LOCUS       40924_41275        4961 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Dual expression shuttle vector pST-2K, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4961)
  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 4961)
  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 4961)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4961)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "2013"; volume: "79"; issue: "5"; pages:
            "1718-1729"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (11-NOV-2012) Signal Transduction Lab-I, National Institute of 
            Immunology, Aruna Asaf Ali Marg, New Delhi, Delhi 110067, India"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: ClustalW v. 2.1
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4961
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    7..308
                     /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 1
                     /note="SD sequence 1"
     CDS             335..352
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      382..425
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     regulatory      383..443
                     /label=transcription terminator 1
                     /note="transcription terminator 1"
                     /regulatory_class="terminator"
     regulatory      455..781
                     /label=UV15 promoter without tetO
                     /note="UV15 promoter without tetO"
                     /regulatory_class="promoter"
     misc_feature    784..790
                     /label=SD sequence 2
                     /note="SD sequence 2"
     misc_feature    791..820
                     /label=MCS2
                     /note="MCS2"
     CDS             821..844
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     terminator      863..905
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      934..980
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             1141..1953
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      2288..2876
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      3043..4936
                     /label=oriM
                     /note="oriM"
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 cataaaacga aaggctcagt cgaaagactg ggcctttcgt
      421 tttatagggc cgcccccttc acccagctga gtactaatat tggatcgtcg gcaccgtcac
      481 ggccgtggga ggcggcacga tccgcgacgt gatgatcggc cgcatcccca cggtgctgcg
      541 cagtgagctc tacgccatcc cggcgttgat ctgtgcgttc gcacgcacag gcccggtgtg
      601 agaagggtct ctgacgagcg ggagaaccca cccggggtgg gcgagtttgt cctgcgtgtg
      661 ctcggtcgag taggctctgg gagtacccgt gtgtacgacc agcacggcat acatcatttc
      721 gacgccgaga gattcgccgc ccgaaatgag cacgatccgc atgcgggccc ggaggaatca
      781 cttcgcaatg gaattcctgc agcacgtgtt aattaagctt gactacaagg acgacgacga
      841 caagtaaaag ctatcagtta actaaaacga aaggctcagt cgaaagactg ggcctttcgt
      901 tttataacta gcgtacgatc gactgccagg catcaaataa aacgaaaggc tcagtcgaaa
      961 gactgggcct ttcgttttat gccatcatgg ccgcggtgat cagctagcca actgacgtcg
     1021 gggggggggg aaagcgacgt tgtgtctcaa aatctctgat gttacattgc acaagataaa
     1081 aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     1141 atgagccata ttcaacggga aacgtcttgc tcgaggccgc gattaaattc caacatggat
     1201 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
     1261 tatcgcttgt atgggaagcc ccatgcgcca gagttgtttc tgaaacatgg caaaggtagc
     1321 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
     1381 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
     1441 atccccggga aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     1501 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     1561 tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg
