LOCUS       sequence-391257-        6557 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian cell expression of SNAPf-labeled TFAM.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6557)
  AUTHORS   Harding O, Holzer E, Riley JF, Martens S, Holzbaur ELF
  TITLE     Damaged mitochondria recruit the effector NEMO to activate NF-kappaB
            signaling.
  JOURNAL   Mol Cell. 2023 Sep 7;83(17):3188-3204.e7. doi: 
            10.1016/j.molcel.2023.08.005.
  PUBMED    37683611
REFERENCE   2  (bases 1 to 6557)
  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"; doi: 
            "10.1016/j.molcel.2023.08"; journalName: "Mol Cell"; date: 
            "2023-09-7- 7"; volume: "83"; issue: "17"; pages: "3188-3204.e7"
COMMENT     Sequence Label: TFAM-SNAPf
FEATURES             Location/Qualifiers
     source          1..6557
                     /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             1677..2222
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     intron          2333..2562
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     2545..2564
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(2635..3209,3210..3212,3213..3221)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2802..2819)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3029..3048
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3244..4047
                     /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(3307..3326)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3917..3936
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4177..4298
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4214..4233)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4400..4416)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4400..4416)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4413..4435)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4424..4440
                     /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(4448..4454,4455..4472,4473..4478))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4493..4514
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4631..4648)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4802..5390)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4882..4901)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5561..6352,6353..6421))
                     /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     6184..6203
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6422..6526)
                     /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 cgtcgctagc accatggcgt ttctccgaag catgtggggc gtgctgagtg
      961 ccctgggaag gtctggagca gagctgtgca ccggctgtgg aagtcgactg cgctccccct
     1021 tcagttttgt gtatttaccg aggtggtttt catctgtctt ggcaagttgt ccaaagaaac
     1081 ctgtaagttc ttaccttcga ttttctaaag aacaactacc catatttaaa gctcagaacc
     1141 cagatgcaaa aactacagaa ctaattagaa gaattgccca acgttggagg gaacttcctg
     1201 attcaaagaa aaaaatatat caagatgctt atagggcgga gtggcaggta tataaagaag
     1261 agatatgcag atttaaagaa cagctaactc caagtcagat tatgtctttg gaaaaagaaa
     1321 tcatggacaa acatttaaaa aggaaagcta tgacaaaaaa aaaagagtta acactgcttg
     1381 gaaaaccaaa aagacctcgt tcagcttata acgtttatgt agctgaaaga ttccaagaag
     1441 ctaagggtga ttcaccgcag gaaaagctga agactgtaaa ggaaaactgg aaaaatctgt
     1501 ctgactctga aaaggaatta tatattcagc atgctaaaga ggacgaaact cgttatcata
     1561 atgaaatgaa gtcttgggaa gaacaaatga ttgaagttgg acgaaaggat cttctacgtc
     1621 gcacaataaa gaaacaacga aaatatggtg ctgaggagtg taccggtgaa ttcaccatgg
     1681 acaaagactg cgaaatgaag cgcaccaccc tggatagccc tctgggcaag ctggaactgt
     1741 ctgggtgcga acagggcctg caccgtatca tcttcctggg caaaggaaca tctgccgccg
     1801 acgccgtgga agtgcctgcc ccagccgccg tgctgggcgg accagagcca ctgatgcagg
     1861 ccaccgcctg gctcaacgcc tactttcacc agcctgaggc catcgaggag ttccctgtgc
     1921 cagccctgca ccacccagtg ttccagcagg agagctttac ccgccaggtg ctgtggaaac
     1981 tgctgaaagt ggtgaagttc ggagaggtca tcagctacag ccacctggcc gccctggccg
     2041 gcaatcccgc cgccaccgcc gccgtgaaaa ccgccctgag cggaaatccc gtgcccattc
     2101 tgatcccctg ccaccgggtg gtgcagggcg acctggacgt ggggggctac gagggcgggc
     2161 tcgccgtgaa agagtggctg ctggcccacg agggccacag actgggcaag cctgggctgg
     2221 gtcctgcagg cggatccgcg tttaaactcg aggttaatta atgagcggcc gcatagataa
     2281 ctgatccagt gtgctggaat taattcgctg tctgcgaggg ccagctgttg gggtgagtac
     2341 tccctctcaa aagcgggcat gacttctgcg ctaagattgt cagtttccaa aaacgaggag
     2401 gatttgatat tcacctggcc cgcggtgatg cctttgaggg tggccgcgtc catctggtca
     2461 gaaaagacaa tctttttgtt gtcaagcttg aggtgtggca ggcttgagat ctggccatac
     2521 acttgagtga caatgacatc cactttgcct ttctctccac aggtgtccac tcccaggtcc
     2581 aactgcaggt cgagcatgca tctagggcgg ccaattccgc ccctctcccc cccccccctc
     2641 tccctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt
     2701 tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc
     2761 tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca
     2821 aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac
     2881 gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg
     2941 ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt
     3001 gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct
     3061 gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg
     3121 ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt
     3181 ggttttcctt tgaaaaacac gatgataagc ttgccacaac ccgggataat tcctgcagcc
     3241 aatatgggat cggccattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3301 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3361 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     3421 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     3481 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     3541 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     3601 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     3661 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     3721 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     3781 cgcatgcccg acggcgatga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     3841 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     3901 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     3961 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4021 tatcgccttc ttgacgagtt cttctgaggg gatcaattct ctagataact gatcataatc
     4081 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     4141 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     4201 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4261 tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtcgagtg cattctagtt
     4321 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     4381 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     4441 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     4501 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     4561 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     4621 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4681 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4741 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4801 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4861 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4921 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4981 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5041 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5101 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5161 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5221 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     5281 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5341 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5401 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5461 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5521 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5581 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5641 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5701 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5761 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5821 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5881 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5941 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6001 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6061 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6121 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6181 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6241 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6301 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6361 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6421 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6481 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6541 aagtgccacc tgacgtc
//
