LOCUS       Exported                7454 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle/allelic-replacement vector pMQ30, complete sequence.
ACCESSION   DQ230317
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7454)
  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 7454)
  AUTHORS   Shanks RMQ., Caiazza NC, Hinsa SM, Toutain CM, OToole GA.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-SEP-2005) Microbiology and Immunology, Dartmouth 
            Medical School, Vail Building Room 505, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 7454)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7454
                     /organism="Shuttle/allelic-replacement vector pMQ30"
                     /mol_type="other DNA"
                     /db_xref="taxon:349290"
     CDS             complement(209..1630)
                     /codon_start=1
                     /gene="sacB"
                     /product="SacB"
                     /label=sacB
                     /note="counter selectable marker for sucrose sensitivity 
                     from pEX18-Gm"
                     /protein_id="ABB04055.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     gene            complement(209..1630)
                     /gene="sacB"
                     /label=sacB
     promoter        complement(1631..2076)
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     oriT            2104..2928
                     /label=conjugal transfer origin from pEX18-Gm
                     /note="conjugal transfer origin from pEX18-Gm"
     oriT            2450..2559
                     /note="incP origin of transfer"
     terminator      2980..3007
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      3099..3185
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3341..3625)
                     /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="ABB04056.1"
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRAWLFWRMREDFQPDTD"
     gene            complement(3341..3625)
                     /gene="lacZ alpha"
                     /label=lacZ alpha
     primer_bind     3535..3551
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3555..3611
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(3621..3637)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3645..3661
                     /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(3669..3699)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3714..3735
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      4000..4600
                     /note="ColE1 origin; pUC derived; narrow host range; high 
                     copy number"
     rep_origin      complement(4023..4611)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4864..5397)
                     /codon_start=1
                     /gene="aacC1"
                     /product="AacC1"
                     /label=aacC1
                     /note="gentamicin resistance determinant"
                     /protein_id="ABB04057.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            complement(4864..5397)
                     /gene="aacC1"
                     /label=aacC1
     promoter        complement(5586..5614)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        complement(5632..5736)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(5833..6636)
                     /codon_start=1
                     /gene="ura3"
                     /product="Ura3"
                     /label=ura3
                     /protein_id="ABB04058.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     gene            complement(5833..6636)
                     /gene="ura3"
                     /label=ura3
     promoter        complement(6637..6852)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     misc_feature    6875..7388
                     /note="CEN6/ARSH4; Saccharomyces cerevisiae replicon from 
                     pYC2-CT"
     misc_feature    6885..7388
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"