LOCUS       sequence-485508-        5889 bp    DNA     circular SYN 08-SEP-2025
DEFINITION  endogenously tags TOMM70 with Halo tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5889)
  AUTHORS   Thayer JA, Petersen JD, Huang X, Hawrot J, Ramos DM, Sekine S, Li Y,
            Ward ME, Narendra DP
  TITLE     Novel reporter of the PINK1-Parkin mitophagy pathway identifies its 
            damage sensor in the import gate.
  JOURNAL   bioRxiv [Preprint]. 2025 Feb 20:2025.02.19.639160. doi: 
            10.1101/2025.02.19.639160.
  PUBMED    40027798
REFERENCE   2  (bases 1 to 5889)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 8, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2025 Feb 20:2025.02.19.639160. doi: 
            10.1101/2025.02.19.639160."
COMMENT     Sequence Label: TOMM70-FKBP12-Halo donor plasmid
FEATURES             Location/Qualifiers
     source          1..5889
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             588..908
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             927..1814
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="AEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
                     EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
                     GSEIARWLSTLEISG"
     primer_bind     complement(2836..2852)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2836..2852)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2849..2871)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2860..2876
                     /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(2884..2890,2891..2908,2909..2914))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2929..2950
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3067..3084)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3238..3826)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3318..3337)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3997..4788,4789..4857))
                     /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     4620..4639
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4858..4962)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5030..5048
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5086..5108)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5208..5227
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5421..5443
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5435..5452
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5436..5452
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cggtattatg tcactgcttt ctaaaggagt ctcatgttaa cagtagaaac atgtaaaatt
       61 taggaaccag acggaattgt gattttaaat aagccaatca atttccctag tctgactgtt
      121 cttatcagta aacacagaat acaagggata taatgtaagc acatagtatt tgtaatagta
      181 gttgacattc attgagtgct ttcaatgtgg taggtactct tgtttatttt aaaagaagca
      241 ttttactaga atcatagtag aaacttagtg gtcagacttc atgtagcatc acttacaacc
      301 ttcattgaat attgtgaaga aacttagtgg tcagacttca tgtaccatca cttacaacct
      361 tcattgaata ttgtgaagtg cttgttgaat ctgagactta aaaattttgt tcttttagag
      421 gaaacatgga gaaagccatt gacatgttca acaaagctat taacctggcc aaatcggaaa
      481 tggagatggc ccatctgtat tcactttgcg atgccgccca tgcccagaca gaagttgcaa
      541 agaaatacgg attaaaaccg ccgacgctgg gaggatcggg aggatcagga gtgcaggtgg
      601 aaaccatctc cccaggagac gggcgcacct tccccaagcg cggccagacc tgcgtggtgc
      661 actacaccgg gatgcttgaa gatggaaaga aatttgattc ctcccgggac agaaacaagc
      721 cctttaagtt tatgctaggc aagcaggagg tgatccgagg ctgggaagaa ggggttgccc
      781 agatgagtgt gggtcagaga gccaaactga ctatatctcc agattatgcc tatggtgcca
      841 ctgggcaccc aggcatcatc ccaccacatg ccactctcgt cttcgatgtg gagcttctaa
      901 aactggaagg aggatcggga ggatcagcag aaatcggtac tggctttcca ttcgaccccc
      961 attatgtgga agtcctgggc gagcgcatgc actacgtcga tgttggtccg cgcgatggca
     1021 cccctgtgct gttcctgcac ggtaacccga cctcctccta cgtgtggcgc aacatcatcc
     1081 cgcatgttgc accgacccat cgctgcattg ctccagacct gatcggtatg ggcaaatccg
     1141 acaaaccaga cctgggttat ttcttcgacg accacgtccg cttcatggat gccttcatcg
     1201 aagccctggg tctggaagag gtcgtcctgg tcattcacga ctggggctcc gctctgggtt
     1261 tccactgggc caagcgcaat ccagagcgcg tcaaaggtat tgcatttatg gagttcatcc
     1321 gccctatccc gacctgggac gaatggccag aatttgcccg cgagaccttc caggccttcc
     1381 gcaccaccga cgtcggccgc aagctgatca tcgatcagaa cgtttttatc gagggtacgc
     1441 tgccgatggg tgtcgtccgc ccgctgactg aagtcgagat ggaccattac cgcgagccgt
     1501 tcctgaatcc tgttgaccgc gagccactgt ggcgcttccc aaacgagctg ccaatcgccg
     1561 gtgagccagc gaacatcgtc gcgctggtcg aagaatacat ggactggctg caccagtccc
     1621 ctgtcccgaa gctgctgttc tggggcaccc caggcgttct gatcccaccg gccgaagccg
