LOCUS       Exported                7978 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMQ61, complete sequence.
ACCESSION   DQ230324
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7978)
  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 7978)
  AUTHORS   Shanks RMQ., Caiazza NC, Hinsa SM, Toutain CM, OToole GA.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-SEP-2005) Microbiology and Immunology, Dartmouth 
            Medical School, Vail Building Room 505, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 7978)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7978
                     /organism="Shuttle vector pMQ61"
                     /mol_type="other DNA"
                     /db_xref="taxon:349297"
     CDS             1..804
                     /codon_start=1
                     /gene="ura3"
                     /product="Ura3"
                     /label=ura3
                     /note="derived from Saccharomyces cerevisiae"
                     /protein_id="ABB04082.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     gene            1..804
                     /gene="ura3"
                     /label=ura3
     misc_feature    complement(527..2504)
                     /label=loxP-kanMX-loxP cassette from pUG6
                     /note="loxP-kanMX-loxP cassette from pUG6"
     misc_feature    964..997
                     /label=loxP
                     /note="loxP"
     protein_bind    964..997
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      complement(998..1262)
                     /regulatory_class="terminator"
                     /note="TEF1 transcriptional terminator"
     gene            complement(1060..2416)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     terminator      1060..1257
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(1263..2072)
                     /codon_start=1
                     /gene="kanR"
                     /product="KanR"
                     /label=kanR
                     /protein_id="ABB04083.1"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(1263..2072)
                     /gene="kanR"
                     /label=kanR
     regulatory      complement(2073..2470)
                     /regulatory_class="promoter"
                     /note="TEF1 promoter"
     promoter        complement(2073..2416)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     misc_feature    complement(2471..2504)
                     /label=loxP
                     /note="loxP"
     protein_bind    2471..2504
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     rep_origin      2534..2843
                     /note="pRO1600 origin of replication; derived from pUCP20"
     rep_origin      2555..2906
                     /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             2920..3753
                     /codon_start=1
                     /gene="rep"
                     /product="Rep"
                     /label=rep
                     /note="replication protein from pRO1600"
                     /protein_id="ABB04084.1"
                     /translation="MASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     gene            2920..3753
                     /gene="rep"
                     /label=rep
     CDS             complement(3750..4130)
                     /codon_start=1
                     /gene="lacZ alpha"
                     /product="B-gal alpha peptide"
                     /label=lacZ alpha
                     /note="contains multicloning site driven by Plac for blue 
                     white screening"
                     /protein_id="ABB04085.1"
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLKGRPGRGGHGL"
     gene            complement(3750..4130)
                     /gene="lacZ alpha"
                     /label=lacZ alpha
     primer_bind     3914..3930
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3940..3958
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3967..4074
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     3984..4000
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4034..4050)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(4087..4105)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4126..4142)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4150..4166
                     /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)."
     promoter        complement(4174..4204)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      4505..5105
                     /note="ColE1 origin; high copy version from pRS426"
     rep_origin      complement(4528..5116)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5325..5858)
                     /codon_start=1
                     /gene="aacC1"
                     /product="AacC1"
                     /label=aacC1
                     /note="gentamicin resistance determinant"
                     /protein_id="ABB04086.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            complement(5325..5858)
                     /gene="aacC1"
                     /label=aacC1
     promoter        complement(6047..6075)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      6152..7494
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      6152..7309
                     /label=2-micron origin
                     /note="2-micron origin"
     protein_bind    complement(7115..7162)
                     /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        7763..7978
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter