LOCUS       sequence-399289-        5470 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for ectopic inducible expression of mycobacterial proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5470)
  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 5470)
  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-KT
FEATURES             Location/Qualifiers
     source          1..5470
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(98..197)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             542..559
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             632..655
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      complement(674..716)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      745..791
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(941..960)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             952..1767
                     /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(1025..1044)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2098..2686
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2587..2606
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2995..3618
                     /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(3068..3087)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
ORIGIN
        1 atgtacgtgg cgaactccgt tgtagtgcct gtggtggcat ccgtggcggt accgcctgac
       61 gccccccgga tcagtactag taagggtggg cgcgccgacc cagctttctt gtacaaagtt
      121 ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa
      181 aataaaatca ttatttgcca tccagctgat actaatattg gatcgtcggc accgtcacgg
      241 ccgtgggagg cggcacgatc cgcgacgtga tgatcggccg catccccacg gtgctgcgca
      301 gtgagctcta cgccatcccg gcgttgatct gtgcgttcgc acgcacaggc ccggtgtgag
      361 aagggtctct gacgagcggg agaactccct atcagtgata gagtttgtcc tccctatcag
      421 tgatagatag gctctgggag tacccgtgtg tacgaccagc acggcataca tcatttcgac
      481 gccgagagat tcgccgcccg aaatgagcac gatccgcatg cggaggaatc acttcgcaat
      541 gcaccaccac caccaccata tgggatccga tatctctaga gcggccgcac agctggggcc
      601 cgaattcctg cagcacgtgt taattaagct tgactacaag gacgacgacg acaagtaaaa
      661 gctatcagtt aactaaaacg aaaggctcag tcgaaagact gggcctttcg ttttataact
      721 agcgtacgat cgactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
      781 tttcgtttta tgccatcatg gccgcggtga tcagctagcc acctgacgtc gggggggggg
      841 gaaagccacg ttgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca
      901 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat
      961 attcaacggg aaacgtcttg ctcgaggccg cgattaaatt ccaacatgga tgctgattta
     1021 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg
     1081 tatgggaagc cccatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat
     1141 gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc
     1201 atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccggg
     1261 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg
     1321 ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc
     1381 gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg
     1441 agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat
     1501 aatcttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac
     1561 cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca
     1621 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta
     1681 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt
     1741 catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag
     1801 agcattacgc tgacttgacg ggacggcggc tttgttgaat aaatcgaact tttgctgagt
     1861 tgaaggatca gatcacgcat cttcccgaca acgcagaccg ttccgtggca aagcaaaagt
     1921 tcaaaatcac caactggtcc acctacaaca aagctctcac caaccgtggc tccctcactt
     1981 tctggctgga tgatggggcg attcaggcct ggtatgagtc agcaacacct tcttcacgag
