LOCUS       40924_33292        6207 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pNIT-1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6207)
  AUTHORS   Pandey AK, Raman S, Proff R, Joshi S, Kang CM, Rubin EJ, Husson RN, 
            Sassetti CM.
  TITLE     Nitrile-inducible gene expression in mycobacteria
  JOURNAL   Tuberculosis (Edinb) 89 (1), 12-16 (2009)
  PUBMED    18801704
REFERENCE   2  (bases 1 to 6207)
  AUTHORS   Pandey AK, Proff R, Joshi S, Sassetti CM.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-SEP-2008) Molecular Genetics and Microbiology, 
            University of Massachusetts Medical School, 55 Lake Avenue North. 
            S6-141, Worcester, MA 01655, USA
REFERENCE   3  (bases 1 to 6207)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6207)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Tuberculosis (Edinb)"; date: "2009"; volume: "89"; issue: "1"; 
            pages: "12-16"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (01-SEP-2008) Molecular Genetics and Microbiology, University of 
            Massachusetts Medical School, 55 Lake Avenue North. S6-141, 
            Worcester, MA 01655, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6207
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             141..953
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      1288..1876
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      2035..3930
                     /label=pAL5000 mycobacterial origin of replication
                     /note="pAL5000 mycobacterial origin of replication"
     terminator      complement(3974..4021)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      4059..4173
                     /label=PnitA
                     /note="PnitA"
                     /regulatory_class="promoter"
     misc_feature    4174..4222
                     /label=multiple cloning site
                     /note="multiple cloning site"
     terminator      4229..4268
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     regulatory      4276..4390
                     /label=PnitA
                     /note="PnitA"
                     /regulatory_class="promoter"
     CDS             4391..5350
                     /codon_start=1
                     /gene="nitR"
                     /product="nitrile-inducible regulatory protein"
                     /label=nitR
                     /protein_id="ACI22356.1"
                     /translation="MNTFFSSDQVSAPDRVALWHDVICRSYVPLNITLTSEQPFIGTVS
                     TGNLGTVRIATSSSLPQQITRTRRLIRQDEREYLMVGVQSAGHALVQQHGRTARVGRGG
                     LVFWDTRHPYDILFPTDWRMSVFQFPRYSFGFTEDFIGRMTAVNVGGDRGIGRVVSSFM
                     TSINDATDAGDLAEVASLHNSAVDLLSAAIRTELADQAAASDGLLECVLAYIRQNLADP
                     NLCASQIAAEHNVSVRTLHRLFSATGQGVAEHIRNLRLERIKTELADPTSRRYTISALA
                     RKWGFLDPSTFSRAFKDAYGITAREWAASASASPTEVS"
     gene            4391..5350
                     /gene="nitR"
                     /label=nitR
     terminator      complement(5388..5415)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(5507..5593)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 atgctagcca cctgacgtcg gggggggggg aaagccacgt tgtgtctcaa aatctctgat
       61 gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa
      121 acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcttgc tcgaggccgc
      181 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
      241 ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc ccatgcgcca gagttgtttc
      301 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact
      361 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg
      421 catggttact caccactgcg atccccggga aaacagcatt ccaggtatta gaagaatatc
      481 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga
      541 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat
      601 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc
      661 ctgttgaaca agtctggaaa gaaatgcata atcttttgcc attctcaccg gattcagtcg
      721 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
      781 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga
      841 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg
      901 ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag
      961 aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcggct
     1021 ttgttgaata aatcgaactt ttgctgagtt gaaggatcag atcacgcatc ttcccgacaa
     1081 cgcagaccgt tccgtggcaa agcaaaagtt caaaatcacc aactggtcca cctacaacaa
     1141 agctctcatc aaccgtggct ccctcacttt ctggctggat gatggggcga ttcaggcctg
     1201 gtatgagtca gcaacacctt cttcacgagg cagacctcac tagttccact gagcgtcaga
     1261 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     1321 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1381 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     1441 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1501 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1561 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1621 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagca
     1681 ttgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1741 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1801 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     1861 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     1921 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     1981 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca acgcgtgagc
     2041 ccaccagctc cgtaagttcg ggtgctgtgt ggctcgtacc cgcgcattca ggcggcaggg
     2101 ggtctaacgg gtctaaggcg gcgtgtacgg ccgccacagc ggctcttagc ggcccggaaa
     2161 cgtcctcgaa acgacgcatg tgttcctcct ggttggtaca ggtggttggg ggtgctcggc
     2221 tgtcgctggt gtttcatcat cagggctcga cgggagagcg ggggagtgtg cagttgtggg
     2281 gtggcccctc agcgaaatat ctgacttgga gctcgtgtcg gaccatacac cggtgattaa
     2341 tcgtggttta ttatcaagcg tgagccacgt cgccgacgaa tttgagcagc tctggctgcc
     2401 gtactggtcc ctggcaagcg acgatctgct cgaggggatc taccgccaaa gccgcgcgtc
     2461 ggccctaggc cgccggtaca tcgaggcgaa cccaacagcg ctggcaaacc tgctggtcgt
     2521 ggacgtagac catccagacg cagcgctccg agcgctcagc gcccgggggt cccatccgct
     2581 gcccaacgcg atcgtgggca atcgcgccaa cggccacgca cacgcagtgt gggcactcaa
     2641 cgcccctgtt ccacgcaccg aatacgcgcg gcgtaagccg ctcgcataca tggcggcgtg
     2701 cgccgaaggc cttcggcgcg ccgtcgatgg cgaccgcagt tactcaggcc tcatgaccaa
     2761 aaaccccggc cacatcgcct gggaaacgga atggctccac tcagatctct acacactcag
     2821 ccacatcgag gccgagctcg gcgcgaacat gccaccgccg cgctggcgtc agcagaccac
     2881 gtacaaagcg gctccgacgc cgctagggcg gaattgcgca ctgttcgatt ccgtcaggtt
     2941 gtgggcctat cttcccgccc tcatgcggat ctacctgccg acccggaacg tggacggact
     3001 cggccgcgcg atctatgccg agtgccacgc gcgaaacgcc gaatttccgt gcaacgacgt
     3061 gtgtcccgga ccgctaccgg acagcgaggt ccgcgccatc gccaacagca tttggcgttg
     3121 gatcacaacc aagtcgcgca tttgggcgga cgggatcgtg gtctacgagg ccacactcag
     3181 tgcgcgccat gcggccatct cgcggaaggg cgcagcagcg cgcacggcgg cgagcacagt
     3241 tgcgcggcgc gcaaagtccg cgtcagccat ggaggcattg ctatgagcga cggctacagc
     3301 gacggctaca gcgacggcta caactggcag ccgactgtcc gcaaaaagcg gcgcgtgacc
     3361 gccgccgaag gcgctcgaat caccggacta tccgaacgcc acgtcgtccg gctcgtggcg
     3421 caggaacgca gcgagtggtt cgccgagcag gctgcacgcc gcgaacgcat ccgcgcctat
     3481 cacgacgacg agggccactc ttggccgcaa acggccaaac atttcgggct gcatctggac
     3541 accgttaagc gactcggcta tcgggcgagg aaagagcgtg cggcagaaca ggaagcggct
     3601 caaaaggccc acaacgaagc cgacaatcca ccgctgttct aacgcaattg gggagcgggt
     3661 gtcgcggggg ttccgtgggg ggttccgttg caacgggtcg gacaggtaaa agtcctggta
     3721 gacgctagtt ttctggtttg ggccatgcct gtctcgttgc gtgtttcgtt gcgtccgttt
     3781 tgaataccag ccagacgaga cggggttcta cgaatcttgg tcgataccaa gccatttccg
     3841 ctgaatatcg tggagctcac cgccagaatc ggtggttgtg gtgatgtacg tggcgaactc
     3901 cgttgtagtg cttgtggtgg catccgtggc gcggccgcgg taccagatct ccggatatag
     3961 ttcctccttt cagcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta
     4021 gttattgctc agttaactac gtcgacatcg atagtactgc gaactccctt atgcgggtgg
     4081 cgcagaatgc caggaccctt gtcattccac gtcaattcat gcgccttttc acctcgtact
     4141 gtcctgccaa acacaagcaa cggaggtacg catatggata tcgaattcaa gcttttcgaa
     4201 ccatggttaa ttaaatttaa attaaaccga tacaattaaa ggctcctttt ggagcctttt
     4261 tttttggagc ttaaggcgaa ctcccttatg cgggtggcgc agaatgccag gacccttgtc
     4321 attccacgtc aattcatgcg ccttttcacc tcgtactgtc ctgccaaaca caagcaacgg
     4381 aggtacggac atgaacactt tcttctcctc agaccaggtc tcggcgcccg atcgcgtcgc
     4441 gctctggcac gatgtcatct gccgtagcta tgtcccgctc aacatcaccc tcacgagcga
     4501 gcaacccttc atcggtacgg tctcgacggg caacttgggc acggtacgta tcgcgacgtc
     4561 ctcgtcactg ccccaacaga tcacccgcac tcgtcgcttg atcaggcagg acgagcgtga
     4621 gtacctcatg gttggggtgc agtccgccgg ccatgcactc gtgcagcagc acggcagaac
     4681 tgcacgagtc ggtcgcggtg gactggtctt ctgggacacc cgccatccct acgacatcct
     4741 cttcccgaca gactggagga tgagcgtatt ccagttcccg cgatactctt tcggcttcac
     4801 cgaagacttc atcggcagga tgaccgcggt gaacgtcggg ggcgatcgcg gtatcggccg
     4861 agtggtttca tccttcatga caagcatcaa cgatgcgacc gacgcaggag acttggcgga
     4921 ggtagcttca ctccacaaca gtgctgtcga tcttctgtca gcggcgatac ggaccgagct
     4981 tgccgatcaa gccgccgcct ccgacggcct actcgagtgt gtgctcgcgt atatccgaca
     5041 gaacctggcc gacccgaacc tgtgtgcctc acagatcgcg gcggaacaca acgtctctgt
     5101 gcggaccctc caccgactgt tctcggccac gggacagggc gtggccgaac acatccgtaa
     5161 cctccgactc gagcgcatca agactgagct ggcagaccca acgagccggc gatatacgat
     5221 cagcgctttg gcgagaaaat gggggttcct cgatccctca acgttctcac gcgcgttcaa
     5281 agacgcctac ggcatcactg cccgagagtg ggcggcttct gcatcagcat caccgacgga
     5341 ggtttcgtag tctagatatg acgacaggaa gagtttgtag aaacgcaaaa aggccatccg
     5401 tcaggatggc cttctgctta atttgatgcc tggcagttta tggcgggcgt cctgcccgcc
     5461 accctccggg ccgttgcttc gcaacgttca aatccgctcc cggcggattt gtcctactca
     5521 ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtctttc gactgagcct
     5581 ttcgttttat ttgatgcctg gcagttccct actctcgcat ggggagaccc cacactacca
     5641 tcggcgctag agccgtgaac gacagggcga acgccagccc gccgacggcg agggttccga
     5701 ccgctgcaac tcccggtgca accttgtccc ggtctattct cttcactgca ccagctccaa
     5761 tctggtgtga atgcccctcg tctgttcgcg caggcggggg gctctattcg tttgtcagca
     5821 tcgaaagtag ccagatcagg gatgcgttgc aaccgcgtat gcccaggtca gaagagtcgc
     5881 acaagagttg cagacccctg gaaagaaaaa tggccagagg gcgaaaacac cctctgacca
     5941 gcggagcggg cgacgggaat cgaacccgcg tagctagttt ggaagaatgg gtgtctgccg
     6001 accacatata tgggccggtc aagataggtt tttaccccct ctcggctgca tcctctaagt
     6061 ggaaagaaat tgcaggtcgt agaagcgcgt tgaagcctga gagttgcaca ggagttgcaa
     6121 cccggtagcc ttgttcacga cgagaggaga cctagttggc acgtcgcgga tggggatcgc
     6181 tgaagactca gcgcagcggg aggatca
//
