LOCUS       sequence-417747-        8758 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentivirus backbone expressing human TFAM fused with BioID2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8758)
  AUTHORS   Fu Y, Sacco O, DeBitetto E, Kanshin E, Ueberheide B, Sfeir A
  TITLE     Mitochondrial DNA breaks activate an integrated stress response to 
            reestablish homeostasis.
  JOURNAL   Mol Cell. 2023 Oct 8:S1097-2765(23)00753-0. doi: 
            10.1016/j.molcel.2023.09.026.
  PUBMED    37832546
REFERENCE   2  (bases 1 to 8758)
  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: "Mol Cell. 
            2023 Oct 8:S1097-2765(23)00753-0. doi: 
            10.1016/j.molcel.2023.09.026."
COMMENT     Sequence Label: lenti-CMV-TFAM-BioID2-IRES-Puro
FEATURES             Location/Qualifiers
     source          1..8758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        2..381
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..585
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             1413..1460
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     CDS             1503..2198
                     /codon_start=1
                     /product="promiscuous R40G mutant of a biotin protein 
                     ligase from Aquifex aeolicus (Kim et al., 2016)"
                     /label=BioID2
                     /translation="FKNLIWLKEVDSTQERLKEWNVSYGTALVADRQTKGRGGLGRKWL
                     SQEGGLYFSFLLNPKEFENLLQLPLVLGLSVSEALEEITEIPFSLKWPNDVYFQEKKVS
                     GVLCELSKDKLIVGIGINVNQREIPEEIKDRATTLYEITGKDWDRKEVLLKVLKRISEN
                     LKKFKEKSFKEFKGKIESKMLYLGEEVKLLGEGKITGKLVGLSEKGGALILTEEGIKEI
                     LSGEFSLRRS"
     CDS             2199..2225
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    2465..2926
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2521..2538)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2748..2767
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2935..3534
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2935..2954)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3431..3451
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(3552..3569)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3558..3782
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     LTR             4146..4326
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4388..4976)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4468..4487)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5147..5938,5939..6007))
                     /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     5770..5789
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6008..6112)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6187..6206)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6378..6757
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6759..6957
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             6975..7155
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7202..7327
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7824..8057
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8242..8286
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    8584..8701
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 agacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
       61 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      121 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      181 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      241 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      301 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      361 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
      421 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      481 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      541 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgcgg gttccagttg
      601 tgattgctgg agttgtgtat tgccaggagg ctctccgaga ttggggtcgg gtcactgcct
      661 catccaccgg agcgatggcg tttctccgaa gcatgtgggg cgtgctgagt gccctgggaa
      721 ggtctggagc agagctgtgc accggctgtg gaagtcgact gcgctccccc ttcagttttg
      781 tgtatttacc gaggtggttt tcatctgtct tggcaagttg tccaaagaaa cctgtaagtt
      841 cttaccttcg attttctaaa gaacaactac ccatatttaa agctcagaac ccagatgcaa
      901 aaactacaga actaattaga agaattgccc agcgttggag ggaacttcct gattcaaaga
      961 aaaaaatata tcaagatgct tatagggcgg agtggcaggt atataaagaa gagataagca
     1021 gatttaaaga acagctaact ccaagtcaga ttatgtcttt ggaaaaagaa atcatggaca
     1081 aacatttaaa aaggaaagct atgacaaaaa aaaaagagtt aacactgctt ggaaaaccaa
     1141 aaagacctcg ttcagcttat aacgtttatg tagctgaaag attccaagaa gctaagggtg
     1201 attcaccgca ggaaaagctg aagactgtaa aggaaaactg gaaaaatctg tctgactctg
     1261 aaaaggaatt atatattcag catgctaaag aggacgaaac tcgttatcat aatgaaatga
     1321 agtcttggga agaacaaatg attgaagttg gacgaaagga tcttctacgt cgcacaataa
     1381 agaaacaacg aaaatatggt gctgaggagt gtgactacaa ggacgacgat gacaaggact
     1441 acaaggacga cgatgacaag gaattggcta gcggaggcgg tgggtcggga ggtggcggat
     1501 ccttcaagaa cctgatctgg ctgaaggagg tggacagcac ccaggagaga ctgaaggagt
     1561 ggaacgtgag ctacggcacc gccctggtgg ccgacagaca gaccaagggc agaggcggcc
     1621 tgggcagaaa gtggctgagc caggagggcg gcctgtactt cagcttcctg ctgaacccca
     1681 aggagttcga gaacctgctg cagctgcccc tggtgctggg cctgagcgtg agcgaggccc
     1741 tggaggagat caccgagatc cccttcagcc tgaagtggcc caacgacgtg tacttccagg
     1801 agaagaaggt gagcggcgtg ctgtgcgagc tgagcaagga caagctgatc gtgggcatcg
     1861 gcatcaacgt gaaccagaga gagatccccg aggagatcaa ggacagagcc accaccctgt
     1921 acgagatcac cggcaaggac tgggacagaa aggaggtgct gctgaaggtg ctgaagagaa
     1981 tcagcgagaa cctgaagaag ttcaaggaga agagcttcaa ggagttcaag ggcaagatcg
     2041 agagcaagat gctgtacctg ggcgaggagg tgaagctgct gggcgagggc aagatcaccg
     2101 gcaagctggt gggcctgagc gagaagggcg gcgccctgat cctgaccgag gagggcatca
     2161 aggagatcct gagcggcgag ttcagcctga gaagaagcta cccttacgat gtaccggatt
     2221 acgcatagaa gcttaagttt gtaaccacga tcgttatgct gatcataccc taatgatccc
     2281 agcaagataa tgtcctgtct tctaagatgt gcatcaagcc tggtacatac tgaaaaccct
     2341 ataaggtcct ggataatttt tgtttgatta ttcattgaag aaacatttat tttccaattg
     2401 tgtgaagttt ttgactgtta ataaaagaat ctgtcaacca tcaagaattc agggcccgtt
     2461 tgtagagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc
     2521 ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag
     2581 cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg
     2641 gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac
     2701 aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa
     2761 gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc
     2821 tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaac gtctaggccc
     2881 cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataagttt aaacatgacc
     2941 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     3001 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     3061 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     3121 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     3181 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     3241 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     3301 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     3361 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     3421 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     3481 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgatacaaa
     3541 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3601 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3661 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3721 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     3781 ggcagtctgt agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc
     3841 actcccaacg aagacaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc
     3901 ctgattggca gaactacaca ccagggccag ggatcagata tccactgacc tttggatggt
     3961 gctacaagct agtaccagtt gagcaagaga aggtagaaga agccaatgaa ggagagaaca
     4021 cccgcttgtt acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtattag
     4081 agtggaggtt tgacagccgc ctagcatttc atcacatggc ccgagagctg catccggact
     4141 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     4201 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     4261 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     4321 ctagcagcat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4381 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4441 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4501 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4561 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4621 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4681 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4741 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4801 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4861 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4921 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4981 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5041 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5101 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5161 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5221 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5281 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5341 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5401 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5461 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5521 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5581 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5641 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5701 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5761 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5821 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5881 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5941 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6001 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6061 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6121 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac
     6181 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     6241 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
     6301 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg
     6361 ggccagatat acgcgttgac attgattatt gactagttat taatagtaat caattacggg
     6421 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     6481 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     6541 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     6601 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     6661 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     6721 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
     6781 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     6841 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     6901 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagcgc
     6961 gttttgcctg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     7021 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     7081 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     7141 ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga
     7201 gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg
     7261 actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga
     7321 gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc
     7381 agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg
     7441 attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca
     7501 gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc
     7561 aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa
     7621 gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg gccgctgatc
     7681 ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa
     7741 gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag
     7801 agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga
     7861 agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt
     7921 atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa
     7981 ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta
     8041 aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct
     8101 gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc
     8161 tggatggagt gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa
     8221 gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca
     8281 agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg
     8341 atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga
     8401 gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc
     8461 gacaggcccg aaggaataga agaagaaggt ggagagagag acagagacag atccattcga
     8521 ttagtgaacg gatcggcact gcgtgcgcca attctgcaga caaatggcag tattcatcca
     8581 caattttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     8641 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     8701 tcgggtttat tacagggaca gcagagatcc agtttggtta gtaccgggcc cgctctag
//
