LOCUS       Exported                6004 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMQ56, complete sequence.
ACCESSION   DQ642033
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6004)
  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 6004)
  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 6004)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6004
                     /organism="Shuttle vector pMQ56"
                     /mol_type="other DNA"
                     /label=yeast-bacteria shuttle vector
                     /note="yeast-bacteria shuttle vector"
                     /db_xref="taxon:390908"
     CDS             1..804
                     /codon_start=1
                     /gene="ura3"
                     /product="Ura3"
                     /label=ura3
                     /protein_id="ABG56552.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     gene            1..804
                     /gene="ura3"
                     /label=ura3
     misc_structure  886..1145
                     /label=oriT
                     /note="oriT"
     oriT            893..1001
                     /note="incP origin of transfer"
     misc_feature    1198..1231
                     /label=loxP
                     /note="loxP"
     protein_bind    1198..1231
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     rep_origin      1261..1570
                     /note="approximate pRO1600 origin of replication from 
                     pUCP20"
     rep_origin      1282..1633
                     /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             1647..2480
                     /codon_start=1
                     /gene="rep"
                     /product="Rep"
                     /label=rep
                     /note="replication protein from cryptic Pseudomonas 
                     aeruginosa plasmid pRO1600"
                     /protein_id="ABG56553.1"
                     /translation="MASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     gene            1647..2480
                     /gene="rep"
                     /label=rep
     CDS             complement(2477..2857)
                     /codon_start=1
                     /gene="lacZalpha"
                     /product="LacZ-alpha"
                     /label=lacZalpha
                     /protein_id="ABG56554.1"
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLKGRPGRGGHGL"
     gene            complement(2477..2857)
                     /gene="lacZalpha"
                     /label=lacZalpha
     primer_bind     2641..2657
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2667..2685
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2694..2801
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2711..2727
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2761..2777)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2814..2832)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2853..2869)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2877..2893
                     /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(2901..2931)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      3232..3832
                     /label=ColE1 high copy pUC variant
                     /note="ColE1 high copy pUC variant"
     rep_origin      complement(3255..3843)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4014..4874)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             complement(4014..4679)
                     /codon_start=1
                     /gene="bla"
                     /product="Bla"
                     /label=bla
                     /protein_id="ABG56555.1"
                     /translation="MMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKH
                     LTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNE
                     AIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWF
                     IADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     gene            complement(4014..4679)
                     /gene="bla"
                     /label=bla
     promoter        complement(4875..4979)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5008..5524
                     /note="CEN6/ARSH4 for replication in Saccharomyces 
                     cerevisiae"
     misc_feature    5016..5519
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        5789..6004
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter