LOCUS       Exported                5819 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pL0B_0-R, complete sequence.
ACCESSION   KY132037
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5819)
  AUTHORS   Hochrein L, Machens F, Gremmels J, Schulz K, Messerschmidt K, 
            Mueller-Roeber B.
  TITLE     AssemblX: a user-friendly toolkit for rapid and reliable multi-gene 
            assemblies
  JOURNAL   Nucleic Acids Res. (2017) In press
  PUBMED    28130422
REFERENCE   2  (bases 1 to 5819)
  AUTHORS   Hochrein L, Machens F, Gremmels J, Schulz K, Messerschmidt K, 
            Mueller-Roeber B.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-NOV-2016) Department of Molecular Biology - Cell2Fab 
            research unit, University of Potsdam, Karl-Liebknecht-Str. 24-25, 
            Potsdam, Brandenburg 14476, Germany
REFERENCE   3  (bases 1 to 5819)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5819
                     /organism="Cloning vector pL0B_0-R"
                     /mol_type="other DNA"
                     /db_xref="taxon:1918743"
     CDS             complement(565..1368)
                     /codon_start=1
                     /product="URA3"
                     /label=URA3
                     /protein_id="AQK76578.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(1369..1584)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      1598..3069
                     /label=CEN/ARS origin
                     /note="CEN/ARS origin"
     rep_origin      2188..3068
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(3414..4208)
                     /codon_start=1
                     /product="neomycin phosphotransferase"
                     /label=neomycin phosphotransferase
                     /note="kanamycine resistance"
                     /protein_id="AQK76579.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHDDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(4209..4313)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    4382..4421
                     /label=8 cutter set: AscI, SbfI, SwaI, FseI, PmeI
                     /note="8 cutter set: AscI, SbfI, SwaI, FseI, PmeI"
     misc_feature    4422..4471
                     /label=homology region B0*
                     /note="homology region B0*"
     misc_feature    4478..5140
                     /label=ccdB cassette
                     /note="ccdB cassette"
     CDS             4807..5112
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     regulatory      5147..5551
                     /regulatory_class="promoter"
                     /note="LEU2 promoter"
     promoter        5147..5551
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     misc_feature    5552..5602
                     /label=homology region BR*
                     /note="homology region BR*"
     misc_feature    5606..5645
                     /label=8 cutter set: SwaI, AscI, PmeI, SbfI, FseI
                     /note="8 cutter set: SwaI, AscI, PmeI, SbfI, FseI"
     rep_origin      5655..455
                     /label=pBR322 origin
                     /note="pBR322 origin"
     rep_origin      5671..440
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"