LOCUS       sequence-348177-        5281 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of APEX2 in the periplasm from a codon-optimized gene 
            (multi-copy episomal plasmid).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5281)
  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 5281)
  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: pRibo-Sec-APEX2m
FEATURES             Location/Qualifiers
     source          1..5281
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      394..403
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1267..1290
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      1328..1374
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1524..1543)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1535..2350
                     /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(1608..1627)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2681..3269
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3170..3189
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttgtagtg cctgtggtgg catccgtggc gcggccgcgg taccagatct ttaaatctag
       61 aggtgaccac aacgcgcccg ctttgatcgg ggacgtctgc ggccgaccat ttacgggtct
      121 tgttgtcgtt ggcggtcatg ggccgaacat actcacccgg atcggagggc cgaggacaag
      181 gtcgaacgag gggcatgacc cggtgcgggg cttcttgcac tcggcatagg cgagtgctaa
      241 gaataacgtt ggcactcgcg accgggatac gactcactat aggtaccggt gataccagca
      301 tcgtcttgat gcccttggca gcaccctgct aaggaggcaa caagatgcat atgaagctca
      361 ccacaatgat caagacggca gtagcggtcg tggccatggc ggccatcgcg acctttgcgg
      421 caccggtcgc gttggctgga tccggcaaga gctacccgac cgtcagcgcc gactaccagg
      481 acgccgtgga gaaggccaag aagaagctgc gcggcttcat cgccgagaag cggtgcgccc
      541 cgctgatgct gcgcctggcg ttccacagcg cgggcacctt cgacaagggc accaagaccg
      601 gcggcccgtt cggcaccatc aagcacccgg cggagctggc gcacagcgcc aacaacggcc
      661 tggacatcgc ggtccggctg ctggagccgc tgaaggcgga gttcccgatc ctgagctacg
      721 ccgacttcta ccagctggcg ggcgtcgtgg ccgtcgaggt gaccggcggc ccgaaggtgc
      781 cgttccaccc gggccgggag gacaagccgg agccgccgcc ggagggccgc ctgccggacc
      841 cgaccaaggg ctcggaccac ctgcgggacg tcttcggcaa ggcgatgggc ctgaccgacc
      901 aggacatcgt ggccctgagc ggcggccaca ccatcggcgc ggcccacaag gagcgcagcg
      961 gcttcgaggg cccgtggacc agcaacccgc tgatcttcga caactcgtac ttcaccgagc
     1021 tgctgagcgg cgagaaggaa ggcctgctgc agctgccgag cgacaaggcg ctgctgagcg
     1081 acccggtctt ccggcccctg gtggacaagt acgcggccga cgaggacgcc ttcttcgccg
     1141 actacgccga ggcccaccag aagctgtccg agctgggctt cgccgacgcc aagctttgaa
     1201 tcgatacatc aggaattcgc tagcgactac aaggacgacg acggcgatta taaagatcat
     1261 gacatcgact acaaggacga cgacgacaag taggtagtta actagcgtac gatcgactgc
     1321 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatgccatc
     1381 atggccgcgg tgatcagcta gccacctgac gtcggggggg ggggaaagcc acgttgtgtc
     1441 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     1501 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     1561 ttgctcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     1621 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agccccatgc
     1681 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     1741 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     1801 tactcctgat gatgcatggt tactcaccac tgcgatcccc gggaaaacag cattccaggt
     1861 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     1921 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     1981 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     2041 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataatcttt tgccattctc
     2101 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     2161 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     2221 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     2281 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     2341 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     2401 acgggacggc ggctttgttg aataaatcga acttttgctg agttgaagga tcagatcacg
     2461 catcttcccg acaacgcaga ccgttccgtg gcaaagcaaa agttcaaaat caccaactgg
     2521 tccacctaca acaaagctct caccaaccgt ggctccctca ctttctggct ggatgatggg
     2581 gcgattcagg cctggtatga gtcagcaaca ccttcttcac gaggcagacc tcactagttc
     2641 catgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     2701 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     2761 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     2821 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2881 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2941 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3001 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3061 tacagcgtga gctatgagaa agcgccacgc ttcccgaggg gagaaaggcg gacaggtatc
     3121 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3181 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3241 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3301 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     3361 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     3421 gcaacgcgtg agcccaccag ctccgtaagt tcgggcgctg tgtggctcgt acccgcgcat
     3481 tcaggcggca gggggtctaa cgggtctaag gcggcgtgta cggccgccac agcggctctc
     3541 agcggcccgg aaacgtcctc gaaacgacgc atgtgttcct cctggttggt acaggtggtt
     3601 gggggtgctc ggctgtcgct ggtgttccac caccagggct cgacgggaga gcgggggagt
     3661 gtgcagttgt ggggtggccc ctcagcgaaa tatctgactt ggagctcgtg tcggaccata
     3721 caccggtgat taatcgtggt ctactaccaa gcgtgagcca cgtcgccgac gaatttgagc
     3781 agctctggct gccgtactgg ccgctggcaa gcgacgatct gctcgagggg atctaccgcc
     3841 aaagccgcgc gtcggcccta ggccgccggt acatcgaggc gaacccaaca gcgctggcaa
     3901 acctgctggt cgtggacgta gaccatccag acgcagcgct ccgagcgctc agcgcccggg
     3961 ggtcccatcc gctgcccaac gcgatcgtgg gcaatcgcgc caacggccac gcacacgcag
     4021 tgtgggcact caacgcccct gttccacgca ccgaatacgc gcggcgtaag ccgctcgcat
     4081 acatggcggc gtgcgccgaa ggccttcggc gcgccgtcga tggcgaccgc agttactcag
     4141 gcctcatgac caaaaacccc ggccacatcg cctgggaaac ggaatggctc cactcagatc
     4201 tctacacact cagccacatc gaggccgagc tcggcgcgaa catgccaccg ccgcgctggc
     4261 gtcagcagac cacgtacaaa gcggctccga cgccgctagg gcggaattgc gcactgttcg
     4321 attccgtcag gttgtgggcc tatcgtcccg ccctcatgcg gatctacctg ccgacccgga
     4381 acgtggacgg actcggccgc gcgatctatg ccgagtgcca cgcgcgaaac gccgaatttc
     4441 cgtgcaacga cgtgtgtccc ggaccgctac cggacagcga ggtccgcgcc atcgccaaca
     4501 gcatttggcg ttggatcaca accaagtcgc gcatttgggc ggacgggatc gtggtctacg
     4561 aggccacact cagtgcgcgc cagtcggcca tctcgcggaa gggcgcagca gcgcgcacgg
     4621 cggcgagcac agttgcgcgg cgcgcaaagt ccgcgtcagc catggaggca ttgctatgag
     4681 cgacggctac agcgacggct acaaccggca gccgactgtc cgcaaaaagc ggcgcgtgac
     4741 cgccgccgaa ggcgctcgaa tcaccggact atccgaacgc cacgtcgtcc ggctcgtggc
     4801 gcaggaacgc agcgagtggc tcgccgagca ggctgcacgc cgcgaacgca tccgcgccta
     4861 tcacgacgac gagggccact cttggccgca aacggccaaa catttcgggc tgcatctgga
     4921 caccgttaag cgactcggct atcgggcgag gaaagagcgt gcggcagaac aggaagcggc
     4981 tcaaaaggcc cacaacgaag ccgacaatcc accgctgttc taacgcaatt ggggagcggg
     5041 tgtcgcgggg gttccgtggg gggttccgtt gcaacgggtc ggacaggtaa aagtcctggt
     5101 agacgctagt tttctggttt gggccatgcc tgtctcgttg cgtgtttcgt tgcgcccgtt
     5161 ttgaatacca gccagacgag acggggttct acgaatcttg gtcgatacca agccatttcc
     5221 gctgaatatc ggggagctca ccgccagaat cggtggttgt ggtgatgtac gtggcgaact
     5281 c
//
