LOCUS       sequence-460060-        5273 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses APEX2-SOPP3 in the mammalian nucleus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5273)
  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 5273)
  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: Flag-APEX2-SOPP3-3xNLS
FEATURES             Location/Qualifiers
     source          1..5273
                     /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             944..967
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2072..2092
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2096..2116
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2120..2140
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2181..2315
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2205..2224)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2259..2278
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2428..2448)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2428..2446)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2467..2483)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2467..2483)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2480..2502)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2491..2507
                     /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(2515..2521,2522..2539,2540..2545))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2560..2581
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2698..2715)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2869..3457)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2949..2968)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3628..4419,4420..4488))
                     /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     4251..4270
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4489..4593)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4619..5074)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4756..4777)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4968..4987
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(5145..5273,1..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 atggactaca aggatgacga
      961 tgacaaagaa ttcgggaaat catacccaac agtgtccgca gactaccagg atgccgtgga
     1021 gaaagccaag aagaagctga gagggtttat cgcagaaaag aggtgcgcac ctctgatgct
     1081 gagactggct ttccacagcg caggcacctt tgacaaggga accaaaacag gcggaccctt
     1141 tggaacaatc aagcaccctg ctgaactggc acatagtgcc aacaatgggc tggacatcgc
     1201 cgtgcggctg ctggaacctc tgaaagcaga gttcccaatt ctgtcctacg ccgattttta
     1261 tcagctggca ggagtggtcg ctgtggaggt cactgggggc cccaaggtgc ctttccaccc
     1321 aggacgggag gacaaaccag aaccacctcc agaggggcgc ctgccagatc cgacaaaggg
     1381 ctccgaccat ctgcgagatg tgtttgggaa agctatgggc ctgactgacc aggatatcgt
     1441 cgcactgtct ggagggcaca ccattggcgc cgctcataag gaaaggtcag gcttcgaggg
     1501 accctggaca agcaaccccc tgattttcga caattcttac tttactgaac tgctgagtgg
     1561 agagaaggaa gggctgctgc agctgcccag cgacaaagcc ctgctgtccg atcccgtgtt
     1621 cagacctctg gtcgataagt atgcagccga cgaggatgct tttttcgcag attacgcaga
     1681 agcacatcag aagctgtcag aactgggatt tgccgacgcc accggtggcg gtggcggatc
     1741 gactagtatg gagaaaagtt tcgtgataac tgatccacgg ctgccagaca atcccatcat
     1801 cttcgcatcc gatggcttcc tggagctgac cgagtattcc agagaggaga tcctgggccg
     1861 caatggccgc tttctgcagg gaccagagac agaccaggcc acagtgcaga agattcgcga
     1921 tgccattaga gatcagcgcg agattaccgt gcagctgata aactacacaa aaagcgggaa
     1981 gaaattcttg aacctcctga acctccagcc cattagggac cagaagggtg agctccaggc
     2041 tttcatcgga gtggtgctgg atggaatcga tcctaagaag aagagaaagg tggaccctaa
     2101 gaagaagaga aaggtggacc ctaagaagaa gagaaaggtg tagctcgagc ctctagaact
     2161 atagtgagtc gtattacgta gatccagaca tgataagata cattgatgag tttggacaaa
     2221 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     2281 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     2341 tgtttcaggt tcagggggag gtgtgggagg ttttttaatt cgcggccgcg gcgccaatgc
     2401 attgggcccg gtacccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
     2461 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     2521 acgagccggg agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     2581 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     2641 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     2701 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     2761 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     2821 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2881 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2941 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3001 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3061 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3121 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3181 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3241 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3301 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3361 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3421 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3481 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3541 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3601 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3661 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3721 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     3781 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     3841 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3901 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     3961 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     4021 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     4081 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4141 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4201 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4261 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4321 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4381 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4441 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4501 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4561 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa
     4621 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     4681 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     4741 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     4801 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     4861 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     4921 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     4981 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     5041 cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac
     5101 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagtcgac cgccaattca
     5161 atatggcgta tatggactca tgccaattca atatggtgga tctggacctg tgccaattca
     5221 atatggcgta tatggactcg tgccaattca atatggtgga tctggacccc agc
//
