LOCUS       sequence-213223-        5052 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Theophylline-inducible expression of APEX2 from a mycobacterial 
            shuttle plasmid (integrating).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5052)
  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 5052)
  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: pMV306-ribo-APEX2
FEATURES             Location/Qualifiers
     source          1..5052
                     /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(3121..3140)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3132..3947
                     /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(3205..3224)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4279..4867
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4768..4787
                     /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 tatgccatca tggccgcggt gatcagctag agccgtgaac gacagggcga acgccagccc
     1261 gccgacggcg agggttccga ccgctgcaac tcccggtgca accttgtccc ggtctattct
     1321 cttcactgca ccagctccaa tctggtgtga atgcccctcg tctgttcgcg caggcggggg
     1381 gctctattcg tttgtcagca tcgaaagtag ccagatcagg gatgcgttgc aaccgcgtat
     1441 gcccaggtca gaagagtcgc acaagagttg cagacccctg gaaagaaaaa tggccagagg
     1501 gcgaaaacac cctctgacca gcggagcggg cgacgggaat cgaacccgcg tagctagttt
     1561 ggaagaatgg gtgtctgccg accacatata tgggccggtc aagataggtt tttaccccct
     1621 ctcggctgca tcctctaagt ggaaagaaat tgcaggtcgt agaagcgcgt tgaagcctga
     1681 gagttgcaca ggagttgcaa cccggtagcc ttgttcacga cgagaggaga cctagttggc
     1741 acgtcgcgga tggggatcgc tgaagactca gcgcagcggg aggatccaag cctcatacgt
     1801 caacccgcag gacggtgtga ggtactacgc gctgcagacc tacgacaaca agatggacgc
     1861 cgaagcctgg ctcgcgggcg agaagcggct catcgagatg gagacctgga cccctccaca
     1921 ggaccgggcg aagaaggcag ccgccagcgc catcacgctg gaggagtaca cccggaagtg
     1981 gctcgtggag cgcgacctcg cagacggcac cagggatctg tacagcgggc acgcggagcg
     2041 ccgcatctac ccggtgctag gtgaagtggc ggtcacagag atgacgccag ctctggtgcg
     2101 tgcgtggtgg gccgggatgg gtaggaagca cccgactgcc cgccggcatg cctacaacgt
     2161 cctccgggcg gtgatgaaca cagcggtcga ggacaagctg atcgcagaga acccgtgccg
     2221 gatcgagcag aaggcagccg atgagcgcga cgtagaggcg ctgacgcctg aggagctgga
     2281 catcgtcgcc gctgagatct tcgagcacta ccggatcgcg gcatacatcc tggcgtggac
     2341 gagcctccgg ttcggagagc tgatcgagct tcgccgcaag gacatcgtgg acgacggcat
     2401 gacgatgaag ctccgggtgc gccgtggcgc ttcccgcgtg gggaacaaga tcgtcgttgg
     2461 caacgccaag accgtccggt cgaagcgtcc tgtgacggtt ccgcctcacg tcgcggagat
     2521 gatccgagcg cacatgaagg accgtacgaa gatgaacaag ggccccgagg cattcctggt
     2581 gaccacgacg cagggcaacc ggctgtcgaa gtccgcgttc accaagtcgc tgaagcgtgg
     2641 ctacgccaag atcggtcggc cggaactccg catccacgac ctccgcgctg tcggcgctac
     2701 gttcgccgct caggcaggtg cgacgaccaa ggagctgatg gcccgtctcg gtcacacgac
     2761 tcctaggatg gcgatgaagt accagatggc gtctgaggcc cgcgacgagg ctatcgctga
     2821 ggcgatgtcc aagctggcca agacctcctg aaacgcaaaa agcccccctc ccaaggacac
     2881 tgagtcctaa agaggggggt ttcttgtcag tacgcgaaga accacgcctg gccgcgagcg
     2941 ccagcaccgc cgctctgtgc ggagacctgg gcaccagccc cgccgccgcc aggagcattg
     3001 ccgttcccgc cagctagcaa caaagcgacg ttgtgtctca aaatctctga tgttacattg
     3061 cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata
     3121 caaggggtgt tatgagccat attcaacggg aaacgtcttg ctcgaggccg cgattaaatt
     3181 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag
     3241 gtgcgacaat ctatcgcttg tatgggaagc cccatgcgcc agagttgttt ctgaaacatg
     3301 gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg
     3361 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac
     3421 tcaccactgc gatccccggg aaaacagcat tccaggtatt agaagaatat cctgattcag
     3481 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt
     3541 gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga
     3601 ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac
     3661 aagtctggaa agaaatgcat aatcttttgc cattctcacc ggattcagtc gtcactcatg
     3721 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg
     3781 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg
     3841 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg
     3901 atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta
     3961 attggttgta acactggcag agcattacgc tgacttgacg ggacggcggc tttgttgaat
     4021 aaatcgaact tttgctgagt tgaaggatca gatcacgcat cttcccgaca acgcagaccg
     4081 ttccgtggca aagcaaaagt tcaaaatcac caactggtcc acctacaaca aagctctcat
     4141 caaccgtggc tccctcactt tctggctgga tgatggggcg attcaggcct ggtatgagtc
     4201 agcaacacct tcttcacgag gcagacctca ctagttccac tgagcgtcag accccgtaga
     4261 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     4321 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     4381 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     4441 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4501 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4561 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4621 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc attgagaaag
     4681 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4741 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4801 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4861 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4921 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     4981 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc aacgcgtgcg gccgcggtac
     5041 cagatcttta aa
//
