LOCUS       Exported                4954 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Integrative inducible shuttle vector pST-KIT, complete sequence.
ACCESSION   KC153035
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. 79 (5), 1718-1729 (2013)
  PUBMED    23315736
REFERENCE   2  (bases 1 to 4954)
  AUTHORS   Parikh A, Kumar D, Chawla Y, Kurthkoti K, Khan S, Varshney U, 
            Nandicoori VK.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-NOV-2012) Signal Transduction Lab-I, National 
            Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, Delhi 
            110067, India
REFERENCE   3  (bases 1 to 4954)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: ClustalW v. 2.1
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4954
                     /organism="Integrative inducible shuttle vector pST-KIT"
                     /mol_type="other DNA"
                     /db_xref="taxon:1283731"
     regulatory      7..308
                     /regulatory_class="promoter"
                     /note="Pmyc1tetO; UV15 promoter with tetO"
     protein_bind    179..196
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    315..321
                     /label=SD sequence
                     /note="SD sequence"
     misc_feature    332..352
                     /label=His tag
                     /note="His tag"
     CDS             335..352
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    349..424
                     /label=MCS
                     /note="MCS"
     misc_feature    425..452
                     /label=FLAG tag
                     /note="FLAG tag"
     CDS             425..448
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    462..579
                     /label=transcription terminator
                     /note="transcription terminator"
     terminator      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"
     CDS             745..1560
                     /codon_start=1
                     /product="kanamycin resistance protein"
                     /label=kanamycin resistance protein
                     /protein_id="AGE45663.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPHAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHNLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      1892..2480
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     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"
     CDS             3741..4856
                     /codon_start=1
                     /product="integrase"
                     /label=integrase
                     /protein_id="AGE45664.1"
                     /translation="MARRGWGSLKTQRSGKIQASYVNPQDGVRYYALQTYDNKMDAEAW
                     LAGEKRLIEMETWTPPQDRAKKAAASAITLEEYTRKWLVERDLADGTRDLYSGHAERRI
                     YPVLGEVAVTEMTPALVRAWWAGMGRKHPTARRHAYNVLRAVMNTAVEDKLIAENPCRI
                     EQKAADERDVEALTPEELDIVAAEIFEHYRIAAYILAWTSLRFGELIELRRKDIVDDGM
                     TMKLRVRRGASRVGNKIVVGNAKTVRSKRPVTVPPHVAEMIRAHMKDRTKMNKGPEAFL
                     VTTTQGNRLSKSAFTKSLKRGYAKIGRPELRIHDLRAVGATFAAQAGATTKELMARLGH
                     TTPRMAMKYQMASEARDEAIAEAMSKLAKTS"