LOCUS       Exported                5627 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pL0A_4-R, complete sequence.
ACCESSION   KY132035
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5627)
  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 5627)
  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 5627)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5627
                     /organism="Cloning vector pL0A_4-R"
                     /mol_type="other DNA"
                     /db_xref="taxon:1918741"
     CDS             complement(579..1382)
                     /codon_start=1
                     /product="URA3"
                     /label=URA3
                     /protein_id="AQK76574.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(1383..1598)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      1612..3083
                     /label=2micron origin
                     /note="2micron origin"
     rep_origin      2202..3082
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(3428..4222)
                     /codon_start=1
                     /product="neomycin phosphotransferase"
                     /label=neomycin phosphotransferase
                     /note="kanamycine resistance"
                     /protein_id="AQK76575.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHDDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(4223..4327)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    4396..4435
                     /label=8 cutter set: AscI, SbfI, SwaI, FseI, PmeI
                     /note="8 cutter set: AscI, SbfI, SwaI, FseI, PmeI"
     misc_feature    4436..4471
                     /label=homology region A4*
                     /note="homology region A4*"
     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..5373
                     /regulatory_class="promoter"
                     /note="URA3 promoter"
     promoter        5158..5373
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     misc_feature    5374..5424
                     /label=homology region AR*
                     /note="homology region AR*"
     misc_feature    5428..5467
                     /label=8 cutter set: SwaI, AscI, PmeI, SbfI, FseI
                     /note="8 cutter set: SwaI, AscI, PmeI, SbfI, FseI"
     rep_origin      5477..469
                     /label=pBR322 origin
                     /note="pBR322 origin"
     rep_origin      5493..454
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"