LOCUS       sequence-221111-        5070 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 5070)
  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 5070)
  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..5070
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..42
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             43..789
                     /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      859..905
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2842..2861)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2853..3668
                     /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(2926..2945)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3999..4587
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4488..4507
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggcaagccca tccccaaccc cctgctgggc ctggacagca ccggaaagtc ttacccaact
       61 gtgagtgctg attaccagga cgccgttgag aaggcgaaga agaagctcag aggcttcatc
      121 gctgagaaga gatgcgctcc tctaatgctc cgtttggcat tccactctgc tggaaccttt
      181 gacaagggca cgaagaccgg tggacccttc ggaaccatca agcaccctgc cgaactggct
      241 cacagcgcta acaacggtct tgacatcgct gttaggcttt tggagccact caaggcggag
      301 ttccctattt tgagctacgc cgatttctac cagttggctg gcgttgttgc cgttgaggtc
      361 acgggtggac ctaaggttcc attccaccct ggaagagagg acaagcctga gccaccacca
      421 gagggtcgct tgcccgatcc cactaagggt tctgaccatt tgagagatgt gtttggcaaa
      481 gctatggggc ttactgacca agatatcgtt gctctatctg ggggtcacac tattggagct
      541 gcacacaagg agcgttctgg atttgagggt ccctggacct ctaatcctct tattttcgac
      601 aactcatact tcacggagtt gttgagtggt gagaaggaag gtctccttca gctaccttct
      661 gacaaggctc ttttgtctga ccctgtattc cgccctctcg ttgataaata tgcagcggac
      721 gaagatgcct tctttgctga ttacgctgag gctcaccaaa agctttccga gcttgggttt
      781 gctgatgcct aatgaagatg gagaccaagc ttatcgatgt cgacgtagtt aactagcgta
      841 cgatcgactg ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
      901 tttatgccat catggccgcg gtgatcagct agagccgtga acgacagggc gaacgccagc
      961 ccgccgacgg cgagggttcc gaccgctgca actcccggtg caaccttgtc ccggtctatt
     1021 ctcttcactg caccagctcc aatctggtgt gaatgcccct cgtctgttcg cgcaggcggg
     1081 gggctctatt cgtttgtcag catcgaaagt agccagatca gggatgcgtt gcaaccgcgt
     1141 atgcccaggt cagaagagtc gcacaagagt tgcagacccc tggaaagaaa aatggccaga
     1201 gggcgaaaac accctctgac cagcggagcg ggcgacggga atcgaacccg cgtagctagt
     1261 ttggaagaat gggtgtctgc cgaccacata tatgggccgg tcaagatagg tttttacccc
     1321 ctctcggctg catcctctaa gtggaaagaa attgcaggtc gtagaagcgc gttgaagcct
     1381 gagagttgca caggagttgc aacccggtag ccttgttcac gacgagagga gacctagttg
     1441 gcacgtcgcg gatggggatc gctgaagact cagcgcagcg ggaggatcca agcctcatac
     1501 gtcaacccgc aggacggtgt gaggtactac gcgctgcaga cctacgacaa caagatggac
     1561 gccgaagcct ggctcgcggg cgagaagcgg ctcatcgaga tggagacctg gacccctcca
     1621 caggaccggg cgaagaaggc agccgccagc gccatcacgc tggaggagta cacccggaag
     1681 tggctcgtgg agcgcgacct cgcagacggc accagggatc tgtacagcgg gcacgcggag
     1741 cgccgcatct acccggtgct aggtgaagtg gcggtcacag agatgacgcc agctctggtg
     1801 cgtgcgtggt gggccgggat gggtaggaag cacccgactg cccgccggca tgcctacaac
     1861 gtcctccggg cggtgatgaa cacagcggtc gaggacaagc tgatcgcaga gaacccgtgc
     1921 cggatcgagc agaaggcagc cgatgagcgc gacgtagagg cgctgacgcc tgaggagctg
     1981 gacatcgtcg ccgctgagat cttcgtgcac taccggatcg cggcatacat cctggcgtgg
     2041 acgagcctcc ggttcggaga gctgatcgag cttcgccgca aggacatcgt ggacgacggc
     2101 atgacgatga agctccgggt gcgccgtggc gcttcccgcg tggggaacaa gatcgtcgtt
     2161 ggcaacgcca agaccgtccg gtcgaagcgt cctgtgacgg ttccgcctca cgtcgcggag
     2221 atgatccgag cgcacatgaa ggaccgtacg aagatgaaca agggccccga ggcattcctg
     2281 gtgaccacga cgcagggcaa ccggctgtcg aagtccgcgt tcaccaagtc gctgaagcgt
     2341 ggctacgcca agatcggtcg gccggaactc cgcatccacg acctccgcgc tgtcggcgct
     2401 acgttcgccg ctcaggcagg tgcgacgacc aaggagctga tggcccgtct cggtcacacg
     2461 actcctagga tggcgatgaa gtaccagatg gcgtctgagg cccgcgacga ggctatcgct
     2521 gaggcgatgt ccaagctggc caagacctcc tgaaacgcaa aaagcccccc tcccaaggac
     2581 actgagtcct aaagaggggg gtttcttgtc agtacgcgaa gaaccacgcc tggccgcgag
     2641 cgccagcacc gccgctctgt gcggagacct gggcaccagc cccgccgccg ccaggagcat
     2701 tgccgttccc gccagctagc cacctgacgt cggggggggg ggaaagccac gttgtgtctc
     2761 aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt
     2821 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt
     2881 gctcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     2941 gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag ccccatgcgc
     3001 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     3061 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     3121 ctcctgatga tgcatggtta ctcaccactg cgatccccgg gaaaacagca ttccaggtat
     3181 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     3241 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
     3301 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
     3361 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taatcttttg ccattctcac
     3421 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     3481 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     3541 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     3601 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
     3661 ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac
     3721 gggacggcgg ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc agatcacgca
     3781 tcttcccgac aacgcagacc gttccgtggc aaagcaaaag ttcaaaatca ccaactggtc
     3841 cacctacaac aaagctctca ccaaccgtgg ctccctcact ttctggctgg atgatggggc
     3901 gattcaggcc tggtatgagt cagcaacacc ttcttcacga ggcagacctc actagttcca
     3961 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4021 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4081 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     4141 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     4201 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     4261 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     4321 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     4381 cagcgtgagc tatgagaaag cgccacgctt cccgagggga gaaaggcgga caggtatccg
     4441 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     4501 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     4561 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     4621 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     4681 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     4741 aacgcgtgcg gccgcggtac cagatcttta aatctagagg tgaccacaac gcgcccgctt
     4801 tgatcgggga cgtctgcggc cgaccattta cgggtcttgt tgtcgttggc ggtcatgggc
     4861 cgaacatact cacccggatc ggagggccga ggacaaggtc gaacgagggg catgacccgg
     4921 tgcggggctt cttgcactcg gcataggcga gtgctaagaa taacgttggc actcgcgacc
     4981 gggatacgac tcactatagg taccggtgat accagcatcg tcttgatgcc cttggcagca
     5041 ccctgctaag gaggcaacaa gatgcatatg
//
