LOCUS       Exported                8035 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMQ122, complete sequence.
ACCESSION   EU483152
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8035)
  AUTHORS   Shanks RMQ., MacEachran DP, OToole GA.
  TITLE     In vivo recombination vectors for modification and expression of 
            Gram-negative and Gram-positive bacterial genes
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8035)
  AUTHORS   Shanks RMQ., MacEachran DP, OToole GA.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-FEB-2008) Ophthalmololgy, University of Pittsburgh, 
            203 Lothrop st, Pittsburgh, PA 15213, USA
REFERENCE   3  (bases 1 to 8035)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8035
                     /organism="Shuttle vector pMQ122"
                     /mol_type="other DNA"
                     /db_xref="taxon:506624"
     CDS             1..804
                     /codon_start=1
                     /gene="ura3"
                     /product="Ura3"
                     /label=ura3
                     /note="uracil biosynthesis; URA3"
                     /protein_id="ACA49551.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     gene            1..804
                     /gene="ura3"
                     /label=ura3
     misc_feature    886..1145
                     /label=oriT from RP4
                     /note="oriT from RP4"
     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
                     /label=approximate OriV pRO1600 for pseudomonads
                     /note="approximate OriV pRO1600 for pseudomonads"
     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_id="ACA49552.1"
                     /translation="MASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     gene            1647..2480
                     /gene="rep"
                     /label=rep
     regulatory      complement(2598..2921)
                     /regulatory_class="terminator"
     terminator      2679..2765
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2992..3708)
                     /codon_start=1
                     /gene="GFP"
                     /allele="mut3"
                     /product="GFP"
                     /label=GFP
                     /protein_id="ACA49553.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     gene            complement(2992..3708)
                     /gene="GFP"
                     /allele="mut3"
                     /label=GFP
     regulatory      complement(3805..3825)
                     /regulatory_class="promoter"
                     /note="Ptac"
     protein_bind    3807..3823
                     /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(3831..3859)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     promoter        4089..4166
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             4290..5105
                     /codon_start=1
                     /gene="lacIQ"
                     /product="truncated LacIQ"
                     /label=lacIQ
                     /protein_id="ACA49554.1"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSR
                     ADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPA
                     LFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKY
                     LTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLR
                     VGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTTWTACCNSLRARR"
     gene            4290..5105
                     /gene="lacIQ"
                     /label=lacIQ
     rep_origin      5263..5863
                     /label=high copy ColE1 ori
                     /note="high copy ColE1 ori"
     rep_origin      complement(5286..5874)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6045..6905)
                     /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(6045..6563)
                     /codon_start=1
                     /gene="bla"
                     /product="AmpR determinant"
                     /label=bla
                     /protein_id="ACA49555.1"
                     /translation="MTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTR
                     LDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAE
                     IGASLIKHW"
     gene            complement(6045..6563)
                     /gene="bla"
                     /label=bla
     promoter        complement(6906..7010)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7039..7555
                     /note="CEN6/ARSH4 for replication in Saccharomyces 
                     cerevisiae"
     misc_feature    7047..7550
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        7820..8035
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter