LOCUS       sequence-166560-        5293 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 5293)
  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 5293)
  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: pMyBADNT-kan
FEATURES             Location/Qualifiers
     source          1..5293
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(636..1514)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1541..1825
                     /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     1748..1767
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1862..1879
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1892..1912
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      2009..2055
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2095..2910)
                     /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     2818..2837
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3249..3837
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3738..3757
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcagccgact gtccgcaaaa agcggcgcgt gaccgccgcc gaaggcgctc gaatcaccgg
       61 actatccgaa cgccacgtcg tccggctcgt ggcgcaggaa cgcagcgagt ggctcgccga
      121 gcaggctgca cgccgcgaac gcatccgcgc ctatcacgac gacgagggcc actcttggcc
      181 gcaaacggcc aaacatttcg ggctgcatct ggacaccgtt aagcgactcg gctatcgggc
      241 gaggaaagag cgtgcggcag aacaggaagc ggctcaaaag gcccacaacg aagccgacaa
      301 tccaccgctg ttctaacgca attggggagc gggtgtcgcg ggggttccgt ggggggttcc
      361 gttgcaacgg gtcggacagg taaaagtcct ggtagacgct agttttctgg tttgggccat
      421 gcctgtctcg ttgcgtgttt cgttgcgccc gttttgaata ccagccagac gagacggggt
      481 tctacgaatc ttggtcgata ccaagccatt tccgctgaat atcggggagc tcaccgccag
      541 aatcggtggt tgtggtgatg tacgtggcga actccgttgt agtgcctgtg gtggcatccg
      601 tggcgcggcc gcggtaccag atctttaaat ctagattatg acaacttgac ggctacatca
      661 ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta
      721 aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata
      781 ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag
      841 cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag
      901 caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg
      961 tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct
     1021 tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc
     1081 ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc
     1141 gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca
     1201 tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga
     1261 tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa
     1321 acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata
     1381 taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc
     1441 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt
     1501 tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat
     1561 tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta
     1621 accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt
     1681 aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca
     1741 ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta
     1801 tcgcaactct ctactgtttc tccatacccg ttttttgggc taacaggagg aattaaccat
     1861 gcaccaccac caccaccacc cgtcggccgg cgagaacctg tacttccagg gcgccatgga
     1921 cgataaggta ccggatccga attcgatatc gctagcaagc ttatcgatgt cgacgtagtt
     1981 aactagcgta cgatcgactg ccaggcatca aataaaacga aaggctcagt cgaaagactg
     2041 ggcctttcgt tttatgccat catggccgcg gtgatcagct agagggggcg ctgattagaa
     2101 aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata
     2161 tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat
     2221 ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa
     2281 tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc
     2341 cggtgagaat ggcaaaagat tatgcatttc tttccagact tgttcaacag gccagccatt
     2401 acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg
     2461 agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa
     2521 ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc
     2581 taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg
     2641 agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct
     2701 gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc
     2761 tggcgcatgg ggcttcccat acaagcgata gattgtcgca cctgattgcc cgacattatc
     2821 gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctcga
     2881 gcaagacgtt tcccgttgaa tatggctcat agcggacctc tattcacagg gtacgggccg
     2941 gcttaattcc gcacggccgg tcgcgacacg gcctgtccgc accgcggatc aggcgttgac
     3001 gatgacgggc tggtcggcca cgtcggggag tcaggcaact atggatgaac gaaatagaca
     3061 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     3121 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     3181 cctttttgat aatctcatgc tctagttcca tgagcgtcag accccgtaga aaagatcaaa
     3241 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3301 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3361 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     3421 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3481 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3541 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3601 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     3661 cccgagggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3721 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     3781 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     3841 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     3901 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     3961 accgctcgcc gcagccgaac gaccgagcgc aacgcgtgcg gccgcacgcg tgagcccacc
     4021 agctccgtaa gttcgggcgc tgtgtggctc gtacccgcgc attcaggcgg cagggggtct
     4081 aacgggtcta aggcggcgtg tacggccgcc acagcggctc tcagcggccc ggaaacgtcc
     4141 tcgaaacgac gcatgtgttc ctcctggttg gtacaggtgg ttgggggtgc tcggctgtcg
     4201 ctggtgttcc accaccaggg ctcgacggga gagcggggga gtgtgcagtt gtggggtggc
     4261 ccctcagcga aatatctgac ttggagctcg tgtcggacca tacaccggtg attaatcgtg
     4321 gtctactacc aagcgtgagc cacgtcgccg acgaatttga gcagctctgg ctgccgtact
     4381 ggccgctggc aagcgacgat ctgctcgagg ggatctaccg ccaaagccgc gcgtcggccc
     4441 taggccgccg gtacatcgag gcgaacccaa cagcgctggc aaacctgctg gtcgtggacg
     4501 tagaccatcc agacgcagcg ctccgagcgc tcagcgcccg ggggtcccat ccgctgccca
     4561 acgcgatcgt gggcaatcgc gccaacggcc acgcacacgc agtgtgggca ctcaacgccc
     4621 ctgttccacg caccgaatac gcgcggcgta agccgctcgc atacatggcg gcgtgcgccg
     4681 aaggccttcg gcgcgccgtc gatggcgacc gcagttactc aggcctcatg accaaaaacc
     4741 ccggccacat cgcctgggaa acggaatggc tccactcaga tctctacaca ctcagccaca
     4801 tcgaggccga gctcggcgcg aacatgccac cgccgcgctg gcgtcagcag accacgtaca
     4861 aagcggctcc gacgccgcta gggcggaatt gcgcactgtt cgattccgtc aggttgtggg
     4921 cctatcgtcc cgccctcatg cggatctacc tgccgacccg gaacgtggac ggactcggcc
     4981 gcgcgatcta tgccgagtgc cacgcgcgaa acgccgaatt tccgtgcaac gacgtgtgtc
     5041 ccggaccgct accggacagc gaggtccgcg ccatcgccaa cagcatttgg cgttggatca
     5101 caaccaagtc gcgcatttgg gcggacggga tcgtggtcta cgaggccaca ctcagtgcgc
     5161 gccagtcggc catctcgcgg aagggcgcag cagcgcgcac ggcggcgagc acagttgcgc
     5221 ggcgcgcaaa gtccgcgtca gccatgcatg gaggcattgc tatgagcgac ggctacagcg
     5281 acggctacaa ccg
//
