LOCUS       sequence-348176-        5011 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of APEX2 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 5011)
  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 5011)
  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-APEX2m
FEATURES             Location/Qualifiers
     source          1..5011
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1466..1485)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1477..2292
                     /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(1550..1569)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2623..3211
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3112..3131
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      4494..4540
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gcctgagagt tgcacaggag ttgcaacccg gtagccttgt tcacgacgag aggagaccta
       61 gttggcacgt cgcggatggg gatcgctgaa gactcagcgc agcgggagga tccaagcctc
      121 atacgtcaac ccgcaggacg gtgtgaggta ctacgcgctg cagacctacg acaacaagat
      181 ggacgccgaa gcctggctcg cgggcgagaa gcggctcatc gagatggaga cctggacccc
      241 tccacaggac cgggcgaaga aggcagccgc cagcgccatc acgctggagg agtacacccg
      301 gaagtggctc gtggagcgcg acctcgcaga cggcaccagg gatctgtaca gcgggcacgc
      361 ggagcgccgc atctacccgg tgctaggtga agtggcggtc acagagatga cgccagctct
      421 ggtgcgtgcg tggtgggccg ggatgggtag gaagcacccg actgcccgcc ggcatgccta
      481 caacgtcctc cgggcggtga tgaacacagc ggtcgaggac aagctgatcg cagagaaccc
      541 gtgccggatc gagcagaagg cagccgatga gcgcgacgta gaggcgctga cgcctgagga
      601 gctggacatc gtcgccgctg agatcttcgt gcactaccgg atcgcggcat acatcctggc
      661 gtggacgagc ctccggttcg gagagctgat cgagcttcgc cgcaaggaca tcgtggacga
      721 cggcatgacg atgaagctcc gggtgcgccg tggcgcttcc cgcgtgggga acaagatcgt
      781 cgttggcaac gccaagaccg tccggtcgaa gcgtcctgtg acggttccgc ctcacgtcgc
      841 ggagatgatc cgagcgcaca tgaaggaccg tacgaagatg aacaagggcc ccgaggcatt
      901 cctggtgacc acgacgcagg gcaaccggct gtcgaagtcc gcgttcacca agtcgctgaa
      961 gcgtggctac gccaagatcg gtcggccgga actccgcatc cacgacctcc gcgctgtcgg
     1021 cgctacgttc gccgctcagg caggtgcgac gaccaaggag ctgatggccc gtctcggtca
     1081 cacgactcct aggatggcga tgaagtacca gatggcgtct gaggcccgcg acgaggctat
     1141 cgctgaggcg atgtccaagc tggccaagac ctcctgaaac gcaaaaagcc cccctcccaa
     1201 ggacactgag tcctaaagag gggggtttct tgtcagtacg cgaagaacca cgcctggccg
     1261 cgagcgccag caccgccgct ctgtgcggag acctgggcac cagccccgcc gccgccagga
     1321 gcattgccgt tcccgccagc tagccacctg acgtcggggg ggggggaaag ccacgttgtg
     1381 tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa
     1441 ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg
     1501 tcttgctcga ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     1561 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagccccat
     1621 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     1681 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     1741 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccgggaaaac agcattccag
     1801 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     1861 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt
     1921 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     1981 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataatct tttgccattc
     2041 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     2101 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     2161 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     2221 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
     2281 gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact
     2341 tgacgggacg gcggctttgt tgaataaatc gaacttttgc tgagttgaag gatcagatca
     2401 cgcatcttcc cgacaacgca gaccgttccg tggcaaagca aaagttcaaa atcaccaact
     2461 ggtccaccta caacaaagct ctcaccaacc gtggctccct cactttctgg ctggatgatg
     2521 gggcgattca ggcctggtat gagtcagcaa caccttcttc acgaggcaga cctcactagt
     2581 tccatgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2641 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     2701 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2761 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2821 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2881 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     2941 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3001 cctacagcgt gagctatgag aaagcgccac gcttcccgag gggagaaagg cggacaggta
     3061 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3121 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3181 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3241 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     3301 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     3361 gcgcaacgcg tgcggccgcg gtaccagatc tttaaatcta gaggtgacca caacgcgccc
     3421 gctttgatcg gggacgtctg cggccgacca tttacgggtc ttgttgtcgt tggcggtcat
     3481 gggccgaaca tactcacccg gatcggaggg ccgaggacaa ggtcgaacga ggggcatgac
     3541 ccggtgcggg gcttcttgca ctcggcatag gcgagtgcta agaataacgt tggcactcgc
     3601 gaccgggata cgactcacta taggtaccgg tgataccagc atcgtcttga tgcccttggc
     3661 agcaccctgc taaggaggca acaagatgca tatgggcaag agctacccga ccgtcagcgc
     3721 cgactaccag gacgccgtgg agaaggccaa gaagaagctg cgcggcttca tcgccgagaa
     3781 gcggtgcgcc ccgctgatgc tgcgcctggc gttccacagc gcgggcacct tcgacaaggg
     3841 caccaagacc ggcggcccgt tcggcaccat caagcacccg gcggagctgg cgcacagcgc
     3901 caacaacggc ctggacatcg cggtccggct gctggagccg ctgaaggcgg agttcccgat
     3961 cctgagctac gccgacttct accagctggc gggcgtcgtg gccgtcgagg tgaccggcgg
     4021 cccgaaggtg ccgttccacc cgggccggga ggacaagccg gagccgccgc cggagggccg
     4081 cctgccggac ccgaccaagg gctcggacca cctgcgggac gtcttcggca aggcgatggg
     4141 cctgaccgac caggacatcg tggccctgag cggcggccac accatcggcg cggcccacaa
     4201 ggagcgcagc ggcttcgagg gcccgtggac cagcaacccg ctgatcttcg acaactcgta
     4261 cttcaccgag ctgctgagcg gcgagaagga aggcctgctg cagctgccga gcgacaaggc
     4321 gctgctgagc gacccggtct tccggcccct ggtggacaag tacgcggccg acgaggacgc
     4381 cttcttcgcc gactacgccg aggcccacca gaagctgtcc gagctgggct tcgccgacgc
     4441 caagcttatc gatgtcgacg tagttaacta gcgtacgatc gactgccagg catcaaataa
     4501 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat gccatcatgg ccgcggtgat
     4561 cagctagagc cgtgaacgac agggcgaacg ccagcccgcc gacggcgagg gttccgaccg
     4621 ctgcaactcc cggtgcaacc ttgtcccggt ctattctctt cactgcacca gctccaatct
     4681 ggtgtgaatg cccctcgtct gttcgcgcag gcggggggct ctattcgttt gtcagcatcg
     4741 aaagtagcca gatcagggat gcgttgcaac cgcgtatgcc caggtcagaa gagtcgcaca
     4801 agagttgcag acccctggaa agaaaaatgg ccagagggcg aaaacaccct ctgaccagcg
     4861 gagcgggcga cgggaatcga acccgcgtag ctagtttgga agaatgggtg tctgccgacc
     4921 acatatatgg gccggtcaag ataggttttt accccctctc ggctgcatcc tctaagtgga
     4981 aagaaattgc aggtcgtaga agcgcgttga a
//
