LOCUS       sequence-348175-        5104 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of APEX2 in the periplasm from a codon-optimized gene 
            (single-copy integrating plasmid).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5104)
  AUTHORS   Ahamed M, Jaisinghani N, Li M, Winkeler I, Silva S, Previti ML, 
            Seeliger JC
  TITLE     Optimized APEX2 peroxidase-mediated proximity labeling in fast- and 
            slow-growing mycobacteria.
  JOURNAL   Methods Enzymol. 2022;664:267-289. doi: 10.1016/bs.mie.2021.11.021. 
            Epub 2021 Dec 30.
  PUBMED    35331378
REFERENCE   2  (bases 1 to 5104)
  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: "Methods 
            Enzymol."; date: "2022"; volume: "664:267-289. doi"; pages: " 
            10.1016/bs.mie.2021.11.021. Epub 2021 Dec 30"
COMMENT     Sequence Label: pRiboI-Sec-APEX2m
FEATURES             Location/Qualifiers
     source          1..5104
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      419..428
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     terminator      1271..1317
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3254..3273)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3265..4080
                     /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(3338..3357)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4411..4999
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4900..4919
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcaa cgcgtgcggc
       61 cgcggtacca gatctttaaa tctagaggtg accacaacgc gcccgctttg atcggggacg
      121 tctgcggccg accatttacg ggtcttgttg tcgttggcgg tcatgggccg aacatactca
      181 cccggatcgg agggccgagg acaaggtcga acgaggggca tgacccggtg cggggcttct
      241 tgcactcggc ataggcgagt gctaagaata acgttggcac tcgcgaccgg gatacgactc
      301 actataggta ccggtgatac cagcatcgtc ttgatgccct tggcagcacc ctgctaagga
      361 ggcaacaaga tgcatatgaa gctcaccaca atgatcaaga cggcagtagc ggtcgtggcc
      421 atggcggcca tcgcgacctt tgcggcaccg gtcgcgttgg ctggatccgg caagagctac
      481 ccgaccgtca gcgccgacta ccaggacgcc gtggagaagg ccaagaagaa gctgcgcggc
      541 ttcatcgccg agaagcggtg cgccccgctg atgctgcgcc tggcgttcca cagcgcgggc
      601 accttcgaca agggcaccaa gaccggcggc ccgttcggca ccatcaagca cccggcggag
      661 ctggcgcaca gcgccaacaa cggcctggac atcgcggtcc ggctgctgga gccgctgaag
      721 gcggagttcc cgatcctgag ctacgccgac ttctaccagc tggcgggcgt cgtggccgtc
      781 gaggtgaccg gcggcccgaa ggtgccgttc cacccgggcc gggaggacaa gccggagccg
      841 ccgccggagg gccgcctgcc ggacccgacc aagggctcgg accacctgcg ggacgtcttc
      901 ggcaaggcga tgggcctgac cgaccaggac atcgtggccc tgagcggcgg ccacaccatc
      961 ggcgcggccc acaaggagcg cagcggcttc gagggcccgt ggaccagcaa cccgctgatc
     1021 ttcgacaact cgtacttcac cgagctgctg agcggcgaga aggaaggcct gctgcagctg
     1081 ccgagcgaca aggcgctgct gagcgacccg gtcttccggc ccctggtgga caagtacgcg
     1141 gccgacgagg acgccttctt cgccgactac gccgaggccc accagaagct gtccgagctg
     1201 ggcttcgccg acgccaagct ttgaatcgat gtcgacgtag ttaactagcg tacgatcgac
     1261 tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatgcc
     1321 atcatggccg cggtgatcag ctagagccgt gaacgacagg gcgaacgcca gcccgccgac
     1381 ggcgagggtt ccgaccgctg caactcccgg tgcaaccttg tcccggtcta ttctcttcac
     1441 tgcaccagct ccaatctggt gtgaatgccc ctcgtctgtt cgcgcaggcg gggggctcta
     1501 ttcgtttgtc agcatcgaaa gtagccagat cagggatgcg ttgcaaccgc gtatgcccag
     1561 gtcagaagag tcgcacaaga gttgcagacc cctggaaaga aaaatggcca gagggcgaaa
     1621 acaccctctg accagcggag cgggcgacgg gaatcgaacc cgcgtagcta gtttggaaga
     1681 atgggtgtct gccgaccaca tatatgggcc ggtcaagata ggtttttacc ccctctcggc
     1741 tgcatcctct aagtggaaag aaattgcagg tcgtagaagc gcgttgaagc ctgagagttg
     1801 cacaggagtt gcaacccggt agccttgttc acgacgagag gagacctagt tggcacgtcg
