LOCUS       Exported                6654 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMQ113, complete sequence.
ACCESSION   EU546816
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6654)
  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 6654)
  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  JOURNAL   Unpublished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 6654)
  AUTHORS   Shanks RM, MacEachran DP, Powers SZ, Kadouri DE, OToole GA.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-MAR-2008) Ophthalmology, University of Pittsburgh, 203
            Lothrop St., Pittsburgh, PA 15213, USA
REFERENCE   4  (bases 1 to 6654)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6654
                     /organism="Cloning vector pMQ113"
                     /mol_type="other DNA"
                     /label=shuttle vector
                     /note="shuttle vector"
                     /db_xref="taxon:515307"
     oriT            53..877
                     /label=IncP
                     /note="IncP"
     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="ACB45183.1"
                     /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      complement(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."
     CDS             complement(2000..2011)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2145..2354
                     /codon_start=1
                     /product="OrfB"
                     /label=OrfB
                     /protein_id="ACB45184.1"
                     /translation="MGGKEANFASVLRPPIKCRVPIFVPKTLYPNWLKGLRGFSIANES
                     PTFSPTFFINLYLSSFIVVFMITK"
     CDS             2395..2556
                     /codon_start=1
                     /gene="repC"
                     /product="RepC"
                     /label=repC
                     /protein_id="ACB45185.1"
                     /translation="MVISESKKRVMISLTKEQDKKLTDMAKQKGFSKSAVAALAIEEYA
                     RKESEQKK"
     gene            2395..2556
                     /gene="repC"
                     /label=repC
     CDS             2623..3321
                     /codon_start=1
                     /gene="repA-ts"
                     /product="replication protein"
                     /label=repA-ts
                     /protein_id="ACB45186.1"
                     /translation="MAIKNTKARNFGFLLYPDSIPNDWKEKLESLGVSMAVSPLHDMDE
                     KKDKDTWNNSNIIQNGKHYKKPHYHVIYIARNPVTIESVRNKIKRKLGNSSVAHVEILD
                     YIKGSYEYLTHESKDAIAKNKHIYDKKDILNINDFDIDRYITLDESQKRELKNLLLDIV
                     DDYNLVNTKDLMAFIRLRGAEFGILNTNDVKDIVSTNSSAFRLWFEGNYQCGYRASYAK
                     VLDAETGEIK"
     gene            2623..3321
                     /gene="repA-ts"
                     /label=repA-ts
     CDS             3318..3467
                     /codon_start=1
                     /product="OrfD"
                     /label=OrfD
                     /protein_id="ACB45187.1"
                     /translation="MTNKEKELFAENEELKKEIKDLKERIERYREMEVELSTTIDLLRG
                     GIIE"
     regulatory      3466..3507
                     /regulatory_class="terminator"
                     /note="putative"
     CDS             complement(3725..4519)
                     /codon_start=1
                     /gene="aphA3"
                     /product="AphA3"
                     /label=aphA3
                     /note="confers resistance to kanamycin"
                     /protein_id="ACB45188.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            complement(3725..4519)
                     /gene="aphA3"
                     /label=aphA3
     CDS             complement(5099..5902)
                     /codon_start=1
                     /gene="URA3"
                     /product="Ura3"
                     /label=URA3
                     /note="uracil biosynthesis protein"
                     /protein_id="ACB45189.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     gene            complement(5099..5902)
                     /gene="URA3"
                     /label=URA3
     promoter        complement(5903..6118)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      6141..6654
                     /label=CEN6/ARSH4 derived from Saccharomyces cerevisiae
                     /note="CEN6/ARSH4 derived from Saccharomyces cerevisiae"
     misc_feature    6151..6654
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"