LOCUS       sequence-460058-        5294 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses APEX2-SOPP3 in the mitochondrial matrix.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5294)
  AUTHORS   Qu D, Li Y, Liu Q, Cao B, Cao M, Lin X, Shen C, Zou P, Zhou H, Zhang
            W, Pan W
  TITLE     Photoactivated SOPP3 enables APEX2-mediated proximity labeling with 
            high spatio-temporal resolution in live cells.
  JOURNAL   Cell Res. 2025 Feb;35(2):149-152. doi: 10.1038/s41422-024-01061-9. 
            Epub 2024 Dec 10.
  PUBMED    39653757
REFERENCE   2  (bases 1 to 5294)
  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: 
            "10.1038/s41422-024-01061-9"; journalName: "Cell Res"; date: 
            "2025-02"; volume: "35"; issue: "2"; pages: "149-152"
COMMENT     Sequence Label: COX8mls-Flag-APEX2-SOPP3
FEATURES             Location/Qualifiers
     source          1..5294
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        891..909
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     891..908
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1040..1063
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2202..2336
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2226..2245)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2280..2299
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2449..2469)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2449..2467)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2488..2504)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2488..2504)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2501..2523)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2512..2528
                     /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(2536..2542,2543..2560,2561..2566))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2581..2602
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2719..2736)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2890..3478)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2970..2989)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3649..4440,4441..4509))
                     /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     4272..4291
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4510..4614)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4640..5095)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4777..4798)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4989..5008
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(5166..5294,1..856)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 caattcaata tggcggactt ggcaccatgc caattcaata tggcggacct ggcactgtgc
       61 caactgggga ggggtctact tggcacggtg ccaagtttga ggaggggtct tggccctgtg
      121 ccaagtccgc catattgaat tggcatggtg ccaataatgg cggccatatt ggctatatgc
      181 caggatcaat atataggcaa tatccaatat ggccctatgc caatatggct attggccagg
      241 ttcaatacta tgtattggcc ctatgccata tagtattcca tatatgggtt ttcctattga
      301 cgtagatagc ccctcccaat gggcggtccc atataccata tatggggctt cctaataccg
      361 cccatagcca ctcccccatt gacgtcaatg gtctctatat atggtctttc ctattgacgt
      421 catatgggcg gtcctattga cgtatatggc gcctccccca ttgacgtcaa ttacggtaaa
      481 tggcccgcct ggctcaatgc ccattgacgt caataggacc acccaccatt gacgtcaatg
      541 ggatggctca ttgcccattc atatccgttc tcacgccccc tattgacgtc aatgacggta
      601 aatggcccac ttggcagtac atcaatatct attaatagta acttggcaag tacattacta
      661 ttggaagtac gccagggtac attggcagta ctcccattga cgtcaatggc ggtaaatggc
      721 ccgcgatggc tgccaagtac atccccattg acgtcaatgg ggaggggcaa tgacgcaaat
      781 gggcgttcca ttgacgtaaa tgggcggtag gcgtgcctaa tgggaggtct atataagcaa
      841 tgctcgttta gggaaccgcc attctgcctg gggacgtcgg agcaagcttg atttaggtga
      901 cactatagaa tacaagctac ttgttctttt tgcaggatcc atgtctggac ttctgagggg
      961 actagctcgc gtccgcgccg ctccggttct gcggggatcc acgatcaccc agcgagccaa
     1021 cctcgttacg cgagctagcg actacaagga tgacgatgac aaagaattcg ggaaatcata
     1081 cccaacagtg tccgcagact accaggatgc cgtggagaaa gccaagaaga agctgagagg
     1141 gtttatcgca gaaaagaggt gcgcacctct gatgctgaga ctggctttcc acagcgcagg
     1201 cacctttgac aagggaacca aaacaggcgg accctttgga acaatcaagc accctgctga
     1261 actggcacat agtgccaaca atgggctgga catcgccgtg cggctgctgg aacctctgaa
     1321 agcagagttc ccaattctgt cctacgccga tttttatcag ctggcaggag tggtcgctgt
     1381 ggaggtcact gggggcccca aggtgccttt ccacccagga cgggaggaca aaccagaacc
     1441 acctccagag gggcgcctgc cagatccgac aaagggctcc gaccatctgc gagatgtgtt
     1501 tgggaaagct atgggcctga ctgaccagga tatcgtcgca ctgtctggag ggcacaccat
     1561 tggcgccgct cataaggaaa ggtcaggctt cgagggaccc tggacaagca accccctgat
     1621 tttcgacaat tcttacttta ctgaactgct gagtggagag aaggaagggc tgctgcagct
     1681 gcccagcgac aaagccctgc tgtccgatcc cgtgttcaga cctctggtcg ataagtatgc
     1741 agccgacgag gatgcttttt tcgcagatta cgcagaagca catcagaagc tgtcagaact
     1801 gggatttgcc gacgccaccg gtggcggtgg cggatcgact agtatggaga aaagtttcgt
     1861 gataactgat ccacggctgc cagacaatcc catcatcttc gcatccgatg gcttcctgga
     1921 gctgaccgag tattccagag aggagatcct gggccgcaat ggccgctttc tgcagggacc
     1981 agagacagac caggccacag tgcagaagat tcgcgatgcc attagagatc agcgcgagat
     2041 taccgtgcag ctgataaact acacaaaaag cgggaagaaa ttcttgaacc tcctgaacct
     2101 ccagcccatt agggaccaga agggtgagct ccaggctttc atcggagtgg tgctggatgg
     2161 atagctcgag cctctagaac tatagtgagt cgtattacgt agatccagac atgataagat
     2221 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     2281 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     2341 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaat
     2401 tcgcggccgc ggcgccaatg cattgggccc ggtacccagc ttttgttccc tttagtgagg
     2461 gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     2521 gctcacaatt ccacacaaca tacgagccgg gagcataaag tgtaaagcct ggggtgccta
     2581 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     2641 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2701 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     2761 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2821 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2881 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2941 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3001 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3061 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3121 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3181 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3241 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3301 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3361 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3421 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3481 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3541 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3601 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3661 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3721 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     3781 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     3841 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     3901 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     3961 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4021 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4081 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4141 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4201 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4261 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4321 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4381 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4441 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4501 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4561 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     4621 tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     4681 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     4741 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     4801 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     4861 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     4921 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     4981 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     5041 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat
     5101 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     5161 cgccagtcga ccgccaattc aatatggcgt atatggactc atgccaattc aatatggtgg
     5221 atctggacct gtgccaattc aatatggcgt atatggactc gtgccaattc aatatggtgg
     5281 atctggaccc cagc
//
