LOCUS       sequence-169523-        6837 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 6837)
  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 6837)
  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: pMyNT-kan
FEATURES             Location/Qualifiers
     source          1..6837
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      27..615
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     516..535
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             5363..5380
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5393..5413
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      5510..5556
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(5596..6411)
                     /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     6319..6338
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6403..6422
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
       61 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
      121 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
      181 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
      241 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
      301 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
      361 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
      421 tgagaaagcg ccacgcttcc cgaggggaga aaggcggaca ggtatccggt aagcggcagg
      481 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
      541 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
      601 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
      661 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
      721 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcaa cgcgtgcggc
      781 cgcacgcgtg agcccaccag ctccgtaagt tcgggcgctg tgtggctcgt acccgcgcat
      841 tcaggcggca gggggtctaa cgggtctaag gcggcgtgta cggccgccac agcggctctc
      901 agcggcccgg aaacgtcctc gaaacgacgc atgtgttcct cctggttggt acaggtggtt
      961 gggggtgctc ggctgtcgct ggtgttccac caccagggct cgacgggaga gcgggggagt
     1021 gtgcagttgt ggggtggccc ctcagcgaaa tatctgactt ggagctcgtg tcggaccata
     1081 caccggtgat taatcgtggt ctactaccaa gcgtgagcca cgtcgccgac gaatttgagc
     1141 agctctggct gccgtactgg ccgctggcaa gcgacgatct gctcgagggg atctaccgcc
     1201 aaagccgcgc gtcggcccta ggccgccggt acatcgaggc gaacccaaca gcgctggcaa
     1261 acctgctggt cgtggacgta gaccatccag acgcagcgct ccgagcgctc agcgcccggg
     1321 ggtcccatcc gctgcccaac gcgatcgtgg gcaatcgcgc caacggccac gcacacgcag
     1381 tgtgggcact caacgcccct gttccacgca ccgaatacgc gcggcgtaag ccgctcgcat
     1441 acatggcggc gtgcgccgaa ggccttcggc gcgccgtcga tggcgaccgc agttactcag
     1501 gcctcatgac caaaaacccc ggccacatcg cctgggaaac ggaatggctc cactcagatc
     1561 tctacacact cagccacatc gaggccgagc tcggcgcgaa catgccaccg ccgcgctggc
     1621 gtcagcagac cacgtacaaa gcggctccga cgccgctagg gcggaattgc gcactgttcg
     1681 attccgtcag gttgtgggcc tatcgtcccg ccctcatgcg gatctacctg ccgacccgga
     1741 acgtggacgg actcggccgc gcgatctatg ccgagtgcca cgcgcgaaac gccgaatttc
     1801 cgtgcaacga cgtgtgtccc ggaccgctac cggacagcga ggtccgcgcc atcgccaaca
     1861 gcatttggcg ttggatcaca accaagtcgc gcatttgggc ggacgggatc gtggtctacg
     1921 aggccacact cagtgcgcgc cagtcggcca tctcgcggaa gggcgcagca gcgcgcacgg
     1981 cggcgagcac agttgcgcgg cgcgcaaagt ccgcgtcagc catgcatgga ggcattgcta
     2041 tgagcgacgg ctacagcgac ggctacaacc ggcagccgac tgtccgcaaa aagcggcgcg
     2101 tgaccgccgc cgaaggcgct cgaatcaccg gactatccga acgccacgtc gtccggctcg
     2161 tggcgcagga acgcagcgag tggctcgccg agcaggctgc acgccgcgaa cgcatccgcg
     2221 cctatcacga cgacgagggc cactcttggc cgcaaacggc caaacatttc gggctgcatc
     2281 tggacaccgt taagcgactc ggctatcggg cgaggaaaga gcgtgcggca gaacaggaag
     2341 cggctcaaaa ggcccacaac gaagccgaca atccaccgct gttctaacgc aattggggag
     2401 cgggtgtcgc gggggttccg tggggggttc cgttgcaacg ggtcggacag gtaaaagtcc
     2461 tggtagacgc tagttttctg gtttgggcca tgcctgtctc gttgcgtgtt tcgttgcgcc
     2521 cgttttgaat accagccaga cgagacgggg ttctacgaat cttggtcgat accaagccat
     2581 ttccgctgaa tatcggggag ctcaccgcca gaatcggtgg ttgtggtgat gtacgtggcg
     2641 aactccgttg tagtgcctgt ggtggcatcc gtggcgcggc cgcggtacca gatctttaaa
     2701 tctagataaa gaagtgacgc ggtctcaagc gtcgagcgtc gccagcgtgt cgaggatgtc
     2761 gaagtcgtag ccgtcggcgc tggcgatgta gacctgctgg tcgaattgac tgtcgcgcat
     2821 acacatcggg ccccggggcc cgtcgaaccc gacatcgtgc gcggatgcca tcaggtccgg
     2881 tatctcgggg gagtgggccc gctggaagat ggcctcgagc gcaagcagac cctcgtaaca
     2941 ggattcggcc atcgcgttga gcggtggcgc gtcggcgccg tagcgggcga cgtagctgcc
     3001 catcaggtcc atcgcacccg cggtggccag tgaactgaag tacgccgcgg cgacatagag
     3061 gttttcggtg gagccggcgc cgctggccag cagcatgttc tcctccatca gcgggctgaa
     3121 ccgcgccatg cggtcgtgcc cgccggcgcg cgcgaactcg cggttgaaca acacggcgtc
     3181 ctggccgacg agcagcatca acacggcctg cgcccccgac gcgatggcct tgcggacagg
     3241 tgcgcggaaa tcgtcggtgc cgtacgggac gtagatctcc cgtctgagct cgaggtccag
     3301 atctcggcag tacgcgcggg cggccgcggc ggaacggcgc ggccagatgt agtcatcgcc
     3361 gaccaggcac caggaccgga tgccgaagtg gtcgcgcagc caggcgagcg cgggcgcgat
     3421 ctggatctgc ggtgtctcgc ctgtgcagaa cacgcccggt gtgcgttcac cgccctcgta
     3481 caacgaggtg tagacgtacg ggatgcggtc gcggaccacc ggggagatgc ggttgcgcac
     3541 ggccgagatg tgccagccgg tcacggcgtc gagaccgtga cctcgcaacc ggtcggcgac
     3601 ggtccgggcg acgtcgtcgc cgggcgctcc gccgtcgagc acctcgatgg tgaccttgcg
     3661 gccctgcagg ccgcctcggt cgttgacctc cttggccgcg agctcggcca cggcctcgca
     3721 cgaaggcgcg aagattcccg ctggcccttg aagcggaatc accagcccga cgcggaactc
     3781 aacctcgccg tcctgcactc cagatcaccg tcgatcccgt gtagtctgcg cttcaaaggt
     3841 ttctagcaga aataattcat tctgaacaga ccccgccgtc gacacgagga gacccaacat
     3901 ggccgccgga cagcagcgcc gccccaacct cctgctgccg ttggtgcgtc tgacccacct
     3961 cgcggagtcg gcgatcgaac gcgtgctcgc ggactcgtcg ctcaagatcg aggactggcg
     4021 cgtgctcgac gagttggccg gacggcgcac cgtgcccatg agcgatctcg cgcaggccac
     4081 gctgatcacg ggtccgactc tcaccagaac cgtcgatcgc cttgtgtcgc aagggatcat
     4141 ctaccggact gccgatctgc atgaccgccg gcgggtgctc gtggcgttga ccccgcgggg
     4201 gcggacgctg cgcaaccgcc tggtggacgc ggtagccgag gccgagtgtg cggcttttga
     4261 atcgtgcggg ctggacgtcg accagttgcg cgaactcgtc gacaccacct cgaatttgac
     4321 ttcgtaacca cccgcgcccg gcgcgggcgt tcacccttga cttttatttt catctggata
     4381 tatttcgggt gaatggaaag gggtgaccat gccgacctac acattccgtt gttcccactg
     4441 cggtcccttc gatctcacct gcgcgatctc cgagcgcgat gcggcggcga cctgtccgga
     4501 gtgccggacg ccggcgcgcc gggtcttcgg ttcggtaggg ctgacgacat tcaccgcggg
     4561 acatcaccgc gcattcgacg cggcgtccgc gagcgccgaa agtcccacgg tggtgaagtc
     4621 gattcccgca ggcgcggacc gcccgcgggc cccgcgccgc aatcccggtc taccgagtct
     4681 gccgaggtac tagcgacatg ggtggcgtcg ggctcttcta cgtgggtgcg gtgctcatca
     4741 tcgacgggct gatgctgctg ggccgcatca gcccacgagg cgcaacaccg ctgaacttct
     4801 tcgtcggcgg actgcaggtg gtgacgccta cggtgctgat cctgcagtcc ggcggagacg
     4861 cggccgtgat cttcgcggcc tccgggctct acctgttcgg cttcacctac ctgtgggtgg
     4921 ccatcaacaa cgtgaccgac tgggacggag aaggtctcgg atggttctcg ctgttcgtcg
     4981 cgatcgccgc actcggctac tcgtggcacg cgttcaccgc cgaggccgac ccggcgttcg
     5041 gggtgatctg gctgctgtgg gcagtgctgt ggttcatgct gttcctgctg ctcggcctgg
     5101 ggcacgacgc actggggccc gccgtcgggt tcgtcgcggt ggccgaaggc gtgatcaccg
     5161 ccgccgtgcc ggccttcctg atcgtgtcgg gcaactggga aaccggcccg ctccccgccg
     5221 cggtcatcgc cgtgatcggt tttgccgcag ttgttctcgc ataccccatc gggcgccgtc
     5281 tcgcagcgcc gtcagtcacc aaccctccac cggccgcgct cgcggccacc acccgataag
     5341 agaaagggag tccaccatga agcaccacca ccaccaccac ccgtcggccg gcgagaacct
     5401 gtacttccag ggcgccatgg acgataaggt accggatccg aattcgatat cgctagcaag
     5461 cttatcgatg tcgacgtagt taactagcgt acgatcgact gccaggcatc aaataaaacg
     5521 aaaggctcag tcgaaagact gggcctttcg ttttatgcca tcatggccgc ggtgatcagc
     5581 tagagggggc gctgattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
     5641 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     5701 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     5761 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     5821 accatgagtg acgactgaat ccggtgagaa tggcaaaaga ttatgcattt ctttccagac
     5881 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     5941 attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt
     6001 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     6061 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
     6121 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     6181 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     6241 gccatgtttc agaaacaact ctggcgcatg gggcttccca tacaagcgat agattgtcgc
     6301 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     6361 ggaatttaat cgcggcctcg agcaagacgt ttcccgttga atatggctca taacacccct
     6421 tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg
     6481 tgcaatgtaa catcagagat tttgagacac aacgtcgctt tgttgagcgg acctctattc
     6541 acagggtacg ggccggctta attccgcacg gccggtcgcg acacggcctg tccgcaccgc
     6601 ggatcaggcg ttgacgatga cgggctggtc ggccacgtcg gggagtcagg caactatgga
     6661 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     6721 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     6781 gatctaggtg aagatccttt ttgataatct catgctctag ttccatgagc gtcagac
//