     1681 ctcgcctggc caaaagcctg cctaactgca aggctgtgga catcggcccg ggtctgaatc
     1741 tgctgcaaga agacaacccg gacctgatcg gcagcgagat cgcgcgctgg ctgtcgacgc
     1801 tcgagatttc cggctaaaat agagggaaag cagactgacc ctctttttaa aagtttaccc
     1861 cctcttcaac tgaaccctaa agacactgtc atgaactgtg ttgaatggtg gaaatcagta
     1921 tttctgtttg tggtgttgtt atttgttaca tctgtttcat gtctaggtgt tgtgggtgtg
     1981 gctgttgaag gaagtttgca gtcttgcagc ttttattccc tgtgcaacaa aagattagaa
     2041 catgttaaag ggatttttaa ataaagttgc aaagagtaca aatgataatt ggccatgcaa
     2101 ataaaaactg atttgttgat ttttttttta aggggggttg gcagttgatt atgttctgga
     2161 tgattccgtc tatatatgtg tgaataatgt aagtatttta cagcatgttg atttttaaat
     2221 taacgtagta aatgctgtaa aatagattta tattcagtta accgctttca gttgattttt
     2281 tgaaagaaac aaaggttaaa tgggggatta aagtaaaatt gagagaccct ttaaaccatt
     2341 gtcagcatgc acaatgcctc tgattctgca gttttagaaa cttggtggca cttattaatc
     2401 ctcttggccc ctttccactc taatggatag tgtacattct tcttaaagtc cacaacagca
     2461 gattttcttg cagtaaatta tgcagatgca aaatattcta attgatatat gtgttggaag
     2521 actgagtatt gatgggggag tggaccagac aaagaggtaa gatgaaacag tagtgtgttt
     2581 ataattgtct gtgactattt tctataataa ttagtactat ttaatggtga gcttttaaaa
     2641 atgtaggata gagggtacag tggcactgta tatactattt atagtctcag ctactaggga
     2701 ggctgaggca agaggcttga gaccaggagc tcgaggctgt aatgtgccat gatgattgca
     2761 cctgcgaata gccactgcac tccagcctgg gcaatatagc aagaccccat cctttaaaag
     2821 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     2881 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     2941 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3001 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3061 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3121 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3181 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3241 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3301 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3361 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3421 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3481 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3541 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3601 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3661 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3721 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3781 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3841 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3901 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3961 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4021 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4081 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4141 tccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4201 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4261 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4321 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4381 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4441 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4501 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4561 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4621 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4681 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4741 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4801 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4861 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4921 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4981 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     5041 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     5101 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     5161 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     5221 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     5281 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     5341 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     5401 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattcct
     5461 atcacgttgc catatattat aatgggggat taaatatagt caccctagtg gtacctctta
     5521 ctggcatata aagtctaact ggcacatatg ccaataaaat gtaccttact aacatgtatg
     5581 ccagttagac tctgtatatg caggtatata tatatatagt agggacttaa tattctttgt
     5641 aggattgcat atatatgtag ctaatactag ttacttaaaa cagttggttc ttagatagct
     5701 gtggatttga ccagaatcag gcagaggcaa taaatctgag tatttattga gtaggtgtga
     5761 ggaactcaca tagactgcct gaagagtaga gatgcatggg gttggggatg tgctggtaag
     5821 agatgaaaca gaaagaggtg ggcccacatc agtctgtgaa ggtaaggccc ttgcatgtca
     5881 ttagtgtcc
//
