LOCUS       Exported                4436 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pKfloxANTSAsc-tsOri-pBR, complete sequence.
ACCESSION   JQ301473
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4436)
  AUTHORS   Yoon YG, Koob MD.
  TITLE     Nonreplicating Intracellular Bacterial Vector for Conjugative DNA 
            Transfer into Mitochondria
  JOURNAL   Pharm. Res. (2012) In press
  PUBMED    22350804
REFERENCE   2  (bases 1 to 4436)
  AUTHORS   Koob MD.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-DEC-2011) Lab Medicine and Pathology, ITN and IHG, 
            University of Minnesota, 2101 6th St SE, WMBB, Minneapolis, MN 
            55455, USA
REFERENCE   3  (bases 1 to 4436)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4436
                     /organism="Cloning vector pKfloxANTSAsc-tsOri-pBR"
                     /mol_type="other DNA"
                     /db_xref="taxon:1151787"
     regulatory      44..315
                     /regulatory_class="terminator"
                     /note="phage lambda tl3 terminator"
     terminator      55..301
                     /label=-lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     misc_feature    328..2084
                     /note="pSC101-derived pEL3 temperature sensitive origin 
                     fragment"
     CDS             complement(403..1353)
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /note="pSC101 replication protein; temperature sensitive 
                     RepA, inactive at 42 C"
                     /protein_id="AFC75903.1"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     gene            complement(403..1353)
                     /gene="repA"
                     /label=repA
     rep_origin      1401..1623
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     rep_origin      complement(1421..2084)
                     /direction=LEFT
                     /label=pSC101 ori region
                     /note="pSC101 ori region"
     rep_origin      complement(2109..3045)
                     /direction=LEFT
                     /label=pBR322 replication ori
                     /note="pBR322 replication ori"
     rep_origin      complement(2345..2933)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    complement(3073..3106)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_recomb     3073..3097
                     /label=loxP
                     /note="loxP"
     CDS             complement(3124..3918)
                     /codon_start=1
                     /product="aminoglycoside-3-O-phosphotransferase"
                     /label=aminoglycoside-3-O-phosphotransferase
                     /note="kanamycin resistance"
                     /protein_id="AFC75904.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     protein_bind    complement(4059..4092)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_recomb     4059..4083
                     /label=loxP
                     /note="loxP"
     regulatory      4109..4372
                     /regulatory_class="terminator"
     terminator      4160..4246
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4338..4365
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     misc_feature    4383..3
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"