     1621 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     1681 gaaatgcata atcttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     1741 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     1801 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     1861 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     1921 ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa
     1981 cactggcaga gcattacgct gacttgacgg gacggcggct ttgttgaata aatcgaactt
     2041 ttgctgagtt gaaggatcag atcacgcatc ttcccgacaa cgcagaccgt tccgtggcaa
     2101 agcaaaagtt caaaatcacc aactggtcca cctacaacaa agctctcatc aaccgtggct
     2161 ccctcacttt ctggctggat gatggggcga ttcaggcctg gtatgagtca gcaacacctt
     2221 cttcacgagg cagacctcac tagttccact gagcgtcaga ccccgtagaa aagatcaaag
     2281 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     2341 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     2401 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     2461 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     2521 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     2581 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     2641 gaacgaccta caccgaactg agatacctac agcgtgagca ttgagaaagc gccacgcttc
     2701 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     2761 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     2821 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     2881 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     2941 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     3001 ccgctcgccg cagccgaacg accgagcgca acgcgttacg tagagcccac cagctccgta
     3061 agttcgggtg ctgtgtggct cgtacccgcg cattcaggcg gcagggggtc taacgggtct
     3121 aaggcggcgt gtacggccgc cacagcggct cttagcggcc cggaaacgtc ctcgaaacga
     3181 cgcatgtgtt cctcctggtt ggtacaggtg gttgggggtg ctcggctgtc gctggtgttt
     3241 catcatcagg gctcgacggg agagcggggg agtgtgcagt tgtggggtgg cccctcagcg
     3301 aaatatctga cttggagctc gtgtcggacc atacaccggt gattaatcgt ggtttattat
     3361 caagcgtgag ccacgtcgcc gacgaatttg agcagctctg gctgccgtac tggtccctgg
     3421 caagcgacga tctgctcgag gggatctacc gccaaagccg cgcgtcggcc ctaggccgcc
     3481 ggtacatcga ggcgaaccca acagcgctgg caaacctgct ggtcgtggac gtagaccatc
     3541 cagacgcagc gctccgagcg ctcagcgccc gggggtccca tccgctgccc aacgcgatcg
     3601 tgggcaatcg cgccaacggc cacgcacacg cagtgtgggc actcaacgcc cctgttccac
     3661 gcaccgaata cgcgcggcgt aagccgctcg catacatggc ggcgtgcgcc gaaggccttc
     3721 ggcgcgccgt cgatggcgac cgcagttact caggcctcat gaccaaaaac cccggccaca
     3781 tcgcctggga aacggaatgg ctccactcag atctctacac actcagccac atcgaggccg
     3841 agctcggcgc gaacatgcca ccgccgcgct ggcgtcagca gaccacgtac aaagcggctc
     3901 cgacgccgct agggcggaat tgcgcactgt tcgattccgt caggttgtgg gcctatcttc
     3961 ccgccctcat gcggatctac ctgccgaccc ggaacgtgga cggactcggc cgcgcgatct
     4021 atgccgagtg ccacgcgcga aacgccgaat ttccgtgcaa cgacgtgtgt cccggaccgc
     4081 taccggacag cgaggtccgc gccatcgcca acagcatttg gcgttggatc acaaccaagt
     4141 cgcgcatttg ggcggacggg atcgtggtct acgaggccac actcagtgcg cgccatgcgg
     4201 ccatctcgcg gaagggcgca gcagcgcgca cggcggcgag cacagttgcg cggcgcgcaa
     4261 agtccgcgtc agccatggag gcattgctat gagcgacggc tacagcgacg gctacagcga
     4321 cggctacaac tggcagccga ctgtccgcaa aaagcggcgc gtgaccgccg ccgaaggcgc
     4381 tcgaatcacc ggactatccg aacgccacgt cgtccggctc gtggcgcagg aacgcagcga
     4441 gtggttcgcc gagcaggctg cacgccgcga acgcatccgc gcctatcacg acgacgaggg
     4501 ccactcttgg ccgcaaacgg ccaaacattt cgggctgcat ctggacaccg ttaagcgact
     4561 cggctatcgg gcgaggaaag agcgtgcggc agaacaggaa gcggctcaaa aggcccacaa
     4621 cgaagccgac aatccaccgc tgttctaacg caattgggga gcgggtgtcg cgggggttcc
     4681 gtggggggtt ccgttgcaac gggtcggaca ggtaaaagtc ctggtagacg ctagttttct
     4741 ggtttgggcc atgcctgtct cgttgcgtgt ttcgttgcgt ccgttttgaa taccagccag
     4801 acgagacggg gttctacgaa tcttggtcga taccaagcca tttccgctga atatcgtgga
     4861 gctcaccgcc agaatcggtg gttgtggtga tgtacgtggc gaactccgtt gtagtgcttg
     4921 tggtggcatc cgtggcggta ccgcctgacg ccccccggat c
//
