LOCUS       Exported                6403 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUC18T-miniTn7T-LAC-Apr, complete sequence.
ACCESSION   MH290812
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6403)
  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 6403)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: Geneious v. R11
            Sequencing Technology :: Illumina
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6403
                     /organism="Cloning vector pUC18T-miniTn7T-LAC-Apr"
                     /mol_type="other DNA"
                     /db_xref="taxon:2282340"
     mobile_element  383..581
                     /mobile_element_type="transposon:Tn7R"
     primer_bind     586..602
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      630..724
                     /regulatory_class="terminator"
                     /note="lambda t0 terminator"
     terminator      630..724
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     regulatory      827..913
                     /regulatory_class="terminator"
                     /note="rrnB T1 terminator"
     terminator      827..913
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    959..1006
                     /label=FRT-R
                     /note="FRT-R"
     protein_bind    959..1006
                     /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(1121..1924)
                     /codon_start=1
                     /gene="aaC(3)-IV"
                     /product="apramycin resistance protein"
                     /label=aaC(3)-IV
                     /note="ApR"
                     /protein_id="AXE73306.1"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVISDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     gene            complement(1121..1924)
                     /gene="aaC(3)-IV"
                     /label=aaC(3)-IV
     CDS             complement(1124..1924)
                     /codon_start=1
                     /gene="aac(3)IV"
                     /product="aminoglycoside 3-N-acetyltransferase type IV     
                               "
                     /label=ApmR
                     /note="confers resistance to apramycin (Paget and Davies, 
                     1996)"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVISDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     regulatory      complement(2165..2193)
                     /regulatory_class="promoter"
                     /note="Pc promoter"
     promoter        complement(2165..2193)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     misc_feature    2234..2281
                     /label=FRT
                     /note="FRT"
     protein_bind    2234..2281
                     /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(2324..3406)
                     /codon_start=1
                     /gene="lacI"
                     /product="LacI"
                     /label=lacI
                     /protein_id="AXE73305.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            complement(2324..3406)
                     /gene="lacI"
                     /label=lacI
     regulatory      complement(3407..3484)
                     /regulatory_class="promoter"
                     /note="lacIq promoter"
     promoter        complement(3407..3484)
                     /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      3714..3742
                     /regulatory_class="promoter"
                     /note="tac promoter"
     promoter        3714..3742
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     regulatory      3750..3766
                     /regulatory_class="other"
                     /note="lac operator"
     protein_bind    3750..3766
                     /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)."
     misc_feature    3780..3860
                     /label=MCS
                     /note="MCS"
     mobile_element  3869..4034
                     /mobile_element_type="transposon:Tn7L"
     oriT            4193..4301
                     /note="incP origin of transfer"
     rep_origin      complement(4533..5121)
                     /direction=LEFT
     CDS             complement(5292..6152)
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase resistant protein"
                     /label=ampR
                     /note="AmpR"
                     /protein_id="AXE73307.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(5292..6152)
                     /gene="ampR"
                     /label=ampR
     regulatory      complement(6153..6257)
                     /regulatory_class="promoter"
                     /note="AmpR promoter"
     promoter        complement(6153..6257)
                     /gene="bla"
                     /label=AmpR promoter