LOCUS       Exported                8636 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMQ2, complete sequence.
ACCESSION   EU546815
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8636)
  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 8636)
  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  JOURNAL   Unpublished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 8636)
  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 8636)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8636
                     /organism="Cloning vector pMQ2"
                     /mol_type="other DNA"
                     /label=shuttle vector
                     /note="shuttle vector"
                     /db_xref="taxon:515317"
     promoter        201..416
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             417..1220
                     /codon_start=1
                     /gene="URA3"
                     /product="Ura3"
                     /label=URA3
                     /note="uracil biosynthesis protein"
                     /protein_id="ACB45178.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     gene            417..1220
                     /gene="URA3"
                     /label=URA3
     rep_origin      complement(1351..1806)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      complement(1410..1806)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 ori"
     primer_bind     1951..1967
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1977..1995
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2021..2037
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      2274..2431
                     /label=palA lagging strand origin
                     /note="palA lagging strand origin"
     CDS             complement(2570..3220)
                     /codon_start=1
                     /gene="cat194"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat194
                     /note="confers resistance to chloramphenicol; derived from 
                     pC194"
                     /protein_id="ACB45179.1"
                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"
     gene            complement(2570..3220)
                     /gene="cat194"
                     /label=cat194
     CDS             complement(3389..3772)
                     /codon_start=1
                     /gene="rep"
                     /product="replication protein"
                     /label=rep
                     /protein_id="ACB45180.1"
                     /translation="MHLILSNMTFNKFLKYLISLFFLSILFHVITHKNNLVFTNYDNKK
                     SCFFPFLCMFFTSHLKRYINRYEKATFFALKTSHTNNLRVTSLAGNSYPYYQDKKEKDF
                     SLRSNPLKKHKRPHFLMWSLFFN"
     gene            complement(3389..3772)
                     /gene="rep"
                     /label=rep
     rep_origin      3427..3555
                     /label=oriV for pC194
                     /note="oriV for pC194"
     CDS             3677..4366
                     /codon_start=1
                     /gene="pre"
                     /product="Pre"
                     /label=pre
                     /note="pC194 OrfA"
                     /protein_id="ACB45181.1"
                     /translation="MCYNMEKYTEKKQRNQVFQKFIKRHIGENQMDLVEDCNTFLSFVA
                     DKTLEKQKLYKANSCKNRFCPVCAWRKARKDALGLSLMMQYIKQQEKKEFIFLTLTTPN
                     VMSDELENEIKRYNNSFRKLIKRKKVGSVIKGYVRKLEITYNKKRDDYNPHFHVLIAVN
                     KSYFTDKRYYISQQEWLDLWRDVTGISEITQVQVQKIRQNNNKELYEMAKYSGKDSDYL
                     INKSKSL"
     gene            3677..4366
                     /gene="pre"
                     /label=pre
     misc_feature    4613..4699
                     /label=palB
                     /note="palB"
     CDS             4713..4727
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     primer_bind     complement(4981..4997)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(5034..5052)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5073..5089)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5097..5113
                     /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(5115..5199)
                     /regulatory_class="promoter"
                     /note="Plac"
     promoter        complement(5121..5151)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      5452..6053
                     /label=ColE1 high copy variant
                     /note="ColE1 high copy variant"
     rep_origin      complement(5475..6063)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6234..7094)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="ACB45182.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(6234..7094)
                     /gene="bla"
                     /label=bla
     promoter        complement(7095..7199)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7226..8568
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      7226..8383
                     /note="2-micron high-copy origin derived from Saccharomyces
                     cerevisiae"
     protein_bind    complement(8189..8236)
                     /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)."