LOCUS       sequence-453893-        5378 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  plasmid encoding human SLC25A13 with an APEX protein tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5378)
  AUTHORS   Sha Z, Benkovic SJ
  TITLE     Purinosomes spatially co-localize with mitochondrial transporters.
  JOURNAL   J Biol Chem. 2024 Aug 2;300(9):107620. doi: 
            10.1016/j.jbc.2024.107620.
  PUBMED    39098527
REFERENCE   2  (bases 1 to 5378)
  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."; date: "2024-08-2"; volume: "300(9):107620. doi"; pages: " 
            10.1016/j.jbc.2024.107620"
COMMENT     Sequence Label: pSLC25A13-APEX2
FEATURES             Location/Qualifiers
     source          1..5378
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2708..3454
                     /codon_start=1
                     /product="soybean ascorbate peroxidase, engineered to serve
                     as a monomeric tag for electron microscopy and live-cell 
                     proteomics (Lam et al., 2015)"
                     /label=APEX2
                     /note="more active than the original APEX due to the A134P 
                     mutation"
                     /translation="GKSYPTVSADYQDAVEKAKKKLRGFIAEKRCAPLMLRLAFHSAGT
                     FDKGTKTGGPFGTIKHPAELAHSANNGLDIAVRLLEPLKAEFPILSYADFYQLAGVVAV
                     EVTGGPKVPFHPGREDKPEPPPEGRLPDPTKGSDHLRDVFGKAMGLTDQDIVALSGGHT
                     IGAAHKERSGFEGPWTSNPLIFDNSYFTELLSGEKEGLLQLPSDKALLSDPVFRPLVDK
                     YAADEDAFFADYAEAHQKLSELGFADA"
     CDS             3461..3502
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    complement(3503..3602)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3619..3638)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3620..3638)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3643..3659)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3643..3659)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3761..3780)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3772..4581
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3839..3858)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4728..5316
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5217..5236
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg ataagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc accatggcag cagcaaaggt ggccctgacc aagagggccg atcctgccga
      721 gctgcgcaca atcttcctga agtacgccag catcgagaag aacggcgagt tctttatgtc
      781 cccaaatgac tttgtgaccc ggtatctgaa catcttcggc gagtctcagc ccaatcctaa
      841 gaccgtggag ctgctgagcg gagtggtgga ccagacaaag gatggcctga tctccttcca
      901 ggagtttgtg gccttcgagt ctgtgctgtg cgccccagac gccctgttta tggtggcctt
      961 tcagctgttc gataaggccg gcaagggcga ggtgaccttc gaggacgtga agcaggtgtt
     1021 tggccagacc acaatccacc agcacatccc cttcaactgg gatagcgagt tcgtgcagct
     1081 gcactttggc aaggagcgga agagacacct gacctacgcc gagtttacac agttcctgct
     1141 ggagatccag ctggagcacg ccaagcaggc ctttgtgcag agagacaatg cccggaccgg
     1201 cagagtgaca gccatcgact ttcgggatat catggtgacc atcagacccc acgtgctgac
     1261 acctttcgtg gaggagtgtc tggtggcagc agcaggaggc accacatccc accaggtgtc
     1321 ttttagctac ttcaacggct ttaattctct gctgaacaat atggagctga tccggaagat
     1381 ctatagcacc ctggccggca caagaaagga cgtggaggtg accaaggagg agttcgtgct
     1441 ggccgcccag aagtttggcc aggtgacacc tatggaggtg gacatcctgt tccagctggc
     1501 cgatctgtac gagccaaggg gaaggatgac cctggcagat atcgagagga tcgccccact
     1561 ggaggagggc acactgccct ttaacctggc agaggcacag aggcagaagg catccggcga
     1621 ctctgccaga cccgtgctgc tgcaggtggc agagtctgcc tacaggttcg gactgggcag
     1681 cgtggcagga gcagtgggag caaccgccgt gtatcctatc gatctggtga agaccaggat
     1741 gcagaaccag cgcagcacag gctcctttgt gggcgagctg atgtataaga attccttcga
     1801 ctgctttaag aaggtgctga gatacgaggg cttctttggc ctgtataggg gactgctgcc
     1861 tcagctgctg ggagtggcac cagagaaggc catcaagctg accgtgaacg acttcgtgcg
