pKfloxANTSAsc-tsOri-pBR vector (V005245)

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LOCUS       40924_26664        4436 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pKfloxANTSAsc-tsOri-pBR, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4436)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Pharm. Res.
            (2012) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (16-DEC-2011) Lab Medicine and Pathology, ITN and IHG, University of
            Minnesota, 2101 6th St SE, WMBB, Minneapolis, MN 55455, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4436
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      55..301
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     CDS             complement(406..1353)
                     /codon_start=1
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      complement(1401..1623)
                     /direction=LEFT
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     rep_origin      complement(2345..2933)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    3073..3106
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(3127..3918)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     terminator      4160..4246
                     /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    complement(join(4383..4436,1..3))
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"