LOCUS       sequence-460062-        5492 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresse APEX2-SOPP3 in ER in mammalian cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5492)
  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 5492)
  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: Sec61-beta-Flag-APEX2-SOPP3
FEATURES             Location/Qualifiers
     source          1..5492
                     /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             1238..1261
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2400..2534
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2424..2443)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2478..2497
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2647..2667)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2647..2665)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2686..2702)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2686..2702)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2699..2721)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2710..2726
                     /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(2734..2740,2741..2758,2759..2764))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2779..2800
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2917..2934)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3088..3676)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3168..3187)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3847..4638,4639..4707))
                     /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     4470..4489
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4708..4812)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4838..5293)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4975..4996)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5187..5206
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(5364..5492,1..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 atgcctggac ccgcagctag
      961 tgcaacaaat gttggtgcct ccagccgttc ccccagtaag acggtggctc cccgcactgc
     1021 tggtacctca gccagacaaa ggaaagccac aagcagcagt gcacgcagcg gaggcagatc
     1081 cacagcttct gcaggcacag gaggaatgtg gcgcttttac actgaagatt caccagggct
     1141 taaagttggc ccagttccag ttttggtgat gagtctgctg tttatcgcat ctgtcttcat
     1201 gctgcacatc tggggaaagt acacccgctc cgctagcgac tacaaggatg acgatgacaa
     1261 agaattcggg aaatcatacc caacagtgtc cgcagactac caggatgccg tggagaaagc
     1321 caagaagaag ctgagagggt ttatcgcaga aaagaggtgc gcacctctga tgctgagact
     1381 ggctttccac agcgcaggca cctttgacaa gggaaccaaa acaggcggac cctttggaac
     1441 aatcaagcac cctgctgaac tggcacatag tgccaacaat gggctggaca tcgccgtgcg
     1501 gctgctggaa cctctgaaag cagagttccc aattctgtcc tacgccgatt tttatcagct
     1561 ggcaggagtg gtcgctgtgg aggtcactgg gggccccaag gtgcctttcc acccaggacg
     1621 ggaggacaaa ccagaaccac ctccagaggg gcgcctgcca gatccgacaa agggctccga
     1681 ccatctgcga gatgtgtttg ggaaagctat gggcctgact gaccaggata tcgtcgcact
     1741 gtctggaggg cacaccattg gcgccgctca taaggaaagg tcaggcttcg agggaccctg
     1801 gacaagcaac cccctgattt tcgacaattc ttactttact gaactgctga gtggagagaa
     1861 ggaagggctg ctgcagctgc ccagcgacaa agccctgctg tccgatcccg tgttcagacc
     1921 tctggtcgat aagtatgcag ccgacgagga tgcttttttc gcagattacg cagaagcaca
     1981 tcagaagctg tcagaactgg gatttgccga cgccaccggt ggcggtggcg gatcgactag
     2041 tatggagaaa agtttcgtga taactgatcc acggctgcca gacaatccca tcatcttcgc
     2101 atccgatggc ttcctggagc tgaccgagta ttccagagag gagatcctgg gccgcaatgg
     2161 ccgctttctg cagggaccag agacagacca ggccacagtg cagaagattc gcgatgccat
     2221 tagagatcag cgcgagatta ccgtgcagct gataaactac acaaaaagcg ggaagaaatt
     2281 cttgaacctc ctgaacctcc agcccattag ggaccagaag ggtgagctcc aggctttcat
     2341 cggagtggtg ctggatggat agctcgagcc tctagaacta tagtgagtcg tattacgtag
     2401 atccagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     2461 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     2521 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     2581 tgtgggaggt tttttaattc gcggccgcgg cgccaatgca ttgggcccgg tacccagctt
     2641 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
     2701 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggga gcataaagtg
     2761 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     2821 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     2881 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2941 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3001 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3061 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3121 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3181 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3241 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3301 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3361 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3421 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3481 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     3541 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3601 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3661 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3721 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3781 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3841 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3901 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3961 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4021 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4081 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4141 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4201 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4261 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4321 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4381 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4441 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4501 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4561 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4621 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4681 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4741 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4801 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg
     4861 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     4921 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     4981 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     5041 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     5101 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     5161 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     5221 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     5281 ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     5341 cgggcctctt cgctattacg ccagtcgacc gccaattcaa tatggcgtat atggactcat
     5401 gccaattcaa tatggtggat ctggacctgt gccaattcaa tatggcgtat atggactcgt
     5461 gccaattcaa tatggtggat ctggacccca gc
//
