIVEYQHAF
                     KTPDADAGEE"
     primer_bind     complement(5587..5606)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5588..5606)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(5632..5835)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5661..5681)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5836..6215
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(6376..6393)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6376..6392)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6385..6407)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(6533..6988)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6670..6691)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6882..6901
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        7015..7119
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7120..7188,7189..7980)
                     /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     complement(7338..7357)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt
     1021 cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     1081 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     1141 ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat ccccgcgctg
     1201 gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt catagaaggc
     1261 ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg tcatttcgaa
     1321 ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa
     1381 tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc gccaagctct
     1441 tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtcggccac acccagccgg
     1501 ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg caagcaggca
     1561 tcgccatggg tcacgacgag atcctcgccg tcgggcatgc gcgccttgag cctggcgaac
     1621 agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc gacaagaccg
     1681 gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag
     1741 gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga tactttctcg
     1801 gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa tagcagccag
     1861 tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc
     1921 agccacgata gccgcgctgc ctcgtcctgc agttcattca gggcaccgga caggtcggtc
     1981 ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc atcagagcag
     2041 ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc ggccggagaa
     2101 cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatca
     2161 gatccgaaaa tggatataca agctcccggg agctttttgc aaaagcctag gcctccaaaa
     2221 aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc ctctgcataa
     2281 ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc
     2341 gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc
     2401 atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga
     2461 tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga
     2521 cacacattcc acagccaagc tggccgcgta cccaattcaa caggcatcta ctgagtggac
     2581 ccaacgcatg agaggacagt gccaagcaag caactcaaat gtcccaccgg ttgggcatgg
     2641 ccaggtagcc tatgctgtgt ctggacgtcc tcctgctggt atagttattt taaaatcaga
     2701 aggacaggga agggagcagt ggttcacgcc tgtaatccca gcaatttggg aggccaaggt
     2761 gggtagatca cctgagatta ggagttggag accagcctgg ccaatatggt gaaaccccgt
     2821 ctctaccaaa aaaacaaaaa ttagctgagc ctggtcatgc atgcctggaa tcccaacaac
     2881 tcgggaggct gaggcaggag aatcgcttga acccaggagg cggagattgc agtgagccaa
     2941 gattgtgcca ctgcactcca gcttggttcc caatagaccc cgcaggccct acaggttgtc
     3001 ttcccaactt gccccttgct ccataccacc cccctccacc ccataatatt atagaaggac
     3061 acctagtcag acaaaatgat gcaacttaat tttattagga caaggctggt gggcactgga
     3121 gtggcaactt ccagggccag gagaggcact ggggaggggt cacagggatg ccacccggga
     3181 tctgttcagg aaacagctat gaccgcggcc ggccgtttaa actctttctt caccggcatc
     3241 tgcatccggg gtcttgaagg cgtgctggta ctccacgatg cccagctcgg tgttgctgtg
     3301 atcctcctcc acgcggcgga aggcgaacat ggggcccccg ttctgcagga tgctggggtg
     3361 gatggcgctc ttgaagtgca tgtggctgtc caccacggag ctgtagtagc cgccgtcgcg
     3421 caggctgaag gtgcgggtga agctgccatc cagatcgtta tcgcccatgg ggtgcaggtg
     3481 ctccacggtg gcgttgctgc ggatgatctt gtcggtgaag atcacgctgt cctcggggaa
     3541 gccggtgccc atcaccttga agtcgccgat cacgcggccg gcctcgtagc ggtagctgaa
     3601 gctcacgtgc agcacgccgc cgtcctcgta cttctcgatg cgggtgttgg tgtagccgcc
     3661 gttgttgatg gcgtgcagga aggggttctc gtagccgctg gggtaggtgc cgaagtggta
     3721 gaagccgtag cccatcacgt ggctcagcag gtaggggctg aaggtcaggg cgcctttggt
     3781 gctcttcatc ttgttggtca tgcggccctg ctcgggggtg ccctctccgc cgcccaccag
     3841 ctcgaactcc acgccgttca gggtgccggt gatgcggcac tcgatctcca tggcgggcag
     3901 gccgctctcg tcgctctcca tctcgagcgg ccgcgtacgc gtcgatgaat gctcttcagc
     3961 cagcttatca gcttttgcca cagcttcttc aatgggtccc accatataga aggcctgttc
     4021 tgggagatgg tcatattcac ctgccaaaat ctgctggaat cctttgatgg tctccttcag