     1921 ggataagttt atgcacaagg atggcagcgt gcctctggcc gccgagatcc tggcaggagg
     1981 atgtgcagga ggatcccagg tcatcttcac caatccactg gagatcgtga agatcaggct
     2041 gcaggtggca ggagagatca ccacaggacc aagggtgtct gccctgagcg tggtgcgcga
     2101 tctgggcttc tttggcatct acaagggcgc caaggcctgc tttctgagag acatcccctt
     2161 ctccgccatc tactttcctt gttatgccca cgtgaaggcc tctttcgcca acgaggacgg
     2221 acaggtgtcc ccaggctctc tgctgctggc aggagcaatc gcaggaatgc cagcagcatc
     2281 tctggtgacc ccagcagatg tgatcaagac aaggctgcag gtggcagcaa gagcaggaca
     2341 gaccacatat tccggcgtga tcgactgctt ccggaagatc ctgagagagg agggaccaaa
     2401 ggccctgtgg aagggagcag gcgccagggt gttccgcagc tcccctcagt ttggcgtgac
     2461 cctgctgaca tacgagctgc tgcagaggtg gttctatatc gattttggcg gcgtgaagcc
     2521 tatgggcagc gagccagtgc ccaagtcccg catcaacctg cctgcaccaa atccagacca
     2581 cgtgggagga tacaagctgg cagtggccac cttcgccgga atcgagaata agtttggcct
     2641 gtatctgcct ctgttcaagc catccgtgtc tacaagcaag gccatcggcg gcggcccacc
     2701 accggtcgga aagtcttacc caactgtgag tgctgattac caggacgccg ttgagaaggc
     2761 gaagaagaag ctcagaggct tcatcgctga gaagagatgc gctcctctaa tgctccgttt
     2821 ggcattccac tctgctggaa cctttgacaa gggcacgaag actggtggac ccttcggaac
     2881 catcaagcac cctgccgaac tggctcacag cgctaacaac ggtcttgaca tcgctgttag
     2941 gcttttggag ccactcaagg cggagttccc tattttgagc tacgccgatt tctaccagtt
     3001 ggctggcgtt gttgccgttg aggtcacggg tggacctaag gttccattcc accctggaag
     3061 agaggacaag cctgagccac caccagaggg tcgcttgccc gatcccacta agggttctga
     3121 ccatttgaga gatgtgtttg gcaaagctat ggggcttact gaccaagata tcgttgctct
     3181 atctgggggt cacactattg gagctgcaca caaggagcgt tctggatttg agggtccctg
     3241 gacctctaat cctcttattt tcgacaactc atacttcacg gagttgttga gtggtgagaa
     3301 ggaaggtctc cttcagctac cttctgacaa ggctcttttg tctgaccctg tattccgccc
     3361 tctcgttgac aaatatgcag cggacgaaga tgccttcttt gctgattacg ctgaggctca
     3421 ccaaaagctt tccgagcttg ggtttgctga tgcctctaga ggcaaaccca taccgaatcc
     3481 cctgctggga ctggatagca caacccagct ttcttgtaca aagttggcat tataaaaaag
     3541 cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt
     3601 tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg
     3661 cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca
     3721 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat
     3781 attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg
     3841 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg
     3901 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
     3961 acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag
     4021 cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca
     4081 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
     4141 gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
     4201 gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
     4261 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt
     4321 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
     4381 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
     4441 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
     4501 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
     4561 atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt
     4621 acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt gagttacgcg
     4681 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4741 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4801 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4861 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4921 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4981 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5041 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5101 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5161 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5221 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5281 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5341 cggttcctgg ccttttgctg gccttttgct cacatgtt
//