     2041 gcagacctca ctagttccat gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2101 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2161 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2221 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     2281 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2341 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2401 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     2461 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaggggag
     2521 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2581 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2641 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2701 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2761 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2821 cagccgaacg accgagcgca acgcgttacg taaatattgg atcgtcgcac cgggttaagc
     2881 ctggagtgcg gtggtgcctg gtcggcattt tcgcagtcga gggctctcgt gtagcctggg
     2941 cgagttgccg acgcaggcga ccctcctgcc acggatcgga ggaatcactt cgcaatgtca
     3001 agattagata aaagtaaagt gattaacagc gcattagagc tgcttaatga ggtcggaatc
     3061 gaaggtttaa caacccgtaa actcgcccag aaacttggtg tagagcagcc tacattgtat
     3121 tggcatgtaa aaaataagcg agccctgctc gacgccttag ccattgagat gttagatagg
     3181 caccatactc acttttgccc tttagaaggg gaaagctggc aagatttttt gcgtaataac
     3241 gctaaaagtt ttagatgtgc tttactaagt catcgcgatg gagcaaaagt acatttaggt
     3301 acacggccta cagaaaaaca gtatgaaact ctcgaaaatc aattagcctt tttatgccaa
     3361 caaggttttt cactagagaa cgcgttatat gcactcagcg ctgtggggca ttttacctta
     3421 ggttgcgtat tggaagatca agagcatcaa gtcgctaaag aagaaaggga aacacctact
     3481 actgatagta tgccgccatt attacgacaa gctatcgaat tatttgatca ccaaggtgca
     3541 gagccagcct tcttattcgg ccttgaattg atcatctgcg gattagaaaa acaacttaaa
     3601 tgtgaaagtg ggtcttaatg accatgggct acgtagagcc caccagctcc gtaagttcgg
     3661 gcgctgtgtg gctcgtaccc gcgcattcag gcggcagggg gtctaacggg tctaaggcgg
     3721 cgtgtacggc cgccacagcg gctctcagcg gcccggaaac gtcctcgaaa cgacgcatgt
     3781 gttcctcctg gttggtacag gtggttgggg gtgctcggct gtcgctggtg ttccaccacc
     3841 agggctcgac gggagagcgg gggagtgtgc agttgtgggg tggcccctca gcgaaatatc
     3901 tgacttggag ctcgtgtcgg accatacacc ggtgattaat cgtggtctac taccaagcgt
     3961 gagccacgtc gccgacgaat ttgagcagct ctggctgccg tactggccgc tggcaagcga
     4021 cgatctgctc gaggggatct accgccaaag ccgcgcgtcg gccctaggcc gccggtacat
     4081 cgaggcgaac ccaacagcgc tggcaaacct gctggtcgtg gacgtagacc atccagacgc
     4141 agcgctccga gcgctcagcg cccgggggtc ccatccgctg cccaacgcga tcgtgggcaa
     4201 tcgcgccaac ggccacgcac acgcagtgtg ggcactcaac gcccctgttc cacgcaccga
     4261 atacgcgcgg cgtaagccgc tcgcatacat ggcggcgtgc accgaaggcc ttcggcgcgc
     4321 cgtcgatggc gaccgcagtt actcaggcct catgaccaaa aaccccggcc acatcgcctg
     4381 ggaaacggaa tggctccact cagatctcta cacactcagc cacatcgagg ccgagctcgg
     4441 cgcgaacatg ccaccgccgc gctggcgtca gcagaccacg tacaaagcgg ctccgacgcc
     4501 gctagggcgg aattgcgcac tgttcgattc cgtcaggttg tgggcctatc gtcccgccct
     4561 catgcggatc tacctgccga cccggaacgt ggacggactc ggccgcgcga tctatgccga
     4621 gtgccacgcg cgaaacgccg aatttccgtg caacgacgtg tgtcccggac cgctaccgga
     4681 cagcgaggtc cgcgccatcg ccaacagcat ttggcgttgg atcacaacca agtcgcgcat
     4741 ttgggcggac gggatcgtgg tctacgaggc cacactcagt gcgcgccagt cggccatctc
     4801 gcggaagggc gcagcagcgc gcacggcggc gagcacagtt gcgcggcgcg caaagtccgc
     4861 gtcagccatg gaggcattgc tatgagcgac ggctacagcg acggctacaa ccggcagccg
     4921 actgtccgca aaaagcggcg cgtgaccgcc gccgaaggcg ctcgaatcac cggactatcc
     4981 gaacgccacg tcgtccggct cgtggcgcag gaacgcagcg agtggctcgc cgagcaggct
     5041 gcacgccgcg aacgcatccg cgcctatcac gacgacgagg gccactcttg gccgcaaacg
     5101 gccaaacatt tcgggctgca tctggacacc gttaagcgac tcggctatcg ggcgaggaaa
     5161 gagcgtgcgg cagaacagga agcggctcaa aaggcccaca acgaagccga caatccaccg
     5221 ctgttctaac gcaattgggg agcgggtgtc gcgggggttt cgtggggggt tccgttgcaa
     5281 cgggtcggac aggtaaaagt cctggtagac gctagttttc tggtttgggc catgcctgtc
     5341 tcgttgcgtg tttcgttgcg cccgttttga ataccagcca gacgagacgg ggttctacga
     5401 atcttggtcg ataccaagcc atttccgctg aatatcgggg agctcaccgc cagaatcggt
     5461 ggttgtggtg
//
