LOCUS       sequence-69375-e        4565 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for ectopic constitutive expression of mycobacterial proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4565)
  AUTHORS   Parikh A, Kumar D, Chawla Y, Kurthkoti K, Khan S, Varshney U, 
            Nandicoori VK
  TITLE     Development of a new generation of vectors for gene expression, gene
            replacement, and protein-protein interaction studies in 
            mycobacteria.
  JOURNAL   Appl Environ Microbiol. 2013 Mar;79(5):1718-29. doi: 
            10.1128/AEM.03695-12. Epub 2013 Jan 11.
  PUBMED    23315736
REFERENCE   2  (bases 1 to 4565)
  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"; doi: 
            "10.1128/AEM.03695-12"; journalName: "Appl Environ Microbiol"; date:
            "2013-03"; volume: "79"; issue: "5"; pages: "1718-29"
COMMENT     Sequence Label: pST-K
FEATURES             Location/Qualifiers
     source          1..4565
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             335..352
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             425..448
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      complement(467..509)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      538..584
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(734..753)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             745..1560
                     /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(818..837)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1892..2480
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2381..2400
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agtactaata ttggatcgtc ggcaccgtca cggccgtggg aggcggcacg atccgcgacg
       61 tgatgatcgg ccgcatcccc acggtgctgc gcagtgagct ctacgccatc ccggcgttga
      121 tctgtgcgtt cgcacgcaca ggcccggtgt gagaagggtc tctgacgagc gggagaactc
      181 cctatcagtg atagagtttg tcctccctat cagtgataga taggctctgg gagtacccgt
      241 gtgtacgacc agcacggcat acatcatttc gacgccgaga gattcgccgc ccgaaatgag
      301 cacgatccgc atgcggagga atcacttcgc aatgcaccac caccaccacc atatgggatc
      361 cgatatctct agagcggccg cacagctggg gcccgaattc ctgcagcacg tgttaattaa
      421 gcttgactac aaggacgacg acgacaagta aaagctatca gttaactaaa acgaaaggct
      481 cagtcgaaag actgggcctt tcgttttata actagcgtac gatcgactgc caggcatcaa
      541 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatgccatc atggccgcgg
      601 tgatcagcta gccacctgac gtcggggggg ggggaaagcc acgttgtgtc tcaaaatctc
      661 tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac
      721 ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc ttgctcgagg
      781 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
      841 gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agccccatgc gccagagttg
      901 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
      961 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
     1021 gatgcatggt tactcaccac tgcgatcccc gggaaaacag cattccaggt attagaagaa
     1081 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
     1141 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg
     1201 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
     1261 tggcctgttg aacaagtctg gaaagaaatg cataatcttt tgccattctc accggattca
     1321 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     1381 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     1441 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     1501 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     1561 tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc
     1621 ggctttgttg aataaatcga acttttgctg agttgaagga tcagatcacg catcttcccg
     1681 acaacgcaga ccgttccgtg gcaaagcaaa agttcaaaat caccaactgg tccacctaca
     1741 acaaagctct catcaaccgt ggctccctca ctttctggct ggatgatggg gcgattcagg
     1801 cctggtatga gtcagcaaca ccttcttcac gaggcagacc tcactagttc cactgagcgt
     1861 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1921 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1981 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     2041 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     2101 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     2161 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     2221 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     2281 agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     2341 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     2401 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     2461 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     2521 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     2581 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcaacgcgt
     2641 tacgtagagc ccaccagctc cgtaagttcg ggtgctgtgt ggctcgtacc cgcgcattca
     2701 ggcggcaggg ggtctaacgg gtctaaggcg gcgtgtacgg ccgccacagc ggctcttagc
     2761 ggcccggaaa cgtcctcgaa acgacgcatg tgttcctcct ggttggtaca ggtggttggg
     2821 ggtgctcggc tgtcgctggt gtttcatcat cagggctcga cgggagagcg ggggagtgtg
     2881 cagttgtggg gtggcccctc agcgaaatat ctgacttgga gctcgtgtcg gaccatacac
     2941 cggtgattaa tcgtggttta ttatcaagcg tgagccacgt cgccgacgaa tttgagcagc
     3001 tctggctgcc gtactggtcc ctggcaagcg acgatctgct cgaggggatc taccgccaaa
     3061 gccgcgcgtc ggccctaggc cgccggtaca tcgaggcgaa cccaacagcg ctggcaaacc
     3121 tgctggtcgt ggacgtagac catccagacg cagcgctccg agcgctcagc gcccgggggt
     3181 cccatccgct gcccaacgcg atcgtgggca atcgcgccaa cggccacgca cacgcagtgt
     3241 gggcactcaa cgcccctgtt ccacgcaccg aatacgcgcg gcgtaagccg ctcgcataca
     3301 tggcggcgtg cgccgaaggc cttcggcgcg ccgtcgatgg cgaccgcagt tactcaggcc
     3361 tcatgaccaa aaaccccggc cacatcgcct gggaaacgga atggctccac tcagatctct
     3421 acacactcag ccacatcgag gccgagctcg gcgcgaacat gccaccgccg cgctggcgtc
     3481 agcagaccac gtacaaagcg gctccgacgc cgctagggcg gaattgcgca ctgttcgatt
     3541 ccgtcaggtt gtgggcctat cttcccgccc tcatgcggat ctacctgccg acccggaacg
     3601 tggacggact cggccgcgcg atctatgccg agtgccacgc gcgaaacgcc gaatttccgt
     3661 gcaacgacgt gtgtcccgga ccgctaccgg acagcgaggt ccgcgccatc gccaacagca
     3721 tttggcgttg gatcacaacc aagtcgcgca tttgggcgga cgggatcgtg gtctacgagg
     3781 ccacactcag tgcgcgccat gcggccatct cgcggaaggg cgcagcagcg cgcacggcgg
     3841 cgagcacagt tgcgcggcgc gcaaagtccg cgtcagccat ggaggcattg ctatgagcga
     3901 cggctacagc gacggctaca gcgacggcta caactggcag ccgactgtcc gcaaaaagcg
     3961 gcgcgtgacc gccgccgaag gcgctcgaat caccggacta tccgaacgcc acgtcgtccg
     4021 gctcgtggcg caggaacgca gcgagtggtt cgccgagcag gctgcacgcc gcgaacgcat
     4081 ccgcgcctat cacgacgacg agggccactc ttggccgcaa acggccaaac atttcgggct
     4141 gcatctggac accgttaagc gactcggcta tcgggcgagg aaagagcgtg cggcagaaca
     4201 ggaagcggct caaaaggccc acaacgaagc cgacaatcca ccgctgttct aacgcaattg
     4261 gggagcgggt gtcgcggggg ttccgtgggg ggttccgttg caacgggtcg gacaggtaaa
     4321 agtcctggta gacgctagtt ttctggtttg ggccatgcct gtctcgttgc gtgtttcgtt
     4381 gcgtccgttt tgaataccag ccagacgaga cggggttcta cgaatcttgg tcgataccaa
     4441 gccatttccg ctgaatatcg tggagctcac cgccagaatc ggtggttgtg gtgatgtacg
     4501 tggcgaactc cgttgtagtg cttgtggtgg catccgtggc ggtaccgcct gacgcccccc
     4561 ggatc
//
