LOCUS       Exported                6071 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMQ132, complete sequence.
ACCESSION   EU546820
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6071)
  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 6071)
  AUTHORS   Shanks RM, MacEachran DP, Powers SZ, Kadouri DE, OToole GA.
  TITLE     In vivo recombination vectors for modification and expression of 
            genes in Gram-negative and Gram-positive bacteria
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 6071)
  AUTHORS   Shanks RM, MacEachran DP, Powers SZ, Kadouri DE, OToole GA.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-MAR-2008) Ophthalmology, University of Pittsburgh, 203
            Lothrop St., Pittsburgh, PA 15213, USA
REFERENCE   4  (bases 1 to 6071)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6071
                     /organism="Cloning vector pMQ132"
                     /mol_type="other DNA"
                     /label=shuttle vector
                     /note="shuttle vector"
                     /db_xref="taxon:515315"
     oriT            53..877
                     /label=RP4
                     /note="RP4"
     oriT            399..508
                     /note="incP origin of transfer"
     regulatory      complement(929..1134)
                     /regulatory_class="terminator"
                     /note="rrnB T1T2"
     terminator      929..956
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      1048..1134
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1290..1574)
                     /codon_start=1
                     /gene="lacZalpha"
                     /product="beta-gal alpha peptide"
                     /label=lacZalpha
                     /protein_id="ACB45202.1"
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRAWLFWRMREDFQPDTD"
     gene            complement(1290..1574)
                     /gene="lacZalpha"
                     /label=lacZalpha
     primer_bind     1484..1500
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1504..1560
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1570..1586)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1594..1610
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     regulatory      1613..1696
                     /regulatory_class="promoter"
                     /note="Plac"
     promoter        complement(1618..1648)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1663..1684
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      1747..2603
                     /label=derived derived from pBBR1
                     /note="derived derived from pBBR1"
     rep_origin      1834..2603
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             2604..3266
                     /codon_start=1
                     /gene="rep"
                     /product="replication protein"
                     /label=rep
                     /note="derived from pBBR1"
                     /protein_id="ACB45203.1"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     gene            2604..3266
                     /gene="rep"
                     /label=rep
     CDS             complement(3547..4080)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=aacC1
                     /note="confers resistance to gentamycin"
                     /protein_id="ACB45204.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            complement(3547..4080)
                     /gene="aacC1"
                     /label=aacC1
     promoter        complement(4269..4297)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        complement(4315..4419)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(4516..5319)
                     /codon_start=1
                     /gene="URA3"
                     /product="Ura3"
                     /label=URA3
                     /note="uracil biosynthesis protein"
                     /protein_id="ACB45205.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     gene            complement(4516..5319)
                     /gene="URA3"
                     /label=URA3
     promoter        complement(5320..5535)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      5558..6071
                     /label=CEN6/ARSH4 derived from Saccharomyces cerevisiae
                     /note="CEN6/ARSH4 derived from Saccharomyces cerevisiae"
     misc_feature    5568..6071
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"