LOCUS       Exported                9606 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMQ97, complete sequence.
ACCESSION   DQ642043
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9606)
  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 9606)
  AUTHORS   Shanks RM, Caiazza NC, Hinsa SM, Toutain CM, OToole GA.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-MAY-2006) Microbiology and Immunology, Dartmouth 
            Medical School, Vail Building Room 505, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 9606)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9606
                     /organism="Shuttle vector pMQ97"
                     /mol_type="other DNA"
                     /label=yeast-bacteria shuttle vector
                     /note="yeast-bacteria shuttle vector"
                     /db_xref="taxon:390918"
     CDS             1..804
                     /codon_start=1
                     /gene="ura3"
                     /product="Ura3"
                     /label=ura3
                     /protein_id="ABG56599.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     gene            1..804
                     /gene="ura3"
                     /label=ura3
     misc_feature    886..1145
                     /label=oriT
                     /note="oriT"
     oriT            893..1001
                     /note="incP origin of transfer"
     rep_origin      1195..1642
                     /label=RK2 broad-host range origin of replication
                     /note="RK2 broad-host range origin of replication"
     rep_origin      1198..1728
                     /label=oriV
                     /note="incP origin of replication"
     CDS             2120..3268
                     /codon_start=1
                     /gene="trfA"
                     /product="TrfA"
                     /label=trfA
                     /note="RK2 replication protein"
                     /protein_id="ABG56600.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     gene            2120..3268
                     /gene="trfA"
                     /label=trfA
     regulatory      complement(3444..3767)
                     /regulatory_class="terminator"
                     /note="T1T2 transcriptional terminators from Escherichia 
                     coli rrnB"
     terminator      3525..3611
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3838..4554)
                     /codon_start=1
                     /gene="gfp"
                     /allele="mut3"
                     /product="Gfp"
                     /label=gfp
                     /protein_id="ABG56601.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     gene            complement(3838..4554)
                     /gene="gfp"
                     /allele="mut3"
                     /label=gfp
     regulatory      4562..4567
                     /regulatory_class="ribosome_binding_site"
                     /note="artificial RBS"
     promoter        complement(4612..4896)
                     /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(4620..4647)
                     /regulatory_class="promoter"
                     /note="PBAD"
     CDS             4872..5801
                     /codon_start=1
                     /gene="araC"
                     /product="AraC"
                     /label=araC
                     /protein_id="ABG56602.1"
                     /translation="MQYGQLVSSLNGGSMKSMAEAQNDPLLPGYSFNAHLVAGLTPIEA
                     NGYLDFFIDRPLGMKGYILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPE
                     AREWYHQWVYFRPRAYWHEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGR
                     YSELLAINLLEQLLLRRMEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVC
                     LSPSRLSHLFRQQLGISVLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRV
                     FKKCTGASPSEFRAGCEEKVNDVAVKLS"
     gene            4872..5801
                     /gene="araC"
                     /label=araC
     CDS             4923..5801
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     rep_origin      6133..6733
                     /label=ColE1 high copy pUC variant
                     /note="ColE1 high copy pUC variant"
     rep_origin      complement(6156..6744)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6953..7486)
                     /codon_start=1
                     /gene="aacC1"
                     /product="AacC1"
                     /label=aacC1
                     /note="gentamicin resistance determinant"
                     /protein_id="ABG56603.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            complement(6953..7486)
                     /gene="aacC1"
                     /label=aacC1
     promoter        complement(7675..7703)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      7780..9122
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      7780..8937
                     /label=2# high copy yeast origin
                     /note="2# high copy yeast origin"
     protein_bind    complement(8743..8790)
                     /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        9391..9606
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter