LOCUS       sequence-460059-        5303 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses APEX2-SOPP3 targeted to outer mitochondrial membrane.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5303)
  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 5303)
  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: OMM-Flag-APEX2-SOPP3
FEATURES             Location/Qualifiers
     source          1..5303
                     /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             1049..1072
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2211..2345
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2235..2254)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2289..2308
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2458..2478)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2458..2476)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2497..2513)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2497..2513)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2510..2532)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2521..2537
                     /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(2545..2551,2552..2569,2570..2575))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2590..2611
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2728..2745)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2899..3487)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2979..2998)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3658..4449,4450..4518))
                     /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     4281..4300
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4519..4623)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4649..5104)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4786..4807)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4998..5017
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(5175..5303,1..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 atgatgggcg gcagcagcag
      961 caggatcgcg gctggtttgg gcgcggcgct cttcgtcggt tactgcattt atttcgacag
     1021 gaagagacgg agtgacccga acgctagcga ctacaaggat gacgatgaca aagaattcgg
     1081 gaaatcatac ccaacagtgt ccgcagacta ccaggatgcc gtggagaaag ccaagaagaa
     1141 gctgagaggg tttatcgcag aaaagaggtg cgcacctctg atgctgagac tggctttcca
     1201 cagcgcaggc acctttgaca agggaaccaa aacaggcgga ccctttggaa caatcaagca
     1261 ccctgctgaa ctggcacata gtgccaacaa tgggctggac atcgccgtgc ggctgctgga
     1321 acctctgaaa gcagagttcc caattctgtc ctacgccgat ttttatcagc tggcaggagt
     1381 ggtcgctgtg gaggtcactg ggggccccaa ggtgcctttc cacccaggac gggaggacaa
     1441 accagaacca cctccagagg ggcgcctgcc agatccgaca aagggctccg accatctgcg
     1501 agatgtgttt gggaaagcta tgggcctgac tgaccaggat atcgtcgcac tgtctggagg
     1561 gcacaccatt ggcgccgctc ataaggaaag gtcaggcttc gagggaccct ggacaagcaa
     1621 ccccctgatt ttcgacaatt cttactttac tgaactgctg agtggagaga aggaagggct
     1681 gctgcagctg cccagcgaca aagccctgct gtccgatccc gtgttcagac ctctggtcga
     1741 taagtatgca gccgacgagg atgctttttt cgcagattac gcagaagcac atcagaagct
     1801 gtcagaactg ggatttgccg acgccaccgg tggcggtggc ggatcgacta gtatggagaa
     1861 aagtttcgtg ataactgatc cacggctgcc agacaatccc atcatcttcg catccgatgg
     1921 cttcctggag ctgaccgagt attccagaga ggagatcctg ggccgcaatg gccgctttct
     1981 gcagggacca gagacagacc aggccacagt gcagaagatt cgcgatgcca ttagagatca
     2041 gcgcgagatt accgtgcagc tgataaacta cacaaaaagc gggaagaaat tcttgaacct
     2101 cctgaacctc cagcccatta gggaccagaa gggtgagctc caggctttca tcggagtggt
     2161 gctggatgga tagctcgagc ctctagaact atagtgagtc gtattacgta gatccagaca
     2221 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     2281 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     2341 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     2401 ttttttaatt cgcggccgcg gcgccaatgc attgggcccg gtacccagct tttgttccct
     2461 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     2521 ttgttatccg ctcacaattc cacacaacat acgagccggg agcataaagt gtaaagcctg
     2581 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     2641 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2701 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2761 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2821 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2881 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2941 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3001 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3061 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3121 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3181 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3241 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3301 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3361 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3421 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3481 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3541 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3601 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3661 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3721 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3781 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3841 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3901 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3961 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4021 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4081 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4141 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4201 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4261 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4321 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4381 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4441 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4501 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4561 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4621 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc
     4681 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     4741 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     4801 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     4861 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     4921 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     4981 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     5041 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     5101 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     5161 tcgctattac gccagtcgac cgccaattca atatggcgta tatggactca tgccaattca
     5221 atatggtgga tctggacctg tgccaattca atatggcgta tatggactcg tgccaattca
     5281 atatggtgga tctggacccc agc
//
