pLOI2227 vector (V004961)

LOCUS       40924_28571        3443 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Integration vector pLOI2227, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3443)
  AUTHORS   Martinez-Morales F, Borges AC, Martinez A, Shanmugam KT, Ingram LO.
  TITLE     Chromosomal integration of heterologous DNA in Escherichia coli with
            precise removal of markers and replicons used during construction
  JOURNAL   J. Bacteriol. 181 (22), 7143-7148 (1999)
  PUBMED    10559184
REFERENCE   2  (bases 1 to 3443)
  AUTHORS   Borges AC, Martinez-Morales F, Ingram LO, Shanmugam KT.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-JUL-1999) Microbiology and Cell Science, University of
            Florida, 981 Museum Road, Room 1160, Gainesville, FL 32611-0700, USA
REFERENCE   3  (bases 1 to 3443)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3443)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. 
            Bacteriol."; date: "1999"; volume: "181"; issue: "22"; pages: 
            "7143-7148"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (28-JUL-1999) Microbiology and Cell Science, University of Florida, 
            981 Museum Road, Room 1160, Gainesville, FL 32611-0700, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    17..50
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     promoter        86..104
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    112..182
                     /label=multiple cloning site
                     /note="multiple cloning site"
     protein_bind    214..248
                     /label=flipase binding site
                     /bound_moiety="flipase"
                     /note="flipase-binding site"
     protein_bind    216..249
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     CDS             complement(415..1362)
                     /codon_start=1
                     /label=Rep101(Ts)
                     /note="temperature-sensitive version of the RepA protein
                     needed for replication with the pSC101 origin (Armstrong et
                     al., 1984)"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      complement(1410..1632)
                     /direction=LEFT
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             complement(2501..3292)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"