     4081 gggtaccagc ttccccatat gacctgtgaa gacctcagca acctggaatg gctgagacaa
     4141 gaaacgctgt attttccgtg cacgggacac ggtcaacttg tcttcctcag aaagttcatc
     4201 catacccagg atggcaatga tatcctggag ggatttgtag tcctgcagga tcttttgcac
     4261 cccacgggca acatcgtaat gctcactgcc aacaatgttg ggatccatga tacgagaggt
     4321 ggagtctaga ggatccacag ctggatagat gcccagctca gcaatggcac gcgacagtac
     4381 agtggtagca tccaaatggg caaacgtagt agcaggggca gggtcagtca agtcatcagc
     4441 aggcacatag atagcctgta cagaggtgat agatcccttc ttggtagtgg taattctttc
     4501 ctgcatagta cccatgtcag tggccagggt aggctgatag cccacagcag aagggattcg
     4561 gcccaataat gcagacacct ctgaaccagc ctgggtgaag cgaaagatgt tatcaataaa
     4621 tagcagtaca tcttgacctt cttggtctct gaagtattca gccacagtca gcccagtcag
     4681 agctacccgg gcacgagcac caggtggttc attcatttga ccatatacca gcgctacctt
     4741 agaggtggca tcttttaagt tgataacacc agattcaatc atttcatggt ataaatcatt
     4801 gccttcacgg gtcctctcac caacaccagc aaacacagag taaccaccat gggctttggc
     4861 gacattgttg attaactcca tgatcagtac agtcttgcca actccagcac caccaaaaag
     4921 cccaattttg ccacccttgg catagggagc tagcagatcg acaaccttga taccagtcac
     4981 cagaatttcc tgctcaacac tcatttccat gaactctgga gcctcagcat gaatgggagc
     5041 aaattgtttg gttttgatgg gacctctttc atcaataggt tctccaatga cattcatgat
     5101 tctgcccaaa gtctcaggac caacaggaat tttgattggt gcaccagaat ccagtacttt
     5161 ctggcctcta accaagcctt ctgtaccatc catagcaata gtccttactg tgctctcacc
     5221 caaatgctgg gccacctcca aaaccagtct ggtctccctg ccttgcactt ccagggcatt
     5281 tagaattggt ggtagtccct catcaaactg gacgtccacc actgcgccaa tgaccgccac
     5341 gatgcgcccg gtggcggcgc ctgcttttgg cgaaggagat gtttgcgccg catagtccct
     5401 gacaggatgg accgccgtcg gagcggcccg cagtaagagc tgagctgggg gcagcgacgc
     5461 tgaaggggtg agtctccgca aggccccgga ggccggagca gcggccaccc gacccacaaa
     5521 ccccaacatg gcgatcgcgg cggcagatct cctcggtacc ggatccagtc gacgaattcc
     5581 cggccgccct atagtgagtc gtattacaaa attctgacgg ttcactaaac gagctctgct
     5641 tatatagacc tcccaccgta cacgcctacc gcccatttgc gtcaacgggg cggggttatt
     5701 acgacatttt ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat
     5761 ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tggtgtactg
     5821 ccaaaaccgc atcaccatgg taatagcgat gactaatacg tagatgtact gccaagtagg
     5881 aaagtcccgt aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac
     5941 gtcaataggg ggcggacttg gcatatgata cacttgatgt actgccaagt gggcagttta
     6001 ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac
     6061 atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt
     6121 taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat
     6181 tgattactat taataactag tcaataatca atgtcaacat ggcggtcata ttggacatga
     6241 gccaatataa atgtacatat tatgatatag atacaacgta tgcaatggcc aatagccaat
     6301 attgatttat gctatataac caatgactaa tatggctaat tgccaatatt gattcaatgt
     6361 atagatcagc ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     6421 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     6481 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggaaattgta
     6541 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     6601 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     6661 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     6721 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     6781 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     6841 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     6901 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     6961 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     7021 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     7081 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     7141 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     7201 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     7261 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     7321 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     7381 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     7441 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     7501 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     7561 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     7621 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     7681 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     7741 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     7801 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     7861 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     7921 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     7981 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     8041 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     8101 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
//
