LOCUS       40924_33512        3109 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pNS2Trc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3109)
  AUTHORS   Gillaspie D, Perkins I, Larsen K, McCord A, Pangonis S, Sweger D, 
            Seleem MN, Sriranganathan N, Anderson BE.
  TITLE     Plasmid-based system for high-level gene expression and antisense 
            gene knockdown in Bartonella henselae
  JOURNAL   Appl. Environ. Microbiol. 75 (16), 5434-5436 (2009)
  PUBMED    19542333
REFERENCE   2  (bases 1 to 3109)
  AUTHORS   Seleem MN, Anderson B, Sriranganathan N.
  TITLE     Improved Expression Vectors for Bartonella
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 3109)
  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   4  (bases 1 to 3109)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 3109)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "2009"; volume: "75"; issue: "16"; 
            pages: "5434-5436"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (29-MAR-2008) ICTAS, Virginia Tech, 1410 Prices Fork Rd, Blacksburg,
            VA 24061, USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3109
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        23..52
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    60..76
                     /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    93..162
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region
                     (Berg et al., 1989)"
     CDS             213..236
                     /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             246..263
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             595..1401
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDE"
     CDS             complement(1457..2116)
                     /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(2117..2886)
                     /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 gtcgacattc tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat
       61 tgtgagcgga taacaatttc acacaggaaa cagcgccgct gagaaaaagc gaagcggcac
      121 tgctctttaa caatttatca gacaatctgt gtgggcactc gaccggaatt atcgattaac
      181 tttattatta aaaattaaag aggtatatat taatgtatcg attaaataag gaggaataaa
      241 ccatgcatca tcatcatcat catggtggat ccgagctcga gatctgcagc tggtaccatg
      301 gaattcccca gtactccctc tagatgcctg gcggcagtag cgcggtggtc ccacctgacc
      361 ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg tctccccatg
      421 cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
      481 ttactagacc acgttgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata
      541 tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
      601 catattcaac gggaaacgtc ttgctcgagg ccgcgattaa attccaacat ggatgctgat
      661 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcga
      721 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
      781 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
      841 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
      901 gggaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
      961 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     1021 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     1081 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     1141 cataagcttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     1201 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     1261 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     1321 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     1381 tttcatttga tgctcgatga gaactagaat agtctggaac agcgcactta cgggttgctg
     1441 cgcaacccaa gtgctaccgg cgcggcagcg tgacccgtgt cggcggctcc aacggctcgc
     1501 catcgtccag aaaacacggc tcatcgggca tcggcaggcg ctgctgcccg cgccgttccc
     1561 attcctccgt ttcggtcaag gctggcaggt ctggttccat gcccggaatg ccgggctggc
     1621 tgggcggctc ctcgccgggg ccggtcggta gttgctgctc gcccggatac agggtcggga
     1681 tgcggcgcag gtcgccatgc cccaacagcg attcgtcctg gtcgtcgtga tcaaccacca
     1741 cggcggcact gaacaccgac aggcgcaact ggtcgcgggg ctggccccac gccacgcggt
     1801 cattgaccac gtaggccgac acggtgccgg ggccgttgag cttcacgacg gagatccagc
     1861 gctcggccac caagtccttg actgcgtatt ggaccgtccg caaagaacgt ccgatgagct
     1921 tggaaagtgt cttctggctg accaccacgg cgttctggtg gcccatctgc gccacgaggt
     1981 gatgcagcag cattgccgcc gtgggtttcc tcgcaataag cccggcccac gcctcatgcg
     2041 ctttgcgttc cgtttgcacc cagtgaccgg gcttgttctt ggcttgaatg ccgatttctc
     2101 tggactgcgt ggccatgctt atctccatgc ggtagggtgc cgcacggttg cggcaccatg
     2161 cgcaatcagc tgcaactttt cggcagcgcg acaacaatta tgcgttgcgt aaaagtggca
     2221 gtcaattaca gattttcttt aacctacgca atgagctatt gcggggggtg ccgcaatgag
     2281 ctgttgcgta cccccctttt ttaagttgtt gatttttaag tctttcgcat ttcgccctat
     2341 atctagttct ttggtgccca aagaagggca cccctgcggg gttcccccac gccttcggcg
     2401 cggctccccc tccggcaaaa agtggcccct ccggggcttg ttgatcgact gcgcggcctt
     2461 cggccttgcc caaggtggcg ctgccccctt ggaacccccg cactcgccgc cgtgaggctc
     2521 ggggggcagg cgggcgggct tcgccttcga ctgcccccac tcgcataggc ttgggtcgtt
     2581 ccaggcgcgt caaggccaag ccgctgcgcg gtcgctgcgc gagccttgac ccgccttcca
     2641 cttggtgtcc aaccggcaag cgaagcgcgc aggccgcagg ccggaggctt ttccccagag
     2701 aaaattaaaa aaattgatgg ggcaaggccg caggccgcgc agttggagcc ggtgggtatg
     2761 tggtcgaagg ctgggtagcc ggtgggcaat ccctgtggtc aagctcgtgg gcaggcgcag
     2821 cctgtccatc agcttgtcca gcagggttgt ccacgggccg agcgaagcga gccagccggt
     2881 ggccgctcgc ggccatcgtc cacatatcca cgggctggca agggagcgca gcgaccgcgc
     2941 agggcgaagc ccggagagca agcccgtagg gcgccgcagc cgccgtaggc ggtcacgact
     3001 ttgcgaagca aagtctagtg agtatactca agcattgagt ggcccgccgg aggcaccgcc
     3061 ttgcgctgcc cccgtcgagc cggttggaca ccaaaaggga ggggcaggc
//
