LOCUS       Exported                3443 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Integration vector pLOI2227, complete sequence.
ACCESSION   AF172937
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3443
                     /organism="Integration vector pLOI2227"
                     /mol_type="other DNA"
                     /db_xref="taxon:108603"
     protein_bind    16..50
                     /label=flipase binding site
                     /bound_moiety="flipase"
                     /note="flipase-binding site"
     protein_bind    17..50
                     /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)"
     promoter        86..104
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    103
                     /label=T7 RNA pol transcription initiation site
                     /note="T7 RNA pol transcription initiation site"
     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(412..1362)
                     /codon_start=1
                     /product="Rep ts replication protein"
                     /label=Rep ts replication protein
                     /protein_id="AAF79921.1"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      1362..1742
                     /label=replication origin from plasmid pHS1
                     /note="replication origin from plasmid pHS1"
     rep_origin      1410..1632
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             complement(2498..3292)
                     /codon_start=1
                     /gene="kan"
                     /product="kanamicin-resistance protein"
                     /label=kan
                     /protein_id="AAF13161.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(2498..3292)
                     /gene="kan"
                     /label=kan