LOCUS       40924_33522        3114 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pNS2TrcD, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3114)
  AUTHORS   Seleem MN, Anderson B, Sriranganathan N.
  TITLE     Improved Expression Vectors for Bartonella
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3114)
  AUTHORS   Seleem MN, Anderson B, Sriranganathan N.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-MAR-2008) ICTAS, Virginia Tech, 1410 Prices Fork Rd, 
            Blacksburg, VA 24061, USA
REFERENCE   3  (bases 1 to 3114)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3114)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (29-MAR-2008) ICTAS, Virginia Tech, 1410 Prices Fork Rd, Blacksburg,
            VA 24061, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3114
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        28..57
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    65..81
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     misc_feature    98..167
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region
                     (Berg et al., 1989)"
     CDS             218..241
                     /codon_start=1
                     /label=minicistron
                     /note="synthetic cistron containing a ribosome binding site
                     (Shine-Dalgarno sequence), for enhancing the bacterial 
                     expression of a downstream cistron (Schoner, 1997)"
                     /translation="MYRLNKEE"
     CDS             251..268
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             600..1406
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDE"
     CDS             complement(1462..2121)
                     /codon_start=1
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      complement(2122..2891)
                     /direction=LEFT
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
ORIGIN
        1 gtcgaccaga aaaaagatca aaaaaatttg acaattaatc atccggctcg tataatgtgt
       61 ggaattgtga gcggataaca atttcacaca ggaaacagcg ccgctgagaa aaagcgaagc
      121 ggcactgctc tttaacaatt tatcagacaa tctgtgtggg cactcgaccg gaattatcga
      181 ttaactttat tattaaaaat taaagaggta tatattaatg tatcgattaa ataaggagga
      241 ataaaccatg catcatcatc atcatcatgg tggatccgag ctcgagatct gcagctggta
      301 ccatggaatt ccccagtact ccctctagat gcctggcggc agtagcgcgg tggtcccacc
      361 tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc
      421 ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact
      481 gggccttact agaccacgtt gtgtctcaaa atctctgatg ttacattgca caagataaaa
      541 atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta
      601 tgagccatat tcaacgggaa acgtcttgct cgaggccgcg attaaattcc aacatggatg
      661 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
      721 atcgattgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
      781 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc
      841 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga
      901 tccccgggaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
      961 ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
     1021 ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg
     1081 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
     1141 aaatgcataa gcttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
     1201 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
     1261 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
     1321 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
     1381 tgcagtttca tttgatgctc gatgagaact agaatagtct ggaacagcgc acttacgggt
     1441 tgctgcgcaa cccaagtgct accggcgcgg cagcgtgacc cgtgtcggcg gctccaacgg
     1501 ctcgccatcg tccagaaaac acggctcatc gggcatcggc aggcgctgct gcccgcgccg
     1561 ttcccattcc tccgtttcgg tcaaggctgg caggtctggt tccatgcccg gaatgccggg
     1621 ctggctgggc ggctcctcgc cggggccggt cggtagttgc tgctcgcccg gatacagggt
     1681 cgggatgcgg cgcaggtcgc catgccccaa cagcgattcg tcctggtcgt cgtgatcaac
     1741 caccacggcg gcactgaaca ccgacaggcg caactggtcg cggggctggc cccacgccac
     1801 gcggtcattg accacgtagg ccgacacggt gccggggccg ttgagcttca cgacggagat
     1861 ccagcgctcg gccaccaagt ccttgactgc gtattggacc gtccgcaaag aacgtccgat
     1921 gagcttggaa agtgtcttct ggctgaccac cacggcgttc tggtggccca tctgcgccac
     1981 gaggtgatgc agcagcattg ccgccgtggg tttcctcgca ataagcccgg cccacgcctc
     2041 atgcgctttg cgttccgttt gcacccagtg accgggcttg ttcttggctt gaatgccgat
     2101 ttctctggac tgcgtggcca tgcttatctc catgcggtag ggtgccgcac ggttgcggca
     2161 ccatgcgcaa tcagctgcaa cttttcggca gcgcgacaac aattatgcgt tgcgtaaaag
     2221 tggcagtcaa ttacagattt tctttaacct acgcaatgag ctattgcggg gggtgccgca
     2281 atgagctgtt gcgtaccccc cttttttaag ttgttgattt ttaagtcttt cgcatttcgc
     2341 cctatatcta gttctttggt gcccaaagaa gggcacccct gcggggttcc cccacgcctt
     2401 cggcgcggct ccccctccgg caaaaagtgg cccctccggg gcttgttgat cgactgcgcg
     2461 gccttcggcc ttgcccaagg tggcgctgcc cccttggaac ccccgcactc gccgccgtga
     2521 ggctcggggg gcaggcgggc gggcttcgcc ttcgactgcc cccactcgca taggcttggg
     2581 tcgttccagg cgcgtcaagg ccaagccgct gcgcggtcgc tgcgcgagcc ttgacccgcc
     2641 ttccacttgg tgtccaaccg gcaagcgaag cgcgcaggcc gcaggccgga ggcttttccc
     2701 cagagaaaat taaaaaaatt gatggggcaa ggccgcaggc cgcgcagttg gagccggtgg
     2761 gtatgtggtc gaaggctggg tagccggtgg gcaatccctg tggtcaagct cgtgggcagg
     2821 cgcagcctgt ccatcagctt gtccagcagg gttgtccacg ggccgagcga agcgagccag
     2881 ccggtggccg ctcgcggcca tcgtccacat atccacgggc tggcaaggga gcgcagcgac
     2941 cgcgcagggc gaagcccgga gagcaagccc gtagggcgcc gcagccgccg taggcggtca
     3001 cgactttgcg aagcaaagtc tagtgagtat actcaagcat tgagtggccc gccggaggca
     3061 ccgccttgcg ctgcccccgt cgagccggtt ggacaccaaa agggaggggc aggc
//
