LOCUS       sequence-69377-e        4954 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for integrative inducible expression of mycobacterial proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4954)
  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 4954)
  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-KiT
FEATURES             Location/Qualifiers
     source          1..4954
                     /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"
     CDS             2789..3412
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(2862..2881)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse 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 tacgtaaata ttggatcgtc gcaccgggtt aagcctggag tgcggtggtg cctggtcggc
     2701 attttcgcag tcgagggctc tcgtgtagcc tgggcgagtt gccgacgcag gcgaccctcc
     2761 tgccacggat cggaggaatc acttcgcaat gtctagatta gataaaagta aagtgattaa
     2821 cagcgcatta gagctgctta atgaggtcgg aatcgaaggt ttaacaaccc gtaaactcgc
     2881 ccagaaactt ggtgtagagc agcctacatt gtattggcat gtaaaaaata agcgagccct
     2941 gctcgacgcc ttagccattg agatgttaga taggcaccat actcactttt gccctttaga
     3001 aggggaaagc tggcaagatt ttttgcgtaa taacgctaaa agttttagat gtgctttact
     3061 aagtcatcgc gatggagcaa aagtacattt aggtacacgg cctacagaaa aacagtatga
     3121 aactctcgaa aatcaattag cctttttatg ccaacaaggt ttttcactag agaacgcgtt
     3181 atatgcactc agcgctgtgg ggcattttac cttaggttgc gtattggaag atcaagagca
     3241 tcaagtcgct aaagaagaaa gggaaacacc tactactgat agtatgccgc cattattacg
     3301 acaagctatc gaattatttg atcaccaagg tgcagagcca gccttcttat tcggccttga
     3361 attgatcatc tgcggattag aaaaacaact taaatgtgaa agtgggtctt aatgaccatg
     3421 ggctacgtat gcgttgcaac cgcgtatgcc caggtcagaa gagtcgcaca agagttgcag
     3481 acccctggaa agaaaaatgg ccagagggcg aaaacaccct ctgaccagcg gagcgggcga
     3541 cgggaatcga acccgcgtag ctagtttgga agaatgggtg tctgccgacc acaaatgggc
     3601 cggtcaagat aggtttttac cccctctcgg ctgcatcctc taagtggaaa gaaattgcag
     3661 gtcgtagaag cgcgttgaag cctgagagtt gcacaggagt tgcaacccgg tagccttgtt
     3721 cacgacgaga ggagacctag atggcacgtc gcggatgggg atcgctgaag actcagcgca
     3781 gcgggaagat ccaagcctca tacgtcaacc cgcaggacgg tgtgaggtac tacgcgttgc
     3841 agacctacga caacaagatg gacgccgaag cctggctcgc gggcgagaag cggctcatcg
     3901 agatggagac ctggacccct ccacaggacc gggcgaagaa ggcagccgcc agcgccatca
     3961 cgctggagga gtacacccgg aagtggctcg tggagcgcga cctcgcagac ggcaccaggg
     4021 atctgtacag cgggcacgcg gagcgccgca tctacccggt gctaggtgaa gtggcggtca
     4081 cagagatgac gccagctctg gtgcgtgcgt ggtgggccgg gatgggtagg aagcacccga
     4141 ctgcccgccg gcatgcctac aacgtcctcc gggcggtgat gaacacagcg gtcgaggaca
     4201 agctgatcgc agagaacccg tgccggatcg agcagaaggc agccgatgag cgcgacgtag
     4261 aggcgctgac gcctgaggag ctggacatcg tcgccgctga gatcttcgag cactaccgga
     4321 tcgcggcata catcctggcg tggacgagcc tccggttcgg agagctgatc gagcttcgcc
     4381 gcaaggacat cgtggacgac ggcatgacga tgaagctccg ggtgcgccgt ggcgcttccc
     4441 gcgtggggaa caagatcgtc gttggcaacg ccaagaccgt ccggtcgaag cgtcctgtga
     4501 cggttccgcc tcacgtcgcg gagatgatcc gagcgcacat gaaggaccgt acgaagatga
     4561 acaagggccc cgaggcattc ctggtgacca cgacgcaggg caaccggctg tcgaagtccg
     4621 cgttcaccaa gtcgctgaag cgtggctacg ccaagatcgg tcggccggaa ctccgcatcc
     4681 acgacctccg cgctgtcggc gctacgttcg ccgctcaggc aggtgcgacg accaaggagc
     4741 tgatggcccg tctcggtcac acgactccta ggatggcgat gaagtaccag atggcgtctg
     4801 aggcccgcga cgaggctatc gctgaggcga tgtccaagct ggccaagacc tcctgaaacg
     4861 caaaaagccc ccctcccaag gacactgagt cctaaagagg ggggtttctt gtcagtacgc
     4921 gaagaaccag gtaccgcctg acgccccccg gatc
//
