LOCUS       Exported                6281 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Suicide vector pMQ200, complete sequence.
ACCESSION   FJ380063
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6281)
  AUTHORS   Shanks RM, Kadouri DE, MacEachran DP, OToole GA.
  TITLE     New yeast recombineering tools for bacteria
  JOURNAL   Plasmid 62 (2), 88-97 (2009)
  PUBMED    19477196
REFERENCE   2  (bases 1 to 6281)
  AUTHORS   Shanks RMQ., Kadouri DE, OToole GA.
  TITLE     Recombineering vectors for a wide range of Gram-negative and 
            -positive bacteria
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 6281)
  AUTHORS   Shanks RMQ., Kadouri DE, OToole GA.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-OCT-2008) Ophthalmololgy, University of Pittsburgh, 
            203 Lothrop St, Pittsburgh, PA 15213, USA
REFERENCE   4  (bases 1 to 6281)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6281
                     /organism="Suicide vector pMQ200"
                     /mol_type="other DNA"
                     /label=Cloning, suicide and expression vector
                     /note="Cloning, suicide and expression vector"
                     /db_xref="taxon:572233"
     CDS             1..804
                     /codon_start=1
                     /gene="ura3"
                     /product="Ura3"
                     /label=ura3
                     /protein_id="ACJ12803.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     gene            1..804
                     /gene="ura3"
                     /label=ura3
     oriT            8..1145
                     /label=conjugation transfer origin
                     /note="conjugation transfer origin"
     oriT            893..1001
                     /note="incP origin of transfer"
     promoter        complement(1192..1210)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      1418..1806
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     rep_origin      1478..1698
                     /label=oriR6K
                     /note="oriR6K"
     regulatory      complement(1808..2131)
                     /regulatory_class="terminator"
     terminator      1889..1975
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2141..2527)
                     /codon_start=1
                     /gene="lacZ alpha"
                     /product="beta-galactosidase alpha"
                     /label=lacZ alpha
                     /protein_id="ACJ12804.1"
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGETLADERRFSA"
     gene            complement(2141..2527)
                     /gene="lacZ alpha"
                     /label=lacZ alpha
     primer_bind     2311..2327
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2337..2355
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2364..2471
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2381..2397
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2431..2447)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2484..2502)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(2545..2829)
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     misc_feature    complement(2553..2580)
                     /label=Pbad
                     /note="Pbad"
     CDS             2805..3734
                     /codon_start=1
                     /gene="araC"
                     /product="AraC"
                     /label=araC
                     /protein_id="ACJ12805.1"
                     /translation="MQYGQLVSSLNGGSMKSMAEAQNDPLLPGYSFNAHLVAGLTPIEA
                     NGYLDFFIDRPLGMKGYILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPE
                     AREWYHQWVYFRPRAYWHEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGR
                     YSELLAINLLEQLLLRRMEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVC
                     LSPSRLSHLFRQQLGISVLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRV
                     FKKCTGASPSEFRAGCEEKVNDVAVKLS"
     gene            2805..3734
                     /gene="araC"
                     /label=araC
     CDS             2856..3734
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     CDS             4198..4992
                     /codon_start=1
                     /gene="nptII"
                     /product="NptII"
                     /label=nptII
                     /note="kanamycin resistance determinant"
                     /protein_id="ACJ12806.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            4198..4992
                     /gene="nptII"
                     /label=nptII
     promoter        complement(5148..5219)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5285..5801
                     /note="CEN6/ARSH4 for replication derived from 
                     Saccharomyces cerevisiae"
     misc_feature    5293..5796
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        6066..6281
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter