LOCUS       Exported                6032 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMQ156, complete sequence.
ACCESSION   FJ380061
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6032)
  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 6032)
  AUTHORS   Shanks RMQ., Kadouri DE, OToole GA.
  TITLE     Recombineering vectors for a wide range of Gram-negative and 
            -positive bacteria
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 6032)
  AUTHORS   Shanks RMQ., Kadouri DE, OToole GA.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-OCT-2008) Ophthalmololgy, University of Pittsburgh, 
            203 Lothrop St, Pittsburgh, PA 15213, USA
REFERENCE   4  (bases 1 to 6032)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6032
                     /organism="Cloning vector pMQ156"
                     /mol_type="other DNA"
                     /db_xref="taxon:572232"
     CDS             1..804
                     /codon_start=1
                     /gene="ura3"
                     /product="Ura3"
                     /label=ura3
                     /protein_id="ACJ12795.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     gene            1..804
                     /gene="ura3"
                     /label=ura3
     oriT            886..1145
                     /label=conjugal transfer origin
                     /note="conjugal transfer origin"
     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 pRO1600 replication origin
                     /note="approximate pRO1600 replication origin"
     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="replication protein"
                     /label=rep
                     /note="derived from pRO1600"
                     /protein_id="ACJ12796.1"
                     /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(2921..3205)
                     /codon_start=1
                     /gene="lacZ alpha"
                     /product="B-gal alpha peptide"
                     /label=lacZ alpha
                     /protein_id="ACJ12797.1"
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRAWLFWRMREDFQPDTD"
     gene            complement(2921..3205)
                     /gene="lacZ alpha"
                     /label=lacZ alpha
     primer_bind     3115..3131
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3135..3191
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(3201..3217)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3225..3241
                     /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(3244..3327)
                     /regulatory_class="promoter"
                     /note="Plac promoter"
     promoter        complement(3249..3279)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3294..3315
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        complement(3366..3384)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      3594..3981
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     rep_origin      3653..3873
                     /label=oriR6K-gamma
                     /note="oriR6K-gamma"
     CDS             complement(4131..4664)
                     /codon_start=1
                     /gene="aacC1"
                     /product="AacC1"
                     /label=aacC1
                     /note="gentamicin resistance determinant"
                     /protein_id="ACJ12798.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            complement(4131..4664)
                     /gene="aacC1"
                     /label=aacC1
     promoter        complement(4853..4881)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        complement(4899..4970)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5036..5116
                     /label=CEN6 derived from Saccharomyces cerevisiae
                     /note="CEN6 derived from Saccharomyces cerevisiae"
     misc_feature    5044..5547
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     misc_feature    5117..5552
                     /label=ARSH4 derived from Saccharomyces cerevisiae
                     /note="ARSH4 derived from Saccharomyces cerevisiae"
     promoter        5817..6032
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter