LOCUS       sequence-169520-        5409 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression in mycobacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5409)
  AUTHORS   Beckham KSH, Staack S, Wilmanns M, Parret AHA
  TITLE     The pMy vector series: A versatile cloning platform for the 
            recombinant production of mycobacterial proteins in Mycobacterium 
            smegmatis.
  JOURNAL   Protein Sci. 2020 Oct 2. doi: 10.1002/pro.3962.
  PUBMED    33006405
REFERENCE   2  (bases 1 to 5409)
  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: "Protein 
            Sci. 2020 Oct 2. doi: 10.1002/pro.3962."
COMMENT     Sequence Label: pMyBADC-kan
FEATURES             Location/Qualifiers
     source          1..5409
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      14..602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     503..522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2717..3595)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3622..3906
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     3829..3848
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3993..4010
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      4069..4115
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(4155..4970)
                     /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     4878..4897
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4962..4981
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
       61 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
      121 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
      181 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
      241 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
      301 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
      361 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
      421 cgcttcccga ggggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
      481 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
      541 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
      601 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      661 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
      721 ctgataccgc tcgccgcagc cgaacgaccg agcgcaacgc gtgcggccgc acgcgtgagc
      781 ccaccagctc cgtaagttcg ggcgctgtgt ggctcgtacc cgcgcattca ggcggcaggg
      841 ggtctaacgg gtctaaggcg gcgtgtacgg ccgccacagc ggctctcagc ggcccggaaa
      901 cgtcctcgaa acgacgcatg tgttcctcct ggttggtaca ggtggttggg ggtgctcggc
      961 tgtcgctggt gttccaccac cagggctcga cgggagagcg ggggagtgtg cagttgtggg
     1021 gtggcccctc agcgaaatat ctgacttgga gctcgtgtcg gaccatacac cggtgattaa
     1081 tcgtggtcta ctaccaagcg tgagccacgt cgccgacgaa tttgagcagc tctggctgcc
     1141 gtactggccg ctggcaagcg acgatctgct cgaggggatc taccgccaaa gccgcgcgtc
     1201 ggccctaggc cgccggtaca tcgaggcgaa cccaacagcg ctggcaaacc tgctggtcgt
     1261 ggacgtagac catccagacg cagcgctccg agcgctcagc gcccgggggt cccatccgct
     1321 gcccaacgcg atcgtgggca atcgcgccaa cggccacgca cacgcagtgt gggcactcaa
     1381 cgcccctgtt ccacgcaccg aatacgcgcg gcgtaagccg ctcgcataca tggcggcgtg
     1441 cgccgaaggc cttcggcgcg ccgtcgatgg cgaccgcagt tactcaggcc tcatgaccaa
     1501 aaaccccggc cacatcgcct gggaaacgga atggctccac tcagatctct acacactcag
     1561 ccacatcgag gccgagctcg gcgcgaacat gccaccgccg cgctggcgtc agcagaccac
     1621 gtacaaagcg gctccgacgc cgctagggcg gaattgcgca ctgttcgatt ccgtcaggtt
     1681 gtgggcctat cgtcccgccc tcatgcggat ctacctgccg acccggaacg tggacggact
     1741 cggccgcgcg atctatgccg agtgccacgc gcgaaacgcc gaatttccgt gcaacgacgt
     1801 gtgtcccgga ccgctaccgg acagcgaggt ccgcgccatc gccaacagca tttggcgttg
     1861 gatcacaacc aagtcgcgca tttgggcgga cgggatcgtg gtctacgagg ccacactcag
     1921 tgcgcgccag tcggccatct cgcggaaggg cgcagcagcg cgcacggcgg cgagcacagt
     1981 tgcgcggcgc gcaaagtccg cgtcagccat gcatggaggc attgctatga gcgacggcta
     2041 cagcgacggc tacaaccggc agccgactgt ccgcaaaaag cggcgcgtga ccgccgccga
     2101 aggcgctcga atcaccggac tatccgaacg ccacgtcgtc cggctcgtgg cgcaggaacg
     2161 cagcgagtgg ctcgccgagc aggctgcacg ccgcgaacgc atccgcgcct atcacgacga
     2221 cgagggccac tcttggccgc aaacggccaa acatttcggg ctgcatctgg acaccgttaa
     2281 gcgactcggc tatcgggcga ggaaagagcg tgcggcagaa caggaagcgg ctcaaaaggc
     2341 ccacaacgaa gccgacaatc caccgctgtt ctaacgcaat tggggagcgg gtgtcgcggg
     2401 ggttccgtgg ggggttccgt tgcaacgggt cggacaggta aaagtcctgg tagacgctag
     2461 ttttctggtt tgggccatgc ctgtctcgtt gcgtgtttcg ttgcgcccgt tttgaatacc
     2521 agccagacga gacggggttc tacgaatctt ggtcgatacc aagccatttc cgctgaatat
     2581 cggggagctc accgccagaa tcggtggttg tggtgatgta cgtggcgaac tccgttgtag
     2641 tgcctgtggt ggcatccgtg gcgcggccgc ggtaccagat ctttaaatct agataaagaa
     2701 gtgacgcggt cctagattat gacaacttga cggctacatc attcactttt tcttcacaac
     2761 cggcacggaa ctcgctcggg ctggccccgg tgcatttttt aaatacccgc gagaaataga
     2821 gttgatcgtc aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg gtggtgctca
     2881 aaagcagctt cgcctggctg atacgttggt cctcgcgcca gcttaagacg ctaatcccta
     2941 actgctggcg gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc
     3001 tggcgatatc aaaattgctg tctgccaggt gatcgctgat gtactgacaa gcctcgcgta
     3061 cccgattatc catcggtgga tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca
     3121 attgctcaag cagatttatc gccagcagct ccgaatagcg cccttcccct tgcccggcgt
     3181 taatgatttg cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga
     3241 accccgtatt ggcaaatatt gacggccagt taagccattc atgccagtag gcgcgcggac
     3301 gaaagtaaac ccactggtga taccattcgc gagcctccgg atgacgaccg tagtgatgaa
     3361 tctctcctgg cgggaacagc aaaatatcac ccggtcggca aacaaattct cgtccctgat
     3421 ttttcaccac cccctgaccg cgaatggtga gattgagaat ataacctttc attcccagcg
     3481 gtcggtcgat aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa cccgccacca
     3541 gatgggcatt aaacgagtat cccggcagca ggggatcatt ttgcgcttca gccatacttt
     3601 tcatactccc gccattcaga gaagaaacca attgtccata ttgcatcaga cattgccgtc
     3661 actgcgtctt ttactggctc ttctcgctaa ccaaaccggt aaccccgctt attaaaagca
     3721 ttctgtaaca aagcgggacc aaagccatga caaaaacgcg taacaaaagt gtctataatc
     3781 acggcagaaa agtccacatt gattatttgc acggcgtcac actttgctat gccatagcat
     3841 ttttatccat aagattagcg gatcctacct gacgcttttt atcgcaactc tctactgttt
     3901 ctccataccc gttttttggg ctaacaggag gaattaaccc catgggctcg gccggtaccg
     3961 gatccgaatt cgatatcgct agccaagctt cgcaccacca ccaccaccac tgataacagc
     4021 ttatcgatgt cgacgtagtt aactagcgta cgatcgactg ccaggcatca aataaaacga
     4081 aaggctcagt cgaaagactg ggcctttcgt tttatgccat catggccgcg gtgatcagct
     4141 agagggggcg ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata
     4201 tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca
     4261 ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca
     4321 acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca
     4381 ccatgagtga cgactgaatc cggtgagaat ggcaaaagat tatgcatttc tttccagact
     4441 tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta
     4501 ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta
     4561 caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca
     4621 cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg
     4681 agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat
     4741 tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg
     4801 ccatgtttca gaaacaactc tggcgcatgg ggcttcccat acaagcgata gattgtcgca
     4861 cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg
     4921 gaatttaatc gcggcctcga gcaagacgtt tcccgttgaa tatggctcat aacacccctt
     4981 gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt
     5041 gcaatgtaac atcagagatt ttgagacaca acgtcgcttt gttgagcgga cctctattca
     5101 cagggtacgg gccggcttaa ttccgcacgg ccggtcgcga cacggcctgt ccgcaccgcg
     5161 gatcaggcgt tgacgatgac gggctggtcg gccacgtcgg ggagtcaggc aactatggat
     5221 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5281 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5341 atctaggtga agatcctttt tgataatctc atgctctagt tccatgagcg tcagaccccg
     5401 tagaaaaga
//