     1861 cggatgggga tcgctgaaga ctcagcgcag cgggaggatc caagcctcat acgtcaaccc
     1921 gcaggacggt gtgaggtact acgcgctgca gacctacgac aacaagatgg acgccgaagc
     1981 ctggctcgcg ggcgagaagc ggctcatcga gatggagacc tggacccctc cacaggaccg
     2041 ggcgaagaag gcagccgcca gcgccatcac gctggaggag tacacccgga agtggctcgt
     2101 ggagcgcgac ctcgcagacg gcaccaggga tctgtacagc gggcacgcgg agcgccgcat
     2161 ctacccggtg ctaggtgaag tggcggtcac agagatgacg ccagctctgg tgcgtgcgtg
     2221 gtgggccggg atgggtagga agcacccgac tgcccgccgg catgcctaca acgtcctccg
     2281 ggcggtgatg aacacagcgg tcgaggacaa gctgatcgca gagaacccgt gccggatcga
     2341 gcagaaggca gccgatgagc gcgacgtaga ggcgctgacg cctgaggagc tggacatcgt
     2401 cgccgctgag atcttcgtgc actaccggat cgcggcatac atcctggcgt ggacgagcct
     2461 ccggttcgga gagctgatcg agcttcgccg caaggacatc gtggacgacg gcatgacgat
     2521 gaagctccgg gtgcgccgtg gcgcttcccg cgtggggaac aagatcgtcg ttggcaacgc
     2581 caagaccgtc cggtcgaagc gtcctgtgac ggttccgcct cacgtcgcgg agatgatccg
     2641 agcgcacatg aaggaccgta cgaagatgaa caagggcccc gaggcattcc tggtgaccac
     2701 gacgcagggc aaccggctgt cgaagtccgc gttcaccaag tcgctgaagc gtggctacgc
     2761 caagatcggt cggccggaac tccgcatcca cgacctccgc gctgtcggcg ctacgttcgc
     2821 cgctcaggca ggtgcgacga ccaaggagct gatggcccgt ctcggtcaca cgactcctag
     2881 gatggcgatg aagtaccaga tggcgtctga ggcccgcgac gaggctatcg ctgaggcgat
     2941 gtccaagctg gccaagacct cctgaaacgc aaaaagcccc cctcccaagg acactgagtc
     3001 ctaaagaggg gggtttcttg tcagtacgcg aagaaccacg cctggccgcg agcgccagca
     3061 ccgccgctct gtgcggagac ctgggcacca gccccgccgc cgccaggagc attgccgttc
     3121 ccgccagcta gccacctgac gtcggggggg ggggaaagcc acgttgtgtc tcaaaatctc
     3181 tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac
     3241 ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc ttgctcgagg
     3301 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
     3361 gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agccccatgc gccagagttg
     3421 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
     3481 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
     3541 gatgcatggt tactcaccac tgcgatcccc gggaaaacag cattccaggt attagaagaa
     3601 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
     3661 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg
     3721 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
     3781 tggcctgttg aacaagtctg gaaagaaatg cataatcttt tgccattctc accggattca
     3841 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     3901 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     3961 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     4021 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     4081 tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc
     4141 ggctttgttg aataaatcga acttttgctg agttgaagga tcagatcacg catcttcccg
     4201 acaacgcaga ccgttccgtg gcaaagcaaa agttcaaaat caccaactgg tccacctaca
     4261 acaaagctct caccaaccgt ggctccctca ctttctggct ggatgatggg gcgattcagg
     4321 cctggtatga gtcagcaaca ccttcttcac gaggcagacc tcactagttc catgagcgtc
     4381 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4441 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4501 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     4561 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4621 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4681 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4741 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4801 gctatgagaa agcgccacgc ttcccgaggg gagaaaggcg gacaggtatc cggtaagcgg
     4861 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4921 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4981 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5041 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     5101 tacc
//
