LOCUS       Exported                5973 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUC18T-miniTn7T-LAC-Zeo, complete sequence.
ACCESSION   MH290813
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5973)
  AUTHORS   Ducas-Mowchun K, DeSilva PM, Kumar A.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-MAY-2018) Department of Microbiology, University of 
            Manitoba, 45, Chancellor Circle, Buller Building (Room 412), 
            Winnipeg, MB R3T2N2, Canada
REFERENCE   2  (bases 1 to 5973)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: Geneious v. R11
            Sequencing Technology :: Illumina
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5973
                     /organism="Cloning vector pUC18T-miniTn7T-LAC-Zeo"
                     /mol_type="other DNA"
                     /db_xref="taxon:2282341"
     mobile_element  412..610
                     /mobile_element_type="transposon:Tn7R"
     primer_bind     615..631
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      659..753
                     /regulatory_class="terminator"
                     /note="lambda t0 terminator"
     terminator      659..753
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     regulatory      856..942
                     /regulatory_class="terminator"
                     /note="rrnB T1 terminator"
     terminator      856..942
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    988..1035
                     /label=FRT-R
                     /note="FRT-R"
     protein_bind    988..1035
                     /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)."
     CDS             complement(1145..1519)
                     /codon_start=1
                     /gene="bleoR"
                     /product="ZeoR"
                     /function="confers resistance to bleomycin, phleomycin, and
                     zeocin"
                     /label=bleoR
                     /protein_id="AXE73310.1"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     gene            complement(1145..1519)
                     /gene="bleoR"
                     /label=bleoR
     regulatory      complement(1538..1585)
                     /regulatory_class="promoter"
                     /note="EM7 promoter"
     promoter        complement(1538..1585)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     regulatory      complement(1764..1792)
                     /regulatory_class="promoter"
                     /note="Pc promoter"
     promoter        complement(1764..1792)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     misc_feature    1833..1880
                     /label=FRT-L
                     /note="FRT-L"
     protein_bind    1833..1880
                     /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)."
     CDS             complement(1923..3005)
                     /codon_start=1
                     /gene="lacI"
                     /product="LacI"
                     /label=lacI
                     /protein_id="AXE73309.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            complement(1923..3005)
                     /gene="lacI"
                     /label=lacI
     regulatory      complement(3006..3083)
                     /regulatory_class="promoter"
                     /note="lacIq promoter"
     promoter        complement(3006..3083)
                     /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."
     regulatory      3313..3341
                     /regulatory_class="promoter"
                     /note="tac promoter"
     promoter        3313..3341
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     regulatory      3349..3365
                     /regulatory_class="other"
                     /note="lac operator"
     protein_bind    3349..3365
                     /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)."
     mobile_element  3468..3633
                     /mobile_element_type="transposon:Tn7L"
     oriT            3792..3900
                     /note="incP origin of transfer"
     rep_origin      complement(4132..4720)
                     /direction=LEFT
     CDS             complement(4891..5751)
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase resistant protein"
                     /label=ampR
                     /note="AmpR"
                     /protein_id="AXE73308.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(4891..5751)
                     /gene="ampR"
                     /label=ampR
     regulatory      complement(5752..5856)
                     /regulatory_class="promoter"
                     /note="AmpR promoter"
     promoter        complement(5752..5856)
                     /gene="bla"
                     /label=AmpR promoter