LOCUS       Exported                9312 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMQ71, complete sequence.
ACCESSION   DQ642036
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9312)
  AUTHORS   Shanks RM, Caiazza NC, Hinsa SM, Toutain CM, OToole GA.
  TITLE     Saccharomyces cerevisiae-based molecular tool kit for manipulation 
            of genes from gram-negative bacteria
  JOURNAL   Appl. Environ. Microbiol. 72 (7), 5027-5036 (2006)
  PUBMED    16820502
REFERENCE   2  (bases 1 to 9312)
  AUTHORS   Shanks RM, Caiazza NC, Hinsa SM, Toutain CM, OToole GA.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-MAY-2006) Microbiology and Immunology, Dartmouth 
            Medical School, Vail Building Room 505, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 9312)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9312
                     /organism="Shuttle vector pMQ71"
                     /mol_type="other DNA"
                     /label=yeast-bacteria shuttle vector
                     /note="yeast-bacteria shuttle vector"
                     /db_xref="taxon:390911"
     CDS             87..890
                     /codon_start=1
                     /gene="ura3"
                     /product="Ura3"
                     /label=ura3
                     /protein_id="ABG56564.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     gene            87..890
                     /gene="ura3"
                     /label=ura3
     misc_feature    613..2590
                     /label=loxP-kanMX-loxP from pUG6
                     /note="loxP-kanMX-loxP from pUG6"
     regulatory      complement(998..1262)
                     /regulatory_class="terminator"
                     /note="TEF1 terminator"
     protein_bind    1050..1083
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_recomb     1136..1169
                     /label=loxP
                     /note="loxP"
     gene            complement(1146..2502)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     terminator      1146..1343
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(1349..2158)
                     /codon_start=1
                     /gene="kanR"
                     /product="KanR"
                     /label=kanR
                     /note="confers resistance to kanamycin in Gram-negatives 
                     and G418 sulfonate in yeast"
                     /protein_id="ABG56565.1"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(1349..2158)
                     /gene="kanR"
                     /label=kanR
     regulatory      complement(2159..2556)
                     /regulatory_class="promoter"
                     /note="TEF1 promoter"
     promoter        complement(2159..2502)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     misc_recomb     2557..2590
                     /label=loxP
                     /note="loxP"
     protein_bind    2557..2590
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     rep_origin      2620..2929
                     /note="approximate pRO1600 origin of replication from 
                     pUCP20"
     rep_origin      2641..2992
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from 
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"
     CDS             3006..3839
                     /codon_start=1
                     /gene="rep"
                     /product="Rep"
                     /label=rep
                     /protein_id="ABG56566.1"
                     /translation="MASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     gene            3006..3839
                     /gene="rep"
                     /label=rep
     regulatory      complement(3957..4280)
                     /regulatory_class="terminator"
                     /note="T1T2 transcriptional terminators from Escherichia 
                     coli rrnB"
     terminator      4038..4124
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    4327..4383
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     promoter        complement(4404..4688)
                     /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)"
     regulatory      complement(4412)
                     /regulatory_class="promoter"
                     /note="PBAD"
     CDS             4664..5593
                     /codon_start=1
                     /gene="araC"
                     /product="AraC"
                     /label=araC
                     /protein_id="ABG56567.1"
                     /translation="MQYGQLVSSLNGGSMKSMAEAQNDPLLPGYSFNAHLVAGLTPIEA
                     NGYLDFFIDRPLGMKGYILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPE
                     AREWYHQWVYFRPRAYWHEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGR
                     YSELLAINLLEQLLLRRMEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVC
                     LSPSRLSHLFRQQLGISVLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRV
                     FKKCTGASPSEFRAGCEEKVNDVAVKLS"
     gene            4664..5593
                     /gene="araC"
                     /label=araC
     CDS             4715..5593
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     rep_origin      5925..6525
                     /label=ColE1 high copy pUC variant
                     /note="ColE1 high copy pUC variant"
     rep_origin      complement(5948..6536)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6745..7278)
                     /codon_start=1
                     /gene="aacC1"
                     /product="AacC1"
                     /label=aacC1
                     /protein_id="ABG56568.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            complement(6745..7278)
                     /gene="aacC1"
                     /label=aacC1
     promoter        complement(7467..7495)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      7572..8914
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      7572..8729
                     /label=2# high copy yeast origin
                     /note="2# high copy yeast origin"
     protein_bind    complement(8535..8582)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        9183..86
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter