LOCUS      pMyC-kan       6811 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Protein expression in mycobacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6811)
  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 6811)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            //gt12365.cn
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Protein 
            Sci. 2020 Oct 2. doi: 10.1002/pro.3962."
COMMENT     Sequence Label: pMyC-kan
FEATURES             Location/Qualifiers
     source          1..6811
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             728..745
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      804..850
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(890..1705)
                     /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     1613..1632
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1697..1716
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2158..2746
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2647..2666
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccgaggtac tagcgacatg ggtggcgtcg ggctcttcta cgtgggtgcg gtgctcatca
       61 tcgacgggct gatgctgctg ggccgcatca gcccacgagg cgcaacaccg ctgaacttct
      121 tcgtcggcgg actgcaggtg gtgacgccta cggtgctgat cctgcagtcc ggcggagacg
      181 cggccgtgat cttcgcggcc tccgggctct acctgttcgg cttcacctac ctgtgggtgg
      241 ccatcaacaa cgtgaccgac tgggacggag aaggtctcgg atggttctcg ctgttcgtcg
      301 cgatcgccgc actcggctac tcgtggcacg cgttcaccgc cgaggccgac ccggcgttcg
      361 gggtgatctg gctgctgtgg gcagtgctgt ggttcatgct gttcctgctg ctcggcctgg
      421 ggcacgacgc actggggccc gccgtcgggt tcgtcgcggt ggccgaaggc gtgatcaccg
      481 ccgccgtgcc ggccttcctg atcgtgtcgg gcaactggga aaccggcccg ctccccgccg
      541 cggtcatcgc cgtgatcggt tttgccgcag ttgttctcgc ataccccatc gggcgccgtc
      601 tcgcagcgcc gtcagtcacc aaccctccac cggccgcgct cgcggccacc acccgataag
      661 agaaagggag tccaccatgg gctcggccgg taccggatcc gaattcgata tcgctagcca
      721 agcttcgcac caccaccacc accactgata acagcttatc gatgtcgacg tagttaacta
      781 gcgtacgatc gactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
      841 ttcgttttat gccatcatgg ccgcggtgat cagctagagg gggcgctgat tagaaaaact
      901 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
      961 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
     1021 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
     1081 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
     1141 agaatggcaa aagattatgc atttctttcc agacttgttc aacaggccag ccattacgct
     1201 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
     1261 gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     1321 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     1381 cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     1441 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
     1501 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     1561 catggggctt cccatacaag cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
     1621 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctcgagcaag
     1681 acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca
     1741 gttttattgt tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag
     1801 acacaacgtc gctttgttga gcggacctct attcacaggg tacgggccgg cttaattccg
     1861 cacggccggt cgcgacacgg cctgtccgca ccgcggatca ggcgttgacg atgacgggct
     1921 ggtcggccac gtcggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     1981 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     2041 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     2101 atctcatgct ctagttccat gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2161 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2221 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2281 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     2341 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2401 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2461 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     2521 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaggggag
     2581 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2641 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2701 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2761 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2821 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2881 cagccgaacg accgagcgca acgcgtgcgg ccgcacgcgt gagcccacca gctccgtaag
     2941 ttcgggcgct gtgtggctcg tacccgcgca ttcaggcggc agggggtcta acgggtctaa
     3001 ggcggcgtgt acggccgcca cagcggctct cagcggcccg gaaacgtcct cgaaacgacg
     3061 catgtgttcc tcctggttgg tacaggtggt tgggggtgct cggctgtcgc tggtgttcca
