LOCUS       sequence-213229-        5136 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Theophylline-inducible expression of APEX2 from a mycobacterial 
            shuttle plasmid (episomal).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5136)
  AUTHORS   Ganapathy US, Bai L, Wei L, Eckartt KA, Lett CM, Previti ML, Carrico
            IS, Seeliger JC
  TITLE     Compartment-Specific Labeling of Bacterial Periplasmic Proteins by 
            Peroxidase-Mediated Biotinylation.
  JOURNAL   ACS Infect Dis. 2018 Apr 30. doi: 10.1021/acsinfecdis.8b00044.
  PUBMED    29708735
REFERENCE   2  (bases 1 to 5136)
  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: "ACS Infect 
            Dis. 2018 Apr 30. doi: 10.1021/acsinfecdis.8b00044."
COMMENT     Sequence Label: pRibo-APEX2
FEATURES             Location/Qualifiers
     source          1..5136
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             299..340
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             341..1087
                     /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"
     terminator      1157..1203
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1349..1368)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1360..2175
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPHAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHNLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1433..1452)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2507..3095
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2996..3015
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tctagaggtg accacaacgc gcccgctttg atcggggacg tctgcggccg accatttacg
       61 ggtcttgttg tcgttggcgg tcatgggccg aacatactca cccggatcgg agggccgagg
      121 acaaggtcga acgaggggca tgacccggtg cggggcttct tgcactcggc ataggcgagt
      181 gctaagaata acgttggcac tcgcgaccgg gatacgactc actataggta ccggtgatac
      241 cagcatcgtc ttgatgccct tggcagcacc ctgctaagga ggcaacaaga tgcatatggg
      301 caagcccatc cccaaccccc tgctgggcct ggacagcacc ggaaagtctt acccaactgt
      361 gagtgctgat taccaggacg ccgttgagaa ggcgaagaag aagctcagag gcttcatcgc
      421 tgagaagaga tgcgctcctc taatgctccg tttggcattc cactctgctg gaacctttga
      481 caagggcacg aagaccggtg gacccttcgg aaccatcaag caccctgccg aactggctca
      541 cagcgctaac aacggtcttg acatcgctgt taggcttttg gagccactca aggcggagtt
      601 ccctattttg agctacgccg atttctacca gttggctggc gttgttgccg ttgaggtcac
      661 gggtggacct aaggttccat tccaccctgg aagagaggac aagcctgagc caccaccaga
      721 gggtcgcttg cccgatccca ctaagggttc tgaccatttg agagatgtgt ttggcaaagc
      781 tatggggctt actgaccaag atatcgttgc tctatctggg ggtcacacta ttggagctgc
      841 acacaaggag cgttctggat ttgagggtcc ctggacctct aatcctctta ttttcgacaa
      901 ctcatacttc acggagttgt tgagtggtga gaaggaaggt ctccttcagc taccttctga
      961 caaggctctt ttgtctgacc ctgtattccg ccctctcgtt gataaatatg cagcggacga
     1021 agatgccttc tttgctgatt acgctgaggc tcaccaaaag ctttccgagc ttgggtttgc
     1081 tgatgcctaa tgaagatgga gaccaagctt atcgatgtcg acgtagttaa ctagcgtacg
     1141 atcgactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     1201 tatctgttgt ttgtccggcc atcatggccg cggtgatcag ctagccaaca aagcgacgtt
     1261 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata
     1321 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
     1381 acgtcttgct cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
     1441 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc
     1501 catgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
     1561 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt
     1621 atccgtactc ctgatgatgc atggttactc accactgcga tccccgggaa aacagcattc
     1681 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc
     1741 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt
     1801 cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat
     1861 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa tcttttgcca
     1921 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     1981 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
     2041 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     2101 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc
     2161 gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg
     2221 acttgacggg acggcggctt tgttgaataa atcgaacttt tgctgagttg aaggatcaga
     2281 tcacgcatct tcccgacaac gcagaccgtt ccgtggcaaa gcaaaagttc aaaatcacca
     2341 actggtccac ctacaacaaa gctctcatca accgtggctc cctcactttc tggctggatg
     2401 atggggcgat tcaggcctgg tatgagtcag caacaccttc ttcacgaggc agacctcact
     2461 agttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     2521 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     2581 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     2641 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     2701 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     2761 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     2821 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     2881 gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     2941 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3001 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3061 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3121 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     3181 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     3241 ccgagcgcaa cgcgtgagcc caccagctcc gtaagttcgg gtgctgtgtg gctcgtaccc
     3301 gcgcattcag gcggcagggg gtctaacggg tctaaggcgg cgtgtacggc cgccacagcg
     3361 gctcttagcg gcccggaaac gtcctcgaaa cgacgcatgt gttcctcctg gttggtacag
     3421 gtggttgggg gtgctcggct gtcgctggtg tttcatcatc agggctcgac gggagagcgg
     3481 gggagtgtgc agttgtgggg tggcccctca gcgaaatatc tgacttggag ctcgtgtcgg
     3541 accatacacc ggtgattaat cgtggtttat tatcaagcgt gagccacgtc gccgacgaat
     3601 ttgagcagct ctggctgccg tactggtccc tggcaagcga cgatctgctc gaggggatct
     3661 accgccaaag ccgcgcgtcg gccctaggcc gccggtacat cgaggcgaac ccaacagcgc
     3721 tggcaaacct gctggtcgtg gacgtagacc atccagacgc agcgctccga gcgctcagcg
     3781 cccgggggtc ccatccgctg cccaacgcga tcgtgggcaa tcgcgccaac ggccacgcac
     3841 acgcagtgtg ggcactcaac gcccctgttc cacgcaccga atacgcgcgg cgtaagccgc
     3901 gccgtcgatg gcgaccgcag ttactcaggc ctcatgacca aaaaccccgg ccacatcgcc
     3961 tgggaaacgg aatggctcca ctcagatctc tacacactca gccacatcga ggccgagctc
     4021 ggcgcgaaca tgccaccgcc gcgctggcgt cagcagacca cgtacaaagc ggctccgacg
     4081 ccgctagggc ggaattgcgc actgttcgat tccgtcaggt tgtgggccta tcttcccgcc
     4141 ctcatgcgga tctacctgcc gacccggaac gtggacggac tcggccgcgc gatctatgcc
     4201 gagtgccacg cgcgaaacgc cgaatttccg tgcaacgacg tgtgtcccgg accgctaccg
     4261 gacagcgagg tccgcgccat cgccaacagc atttggcgtt ggatcacaac caagtcgcgc
     4321 atttgggcgg acgggatcgt ggtctacgag gccacactca gtgcgcgcca tgcggccatc
     4381 tcgcggaagg gcgcagcagc gcgcacggcg gcgagcacag ttgcgcggcg cgcaaagtcc
     4441 gcgtcagcca tggaggcatt gctatgagcg acggctacag cgacggctac agcgacggct
     4501 acaactggca gccgactgtc cgcaaaaagc ggcgcgtgac cgccgccgaa ggcgctcgaa
     4561 tcaccggact atccgaacgc cacgtcgtcc ggctcgtggc gcaggaacgc agcgagtggt
     4621 tcgccgagca ggctgcacgc cgcgaacgca tccgcgccta tcacgacgac gagggccact
     4681 cttggccgca aacggccaaa catttcgggc tgcatctgga caccgttaag cgactcggct
     4741 atcgggcgag gaaagagcgt gcggcagaac aggaagcggc tcaaaaggcc cacaacgaag
     4801 ccgacaatcc accgctgttc taacgcaatt ggggagcggg tgtcgcgggg gttccgtggg
     4861 gggttccgtt gcaacgggtc ggacaggtaa aagtcctggt agacgctagt tttctggttt
     4921 gggccatgcc tgtctcgttg cgtgtttcgt tgcgtccgtt ttgaatacca gccagacgag
     4981 acggggttct acgaatcttg gtcgatacca agccatttcc gctgaatatc gtggagctca
     5041 ccgccagaat cggtggttgt ggtgatgtac gtggcgaact ccgttgtagt gcttgtggtg
     5101 gcatccgtgg cgcggccgcg gtaccagatc tttaaa
//
