LOCUS       sequence-452476-        4673 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of His-tagged SLC25A46 in Pichia pastoris.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4673)
  AUTHORS   Boopathy S, Luce BE, Lugo CM, Hakim P, McDonald J, Kim HL, Ponce J, 
            Ueberheide BM, Chao LH
  TITLE     Identification of SLC25A46 interaction interfaces with mitochondrial
            membrane fusogens Opa1 and Mfn2.
  JOURNAL   J Biol Chem. 2024 Aug 31:107740. doi: 10.1016/j.jbc.2024.107740.
  PUBMED    39222684
REFERENCE   2  (bases 1 to 4673)
  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: "J Biol 
            Chem. 2024 Aug 31:107740. doi: 10.1016/j.jbc.2024.107740."
COMMENT     Sequence Label: pPICZ A-SLC25A46-His (SB250)
FEATURES             Location/Qualifiers
     source          1..4673
                     /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"
     terminator      complement(664..911)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     877..895
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(977..1351)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(1370..1417)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(1424..1837)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      complement(1852..2098)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             complement(2177..2194)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(2210..2239)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(2263..2292)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             complement(2293..2313)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     promoter        complement(3654..4592)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
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 tggtctccag
      661 cttgcaaatt aaagccttcg agcgtcccaa aaccttctca agcaaggttt tcagtataat
      721 gttacatgcg tacacgcgtc tgtacagaaa aaaaagaaaa atttgaaata taaataacgt
      781 tcttaatact aacataacta taaaaaaata aatagggacc tagacttcag gttgtctaac
      841 tccttccttt tcggttagag cggatgtggg gggagggcgt gaatgtaagc gtgacataac
      901 taattacatg atatcgacaa aggaaaaggg ggacggatct ccgaggcctc ggacccgtcg
      961 ggccgcgtcg gacgtgtcag tcctgctcct cggccacgaa gtgcacgcag ttgccggccg
     1021 ggtcgcgcag ggcgaactcc cgcccccacg gctgctcgcc gatctcggtc atggccggcc
     1081 cggaggcgtc ccggaagttc gtggacacga cctccgacca ctcggcgtac agctcgtcca
     1141 ggccgcgcac ccacacccag gccagggtgt tgtccggcac cacctggtcc tggaccgcgc
     1201 tgatgaacag ggtcacgtcg tcccggacca caccggcgaa gtcgtcctcc acgaagtccc
     1261 gggagaaccc gagccggtcg gtccagaact cgaccgctcc ggcgacgtcg cgcgcggtga
     1321 gcaccggaac ggcactggtc aacttggcca tggtttagtt cctcaccttg tcgtattata
     1381 ctatgccgat atactatgcc gatgattaat tgtcaacacc gcccttagat tagattgcta
     1441 tgctttcttt ctaatgagca agaagtaaaa aaagttgtaa tagaacaaga aaaatgaaac
     1501 tgaaacttga gaaattgaag accgtttatt aacttaaata tcaatgggag gtcatcgaaa
     1561 gagaaaaaaa tcaaaaaaaa aaattttcaa gaaaaagaaa cgtgataaaa atttttattg
     1621 cctttttcga cgaagaaaaa gaaacgaggc ggtctctttt ttcttttcca aacctttagt
     1681 acgggtaatt aacgacaccc tagaggaaga aagaggggaa atttagtatg ctgtgcttgg
     1741 gtgttttgaa gtggtacggc gatgcgcgga gtccgagaaa atctggaaga gtaaaaaagg
     1801 agtagaaaca ttttgaagct atggtgtgtg ggggatccgc acaaacgaag gtctcactta
     1861 atcttctgta ctctgaagag gagtgggaaa taccaagaaa aacatcaaac tcgaatgatt
     1921 ttcccaaacc cctaccacaa gatattcatc agctgcgaga taggctgatc aggagcaagc
