LOCUS       sequence-348178-        5106 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of APEX2 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 5106)
  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 5106)
  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-APEX2m
FEATURES             Location/Qualifiers
     source          1..5106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1157..1203
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1353..1372)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1364..2179
                     /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(1437..1456)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2510..3098
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2999..3018
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 actccgttgt agtgcctgtg gtggcatccg tggcgcggcc gcggtaccag atctttaaat
       61 ctagaggtga ccacaacgcg cccgctttga tcggggacgt ctgcggccga ccatttacgg
      121 gtcttgttgt cgttggcggt catgggccga acatactcac ccggatcgga gggccgagga
      181 caaggtcgaa cgaggggcat gacccggtgc ggggcttctt gcactcggca taggcgagtg
      241 ctaagaataa cgttggcact cgcgaccggg atacgactca ctataggtac cggtgatacc
      301 agcatcgtct tgatgccctt ggcagcaccc tgctaaggag gcaacaagat gcatatgggc
      361 aagagctacc cgaccgtcag cgccgactac caggacgccg tggagaaggc caagaagaag
      421 ctgcgcggct tcatcgccga gaagcggtgc gccccgctga tgctgcgcct ggcgttccac
      481 agcgcgggca ccttcgacaa gggcaccaag accggcggcc cgttcggcac catcaagcac
      541 ccggcggagc tggcgcacag cgccaacaac ggcctggaca tcgcggtccg gctgctggag
      601 ccgctgaagg cggagttccc gatcctgagc tacgccgact tctaccagct ggcgggcgtc
      661 gtggccgtcg aggtgaccgg cggcccgaag gtgccgttcc acccgggccg ggaggacaag
      721 ccggagccgc cgccggaggg ccgcctgccg gacccgacca agggctcgga ccacctgcgg
      781 gacgtcttcg gcaaggcgat gggcctgacc gaccaggaca tcgtggccct gagcggcggc
      841 cacaccatcg gcgcggccca caaggagcgc agcggcttcg agggcccgtg gaccagcaac
      901 ccgctgatct tcgacaactc gtacttcacc gagctgctga gcggcgagaa ggaaggcctg
      961 ctgcagctgc cgagcgacaa ggcgctgctg agcgacccgg tcttccggcc cctggtggac
     1021 aagtacgcgg ccgacgagga cgccttcttc gccgactacg ccgaggccca ccagaagctg
     1081 tccgagctgg gcttcgccga cgccaagctt atcgatgtcg acgtagttaa ctagcgtacg
     1141 atcgactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     1201 tatgccatca tggccgcggt gatcagctag ccacctgacg tcgggggggg gggaaagcca
     1261 cgttgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
     1321 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     1381 ggaaacgtct tgctcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     1441 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     1501 gccccatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     1561 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     1621 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg ggaaaacagc
     1681 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     1741 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     1801 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     1861 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataatctttt
     1921 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     1981 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     2041 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     2101 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     2161 gctcgatgag tttttctaat cagaattggt taattggttg taacactggc agagcattac
     2221 gctgacttga cgggacggcg gctttgttga ataaatcgaa cttttgctga gttgaaggat
     2281 cagatcacgc atcttcccga caacgcagac cgttccgtgg caaagcaaaa gttcaaaatc
     2341 accaactggt ccacctacaa caaagctctc accaaccgtg gctccctcac tttctggctg
     2401 gatgatgggg cgattcaggc ctggtatgag tcagcaacac cttcttcacg aggcagacct
     2461 cactagttcc atgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     2521 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     2581 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     2641 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2701 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2761 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2821 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2881 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgagggg agaaaggcgg
     2941 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3001 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     3061 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     3121 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     3181 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     3241 cgaccgagcg caacgcgtga gcccaccagc tccgtaagtt cgggcgctgt gtggctcgta
     3301 cccgcgcatt caggcggcag ggggtctaac gggtctaagg cggcgtgtac ggccgccaca
     3361 gcggctctca gcggcccgga aacgtcctcg aaacgacgca tgtgttcctc ctggttggta
     3421 caggtggttg ggggtgctcg gctgtcgctg gtgttccacc accagggctc gacgggagag
     3481 cgggggagtg tgcagttgtg gggtggcccc tcagcgaaat atctgacttg gagctcgtgt
     3541 cggaccatac accggtgatt aatcgtggtc tactaccaag cgtgagccac gtcgccgacg
     3601 aatttgagca gctctggctg ccgtactggc cgctggcaag cgacgatctg ctcgagggga
     3661 tctaccgcca aagccgcgcg tcggccctag gccgccggta catcgaggcg aacccaacag
     3721 cgctggcaaa cctgctggtc gtggacgtag accatccaga cgcagcgctc cgagcgctca
     3781 gcgcccgggg gtcccatccg ctgcccaacg cgatcgtggg caatcgcgcc aacggccacg
     3841 cacacgcagt gtgggcactc aacgcccctg ttccacgcac cgaatacgcg cggcgtaagc
     3901 cgctcgcata catggcggcg tgcgccgaag gccttcggcg cgccgtcgat ggcgaccgca
     3961 gttactcagg cctcatgacc aaaaaccccg gccacatcgc ctgggaaacg gaatggctcc
     4021 actcagatct ctacacactc agccacatcg aggccgagct cggcgcgaac atgccaccgc
     4081 cgcgctggcg tcagcagacc acgtacaaag cggctccgac gccgctaggg cggaattgcg
     4141 cactgttcga ttccgtcagg ttgtgggcct atcgtcccgc cctcatgcgg atctacctgc
     4201 cgacccggaa cgtggacgga ctcggccgcg cgatctatgc cgagtgccac gcgcgaaacg
     4261 ccgaatttcc gtgcaacgac gtgtgtcccg gaccgctacc ggacagcgag gtccgcgcca
     4321 tcgccaacag catttggcgt tggatcacaa ccaagtcgcg catttgggcg gacgggatcg
     4381 tggtctacga ggccacactc agtgcgcgcc agtcggccat ctcgcggaag ggcgcagcag
     4441 cgcgcacggc ggcgagcaca gttgcgcggc gcgcaaagtc cgcgtcagcc atggaggcat
     4501 tgctatgagc gacggctaca gcgacggcta caaccggcag ccgactgtcc gcaaaaagcg
     4561 gcgcgtgacc gccgccgaag gcgctcgaat caccggacta tccgaacgcc acgtcgtccg
     4621 gctcgtggcg caggaacgca gcgagtggct cgccgagcag gctgcacgcc gcgaacgcat
     4681 ccgcgcctat cacgacgacg agggccactc ttggccgcaa acggccaaac atttcgggct
     4741 gcatctggac accgttaagc gactcggcta tcgggcgagg aaagagcgtg cggcagaaca
     4801 ggaagcggct caaaaggccc acaacgaagc cgacaatcca ccgctgttct aacgcaattg
     4861 gggagcgggt gtcgcggggg ttccgtgggg ggttccgttg caacgggtcg gacaggtaaa
     4921 agtcctggta gacgctagtt ttctggtttg ggccatgcct gtctcgttgc gtgtttcgtt
     4981 gcgcccgttt tgaataccag ccagacgaga cggggttcta cgaatcttgg tcgataccaa
     5041 gccatttccg ctgaatatcg gggagctcac cgccagaatc ggtggttgtg gtgatgtacg
     5101 tggcga
//
