LOCUS       40924_33517        3115 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pNS2TrcB, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3115)
  AUTHORS   Seleem MN, Anderson B, Sriranganathan N.
  TITLE     Improved Expression Vectors for Bartonella
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3115)
  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 3115)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3115)
  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            "Unpublished"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..3115
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        29..58
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    66..82
                     /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    99..168
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region
                     (Berg et al., 1989)"
     CDS             219..242
                     /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             252..269
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             601..1407
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDE"
     CDS             complement(1463..2122)
                     /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(2123..2892)
                     /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 gtcgacggaa aatttttttt caaaagtatt gacaattaat catccggctc gtataatgtg
       61 tggaattgtg agcggataac aatttcacac aggaaacagc gccgctgaga aaaagcgaag
      121 cggcactgct ctttaacaat ttatcagaca atctgtgtgg gcactcgacc ggaattatcg
      181 attaacttta ttattaaaaa ttaaagaggt atatattaat gtatcgatta aataaggagg
      241 aataaaccat gcatcatcat catcatcatg gtggatccga gctcgagatc tgcagctggt
      301 accatggaat tccccagtac tccctctaga tgcctggcgg cagtagcgcg gtggtcccac
      361 ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc
      421 cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac
      481 tgggccttac tagaccacgt tgtgtctcaa aatctctgat gttacattgc acaagataaa
      541 aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
      601 atgagccata ttcaacggga aacgtcttgc tcgaggccgc gattaaattc caacatggat
      661 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
      721 tatcgattgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
      781 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
      841 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
      901 atccccggga aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
      961 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     1021 tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg
     1081 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     1141 gaaatgcata agcttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     1201 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     1261 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     1321 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     1381 ttgcagtttc atttgatgct cgatgagaac tagaatagtc tggaacagcg cacttacggg
     1441 ttgctgcgca acccaagtgc taccggcgcg gcagcgtgac ccgtgtcggc ggctccaacg
     1501 gctcgccatc gtccagaaaa cacggctcat cgggcatcgg caggcgctgc tgcccgcgcc
     1561 gttcccattc ctccgtttcg gtcaaggctg gcaggtctgg ttccatgccc ggaatgccgg
     1621 gctggctggg cggctcctcg ccggggccgg tcggtagttg ctgctcgccc ggatacaggg
     1681 tcgggatgcg gcgcaggtcg ccatgcccca acagcgattc gtcctggtcg tcgtgatcaa
     1741 ccaccacggc ggcactgaac accgacaggc gcaactggtc gcggggctgg ccccacgcca
     1801 cgcggtcatt gaccacgtag gccgacacgg tgccggggcc gttgagcttc acgacggaga
     1861 tccagcgctc ggccaccaag tccttgactg cgtattggac cgtccgcaaa gaacgtccga
     1921 tgagcttgga aagtgtcttc tggctgacca ccacggcgtt ctggtggccc atctgcgcca
     1981 cgaggtgatg cagcagcatt gccgccgtgg gtttcctcgc aataagcccg gcccacgcct
     2041 catgcgcttt gcgttccgtt tgcacccagt gaccgggctt gttcttggct tgaatgccga
     2101 tttctctgga ctgcgtggcc atgcttatct ccatgcggta gggtgccgca cggttgcggc
     2161 accatgcgca atcagctgca acttttcggc agcgcgacaa caattatgcg ttgcgtaaaa
     2221 gtggcagtca attacagatt ttctttaacc tacgcaatga gctattgcgg ggggtgccgc
     2281 aatgagctgt tgcgtacccc ccttttttaa gttgttgatt tttaagtctt tcgcatttcg
     2341 ccctatatct agttctttgg tgcccaaaga agggcacccc tgcggggttc ccccacgcct
     2401 tcggcgcggc tccccctccg gcaaaaagtg gcccctccgg ggcttgttga tcgactgcgc
     2461 ggccttcggc cttgcccaag gtggcgctgc ccccttggaa cccccgcact cgccgccgtg
     2521 aggctcgggg ggcaggcggg cgggcttcgc cttcgactgc ccccactcgc ataggcttgg
     2581 gtcgttccag gcgcgtcaag gccaagccgc tgcgcggtcg ctgcgcgagc cttgacccgc
     2641 cttccacttg gtgtccaacc ggcaagcgaa gcgcgcaggc cgcaggccgg aggcttttcc
     2701 ccagagaaaa ttaaaaaaat tgatggggca aggccgcagg ccgcgcagtt ggagccggtg
     2761 ggtatgtggt cgaaggctgg gtagccggtg ggcaatccct gtggtcaagc tcgtgggcag
     2821 gcgcagcctg tccatcagct tgtccagcag ggttgtccac gggccgagcg aagcgagcca
     2881 gccggtggcc gctcgcggcc atcgtccaca tatccacggg ctggcaaggg agcgcagcga
     2941 ccgcgcaggg cgaagcccgg agagcaagcc cgtagggcgc cgcagccgcc gtaggcggtc
     3001 acgactttgc gaagcaaagt ctagtgagta tactcaagca ttgagtggcc cgccggaggc
     3061 accgccttgc gctgcccccg tcgagccggt tggacaccaa aagggagggg caggc
//