     1981 tcgtacgaga agaaacaaaa tgacaaaaaa aatcctatac tatataggtt acaaataaaa
     2041 aagtatcaaa aatgaagcct gcatctctca ggcaaatggc attctgacat cctcttgatt
     2101 agaatctagc aagaccggtc ttctcgtaag tgcccaactt gaactgagga acagtcatgt
     2161 ctaaggctaa aactcaatga tgatgatgat gatggtcgac ggcgctattc agatcctctt
     2221 ctgagatgag tttttgttcg ggcccaagct ggcggccgcc taatggtggt ggtgatgatg
     2281 gtgatgatga tgtccctgga aataaagatt ctcggaacca ccaccaccag aaccaccacc
     2341 accagaacca ccaccaccga tattgttttg caacaaagta gagtagatga tcttagtgat
     2401 ttgcaaaaca gcagcatgca aagtgtattg gataataaca gcaccgaaac ccttgtagaa
     2461 accaaaaaca ccctcttctt gtctgatagt gttgatacaa tctctcatac cctcatattg
     2521 agtattaata ggcaaaactt cgtaacccaa atcagtgtta tcgatgatag ttctagtacc
     2581 ttggatgtgc aatctatgca aaacagtttc caatgggtac aagataacat cagaacacaa
     2641 agaagcagcg aaattagcaa tcaactcagg aaaataagca tccaacatag attgaactgg
     2701 agaagtagat tcagccaagt gagagttgta agtctttctc ttcaagatca acaaaacgaa
     2761 cttttgaata acagaagaga tgatgtagtg caaaacacca tgcaaaacag tagggaagat
     2821 caaagacaac aatggcaaca atcttttaga atgaggaaca cccataccaa taactctacc
     2881 aataccctcc ttaacacatt ccaagatacc agtgttatct ctgatgatct cagattgaac
     2941 agtttcgatc aaagaagcag agtagaaagg catagcaaca acgtaagtca aagacttcaa
     3001 caacaagtgc tctccaattt gttttggaga ccacttatgc aaaacttctc taggcaatgg
     3061 agtgaactca gaaataatac cttcagcacc caaagtaaca ccttgaacaa taaaagtaga
     3121 acccataccc ttccacaaag ctctaggacc ttgagtctta ttgaaagagt acatgatgtt
     3181 aataacagtg aatggagtca agtgataatg ttgagcgtgg taattaactt gacattgtct
     3241 tctcaaaacg atacatggat gagccaaaac gttctcagtg aacaaagaag ccaaaccaat
     3301 accgaaacca gcaaatctat tcaattgttc agaagattga ccttgaacag aaccaccacc
     3361 accagaagaa aatggctctt cagtaggacc ctcatatgga gtagaagtag taggaacacc
     3421 gtaaggtgga gacttttcac cccagtgcaa gtttctagaa cctggaatat caggtggagt
     3481 agtaacccaa tgacccaaat cagaaccagt agaaaaagat ctagcaggga aagctcctcc
     3541 aaatccttgc tcgtctctgg ctcctcctct gtatcccaaa ccgtcaaatc cgtcaggtct
     3601 tctagggtgc atggtaccga tccgagacgg ccggctgggc cacgtgaatt cctcgtttcg
     3661 aataattagt tgttttttga tcttctcaag ttgtcgttaa aagtcgttaa aatcaaaagc
     3721 ttgtcaattg gaaccagtcg caattatgaa agtaagctaa taatgatgat aaaaaaaaag
     3781 gtttaagaca gggcagcttc cttctgttta tatattgctg tcaagtaggg gttagaacag
     3841 ttaaattttg atcatgaacg ttaggctatc agcagtattc ccaccagaat cttggaagca
     3901 tacaatgtgg agacaatgca taatcatcca aaaagcgggt gtttccccat ttgcgtttcg
     3961 gcacaggtgc accggggttc agaagcgata gagagactgc gctaagcatt aatgagatta
     4021 tttttgagca ttcgtcaatc aataccaaac aagacaaacg gtatgccgac ttttggaagt
     4081 ttctttttga ccaactggcc gttagcattt caacgaacca aacttagttc atcttggatg
     4141 agatcacgct tttgtcatat taggttccaa gacagcgttt aaactgtcag ttttgggcca
     4201 tttggggaac atgaaactat ttgaccccac actcagaaag ccctcatctg gagtgatgtt
     4261 cgggtgtaat gcggagcttg ttgcattcgg aaataaacaa acatgaacct cgccaggggg
     4321 gccaggatag acaggctaat aaagtcatgg tgttagtagc ctaatagaag gaattggaat
     4381 gagcgagctc caatcaagcc caataactgg gctggttttt cgatggcaaa agtgggtgtt
     4441 gaggagaaga ggagtggagg tcctgcgttt gcaacggtct gctgctagtg tatcccctcc
     4501 tgttgcgttt ggcacttatg tgtgagaatg gacctgtgga tgtcggatgg caaaaaggtt
     4561 tcattcaacc tttcgtcttt ggatgttaga tctgatctca tgaccaaaat cccttaacgt
     4621 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttc
//