     3121 ccaccagggc tcgacgggag agcgggggag tgtgcagttg tggggtggcc cctcagcgaa
     3181 atatctgact tggagctcgt gtcggaccat acaccggtga ttaatcgtgg tctactacca
     3241 agcgtgagcc acgtcgccga cgaatttgag cagctctggc tgccgtactg gccgctggca
     3301 agcgacgatc tgctcgaggg gatctaccgc caaagccgcg cgtcggccct aggccgccgg
     3361 tacatcgagg cgaacccaac agcgctggca aacctgctgg tcgtggacgt agaccatcca
     3421 gacgcagcgc tccgagcgct cagcgcccgg gggtcccatc cgctgcccaa cgcgatcgtg
     3481 ggcaatcgcg ccaacggcca cgcacacgca gtgtgggcac tcaacgcccc tgttccacgc
     3541 accgaatacg cgcggcgtaa gccgctcgca tacatggcgg cgtgcgccga aggccttcgg
     3601 cgcgccgtcg atggcgaccg cagttactca ggcctcatga ccaaaaaccc cggccacatc
     3661 gcctgggaaa cggaatggct ccactcagat ctctacacac tcagccacat cgaggccgag
     3721 ctcggcgcga acatgccacc gccgcgctgg cgtcagcaga ccacgtacaa agcggctccg
     3781 acgccgctag ggcggaattg cgcactgttc gattccgtca ggttgtgggc ctatcgtccc
     3841 gccctcatgc ggatctacct gccgacccgg aacgtggacg gactcggccg cgcgatctat
     3901 gccgagtgcc acgcgcgaaa cgccgaattt ccgtgcaacg acgtgtgtcc cggaccgcta
     3961 ccggacagcg aggtccgcgc catcgccaac agcatttggc gttggatcac aaccaagtcg
     4021 cgcatttggg cggacgggat cgtggtctac gaggccacac tcagtgcgcg ccagtcggcc
     4081 atctcgcgga agggcgcagc agcgcgcacg gcggcgagca cagttgcgcg gcgcgcaaag
     4141 tccgcgtcag ccatgcatgg aggcattgct atgagcgacg gctacagcga cggctacaac
     4201 cggcagccga ctgtccgcaa aaagcggcgc gtgaccgccg ccgaaggcgc tcgaatcacc
     4261 ggactatccg aacgccacgt cgtccggctc gtggcgcagg aacgcagcga gtggctcgcc
     4321 gagcaggctg cacgccgcga acgcatccgc gcctatcacg acgacgaggg ccactcttgg
     4381 ccgcaaacgg ccaaacattt cgggctgcat ctggacaccg ttaagcgact cggctatcgg
     4441 gcgaggaaag agcgtgcggc agaacaggaa gcggctcaaa aggcccacaa cgaagccgac
     4501 aatccaccgc tgttctaacg caattgggga gcgggtgtcg cgggggttcc gtggggggtt
     4561 ccgttgcaac gggtcggaca ggtaaaagtc ctggtagacg ctagttttct ggtttgggcc
     4621 atgcctgtct cgttgcgtgt ttcgttgcgc ccgttttgaa taccagccag acgagacggg
     4681 gttctacgaa tcttggtcga taccaagcca tttccgctga atatcgggga gctcaccgcc
     4741 agaatcggtg gttgtggtga tgtacgtggc gaactccgtt gtagtgcctg tggtggcatc
     4801 cgtggcgcgg ccgcggtacc agatctttaa atctagataa agaagtgacg cggtctcaag
     4861 cgtcgagcgt cgccagcgtg tcgaggatgt cgaagtcgta gccgtcggcg ctggcgatgt
     4921 agacctgctg gtcgaattga ctgtcgcgca tacacatcgg gccccggggc ccgtcgaacc
     4981 cgacatcgtg cgcggatgcc atcaggtccg gtatctcggg ggagtgggcc cgctggaaga
     5041 tggcctcgag cgcaagcaga ccctcgtaac aggattcggc catcgcgttg agcggtggcg
     5101 cgtcggcgcc gtagcgggcg acgtagctgc ccatcaggtc catcgcaccc gcggtggcca
     5161 gtgaactgaa gtacgccgcg gcgacataga ggttttcggt ggagccggcg ccgctggcca
     5221 gcagcatgtt ctcctccatc agcgggctga accgcgccat gcggtcgtgc ccgccggcgc
     5281 gcgcgaactc gcggttgaac aacacggcgt cctggccgac gagcagcatc aacacggcct
     5341 gcgcccccga cgcgatggcc ttgcggacag gtgcgcggaa atcgtcggtg ccgtacggga
     5401 cgtagatctc ccgtctgagc tcgaggtcca gatctcggca gtacgcgcgg gcggccgcgg
     5461 cggaacggcg cggccagatg tagtcatcgc cgaccaggca ccaggaccgg atgccgaagt
     5521 ggtcgcgcag ccaggcgagc gcgggcgcga tctggatctg cggtgtctcg cctgtgcaga
     5581 acacgcccgg tgtgcgttca ccgccctcgt acaacgaggt gtagacgtac gggatgcggt
     5641 cgcggaccac cggggagatg cggttgcgca cggccgagat gtgccagccg gtcacggcgt
     5701 cgagaccgtg acctcgcaac cggtcggcga cggtccgggc gacgtcgtcg ccgggcgctc
     5761 cgccgtcgag cacctcgatg gtgaccttgc ggccctgcag gccgcctcgg tcgttgacct
     5821 ccttggccgc gagctcggcc acggcctcgc acgaaggcgc gaagattccc gctggccctt
     5881 gaagcggaat caccagcccg acgcggaact caacctcgcc gtcctgcact ccagatcacc
     5941 gtcgatcccg tgtagtctgc gcttcaaagg tttctagcag aaataattca ttctgaacag
     6001 accccgccgt cgacacgagg agacccaaca tggccgccgg acagcagcgc cgccccaacc
     6061 tcctgctgcc gttggtgcgt ctgacccacc tcgcggagtc ggcgatcgaa cgcgtgctcg
     6121 cggactcgtc gctcaagatc gaggactggc gcgtgctcga cgagttggcc ggacggcgca
     6181 ccgtgcccat gagcgatctc gcgcaggcca cgctgatcac gggtccgact ctcaccagaa
     6241 ccgtcgatcg ccttgtgtcg caagggatca tctaccggac tgccgatctg catgaccgcc
     6301 ggcgggtgct cgtggcgttg accccgcggg ggcggacgct gcgcaaccgc ctggtggacg
     6361 cggtagccga ggccgagtgt gcggcttttg aatcgtgcgg gctggacgtc gaccagttgc
     6421 gcgaactcgt cgacaccacc tcgaatttga cttcgtaacc acccgcgccc ggcgcgggcg
     6481 ttcacccttg acttttattt tcatctggat atatttcggg tgaatggaaa ggggtgacca
     6541 tgccgaccta cacattccgt tgttcccact gcggtccctt cgatctcacc tgcgcgatct
     6601 ccgagcgcga tgcggcggcg acctgtccgg agtgccggac gccggcgcgc cgggtcttcg
     6661 gttcggtagg gctgacgaca ttcaccgcgg gacatcaccg cgcattcgac gcggcgtccg
     6721 cgagcgccga aagtcccacg gtggtgaagt cgattcccgc aggcgcggac cgcccgcggg
     6781 ccccgcgccg caatcccggt ctaccgagtc t
//