LOCUS       sequence-73408-e        2673 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses a tandem GFP-APEX2 fusion to the C-terminus of connexin43 
            in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2673)
  AUTHORS   Lam SS, Martell JD, Kamer KJ, Deerinck TJ, Ellisman MH, Mootha VK, 
            Ting AY
  TITLE     Directed evolution of APEX2 for electron microscopy and proximity 
            labeling.
  JOURNAL   Nat Methods. 2014 Nov 24. doi: 10.1038/nmeth.3179.
  PUBMED    25419960
REFERENCE   2  (bases 1 to 2673)
  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: "Nat 
            Methods. 2014 Nov 24. doi: 10.1038/nmeth.3179."
COMMENT     Sequence Label: pcDNA3 Connexin43-GFP-APEX2
FEATURES             Location/Qualifiers
     source          1..2673
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             join(1168..1170,1171..1173,1174..1884)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1213..1234)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1474..1493)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1821..1842
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1924..2670
                     /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"
ORIGIN
        1 atgggtgact ggagtgcctt ggggaaatta ctggacaagg tccaagccta ctccaccgct
       61 ggagggaagg tgtggctgtc agtgctcttc atattcagaa tcctgctcct ggggacagct
      121 gttgagtcag cttggggtga tgaacagtct gcctttcgct gtaacactca acaacctggc
      181 tgcgaaaacg tctgctatga caagtccttc cccatctctc acgtgcgctt ctgggtcctt
      241 cagatcatat tcgtgtctgt gcccacactc ctgtacttgg cccatgtgtt ctatgtgatg
      301 aggaaggaag agaagctaaa caagaaagaa gaggagctca aagtggccca gactgacggg
      361 gtcaacgtgg agatgcacct gaagcagatt gaaatcaaga agttcaagta cgggattgaa
      421 gagcacggca aggtgaaaat gaggggcggc ttgctgagaa cctacatcat cagcatcctc
      481 ttcaagtctg tcttcgaggt ggccttcctg ctcatccagt ggtacatcta tgggttcagc
      541 ttgagcgcgg tctacacctg caagagagat ccctgccccc accaggtaga ctgcttcctc
      601 tcacgtccca cggagaaaac catcttcatc atcttcatgc tggtggtgtc cttggtgtct
      661 ctcgctttga acatcattga gctcttctac gtcttcttca agggcgttaa ggatcgcgtg
      721 aagggaagaa gcgatcctta ccacgccacc actggcccac tgagcccatc aaaagactgc
      781 ggatctccaa aatacgccta cttcaatggc tgctcctcac caacggctcc actctcgcct
      841 atgtctcctc ctgggtacaa gctggttact ggtgacagaa acaattcctc gtgccgcaat
      901 tacaacaagc aagctagcga gcaaaactgg gcgaactaca gcgcagagca aaatcgcatg
      961 gggcaggccg gaagcaccat ctccaactcg cacgcccagc cgttcgattt ccccgacgac
     1021 aaccagaatg ccaaaaaagt tgctgctgga catgaactcc agccattagc catcgtggac
     1081 caacgacctt ccagcagagc cagcagccgc gccagcagca ggcctcggcc tgatgacctg
     1141 gagattggat ccgggggtgg tgggtctatg gtgagcaagg gcgaggagct gttcaccggg
     1201 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     1261 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     1321 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     1381 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     1441 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     1501 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     1561 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     1621 tatatcatgg ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac
     1681 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     1741 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
     1801 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     1861 ctcggcatgg acgagctgta caaggggcgc gccaagggct cgggctcgac ctcgggctcg
     1921 ggcggaaagt cttacccaac tgtgagtgct gattaccagg acgccgttga gaaggcgaag
     1981 aagaagctca gaggcttcat cgctgagaag agatgcgctc ctctaatgct ccgtttggca
     2041 ttccactctg ctggaacctt tgacaagggc acgaagaccg gtggaccctt cggaaccatc
     2101 aagcaccctg ccgaactggc tcacagcgct aacaacggtc ttgacatcgc tgttaggctt
     2161 ttggagccac tcaaggcgga gttccctatt ttgagctacg ccgatttcta ccagttggct
     2221 ggcgttgttg ccgttgaggt cacgggtgga cctaaggttc cattccaccc tggaagagag
     2281 gacaagcctg agccaccacc agagggtcgc ttgcccgatc ccactaaggg ttctgaccat
     2341 ttgagagatg tgtttggcaa agctatgggg cttactgacc aagatatcgt tgctctatct
     2401 gggggtcaca ctattggagc tgcacacaag gagcgttctg gatttgaggg tccctggacc
     2461 tctaatcctc ttattttcga caactcatac ttcacggagt tgttgagtgg tgagaaggaa
     2521 ggtctccttc agctaccttc tgacaaggct cttttgtctg accctgtatt ccgccctctc
     2581 gttgacaaat atgcagcgga cgaagatgcc ttctttgctg attacgctga ggctcaccaa
     2641 aagctttccg agcttgggtt tgctgatgcc taa
//
