pS238D.NIa vector (V016153)

LOCUS       62056_19140        7255 bp DNA     circular SYN 26-NOV-2019
DEFINITION  Expression vector pS238D.NIa, complete sequence.
ACCESSION   MN393462
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7255)
  AUTHORS   Calles B, Goni-Moreno A, de Lorenzo V.
  TITLE     Digitalizing heterologous gene expression in Gram-negative bacteria 
            with a portable on/off module
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7255)
  AUTHORS   Calles B, Goni-Moreno A, de Lorenzo V.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-AUG-2019) Systems Biology Program, Centro Nacional de 
            Biotecnologia, Darwin, 3, Cantoblanco, Madrid 28049, Spain
REFERENCE   3  (bases 1 to 7255)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (28-AUG-2019) Systems Biology Program, Centro Nacional de 
            Biotecnologia, Darwin, 3, Cantoblanco, Madrid 28049, Spain"
FEATURES             Location/Qualifiers
     source          1..7255
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      5..99
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             231..1043
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     oriT            1213..1321
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(1335..1994)
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     rep_origin      complement(1995..2766)
                     /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"
     terminator      complement(2881..2967)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(2995..3957)
                     /label=XylS
                     /note="XylS regulator encoded by the Pseudomonas putida TOL
                     plasmid pWWO"
     promoter        4857..4937
                     /label=Pm promoter
                     /note="The bacterial Pm promoter is activated by XylS in
                     the presence of benzoate or m-toluate (Marques et al., 
                     1999)."
     regulatory      4972..4978
                     /regulatory_class="ribosome_binding_site"
     CDS             4990..6069
                     /label=lacI
                     /note="lac repressor"
     CDS             6070..6087
                     /label=6xHis
                     /note="6xHis affinity tag"
     gene            6107..6847
                     /gene="nIa"
                     /label=nIa
     CDS             6107..6847
                     /codon_start=1
                     /transl_table=11
                     /gene="nIa"
                     /product="NIa"
                     /label=nIa
                     /note="specific protease"
                     /protein_id="QGN03492.1"
                     /translation="MASSKSLFRGLRDYNPIASSICQLNNSSGARQSVMFGLGFGGLIV
                     TNQHLFKRNDGELTIRSHHGEFVVKDTKTLKLLPCKGRDIVIIRLPKDFPPFPKRLQFR
                     TPTTEDRVCLIGSNFQTKSISSTMSETSATYPVDNSHFWKHWISTKDGHCGLPIVSTRD
                     GSILGLHSLANSTNTQNFYAAFPDNFETTYLSNQDNDNWIKQWRYNPDEVCWGSLQLKR
                     DIPQSPFTICKLLTDLDGEFVYTQ"
     regulatory      6925..6954
                     /label=synthetic T500 terminator
                     /note="synthetic T500 terminator"
                     /regulatory_class="terminator"
     terminator      complement(6961..6988)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(7028..7074)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     regulatory      complement(7090..7197)
                     /note="anti LacI sRNA; MicC-based"
                     /regulatory_class="other"
     regulatory      complement(7198..7255)
                     /regulatory_class="promoter"