LOCUS       Exported                5743 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Staphylococcal allelic exchange vector pIMAY, complete sequence.
ACCESSION   JQ621981
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5743)
  AUTHORS   Monk IR, Shah IM, Xu M, Tan MW, Foster TJ.
  TITLE     Transforming the Untransformable: Application of Direct 
            Transformation To Manipulate Genetically Staphylococcus aureus and 
            Staphylococcus epidermidis
  JOURNAL   MBio 3 (2), e00277-11 (2012)
  PUBMED    22434850
REFERENCE   2  (bases 1 to 5743)
  AUTHORS   Monk IR.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-FEB-2012) Department of Microbiology, Trinity College 
            Dublin, Nassau St, Dublin, Dublin 2, Ireland
REFERENCE   3  (bases 1 to 5743)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5743
                     /organism="Staphylococcal allelic exchange vector pIMAY"
                     /mol_type="other DNA"
                     /db_xref="taxon:1167618"
     CDS             complement(157..168)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             302..511
                     /codon_start=1
                     /gene="repB"
                     /product="RepB"
                     /label=repB
                     /protein_id="AFG28167.1"
                     /translation="MGGKEANFASVLRPPIKCRVPIFVPKTLYPNWLKGLRGFSIANES
                     PTFSPTFFINLYLSSFIVVFMITK"
     gene            302..511
                     /gene="repB"
                     /label=repB
                     /note="derived from pVE6007"
     CDS             552..713
                     /codon_start=1
                     /gene="repC"
                     /product="RepC"
                     /label=repC
                     /protein_id="AFG28168.1"
                     /translation="MVISESKKRVMISLTKEQDKKLTDMAKQKGFSKSAVAALAIEEYA
                     RKESEQKK"
     gene            552..713
                     /gene="repC"
                     /label=repC
                     /note="derived from pVE6007"
     CDS             780..1478
                     /codon_start=1
                     /gene="repA(ts)"
                     /product="RepA(ts)"
                     /label=repA(ts)
                     /protein_id="AFG28172.1"
                     /translation="MAIKNTKARNFGFLLYPDSIPNDWKEKLESLGVSMAVSPLHDMDE
                     KKDKDTWNNSNIIQNGKHYKKPHYHVIYIARNPVTIESVRNKIKRKLGNSSVAHVEILD
                     YIKGSYEYLTHESKDAIAKNKHIYDKKDILNINDFDIDRYITLDESQKRELKNLLLDIV
                     DDYNLVNTKDLMAFIRLRGAEFGILNTNDVKDIVSTNSSAFRLWFEGNYQCGYRASYAK
                     VLDAETGEIK"
     gene            780..1478
                     /gene="repA(ts)"
                     /label=repA(ts)
                     /note="derived from pVE6007"
     CDS             1475..1624
                     /codon_start=1
                     /gene="repD"
                     /product="RepD"
                     /label=repD
                     /protein_id="AFG28169.1"
                     /translation="MTNKEKELFAENEELKKEIKDLKERIERYREMEVELSTTIDLLRG
                     GIIE"
     gene            1475..1624
                     /gene="repD"
                     /label=repD
                     /note="derived from pVE6007"
     CDS             complement(1824..2474)
                     /codon_start=1
                     /gene="cat"
                     /product="Cat"
                     /label=cat
                     /protein_id="AFG28170.1"
                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"
     gene            complement(1824..2474)
                     /gene="cat"
                     /label=cat
                     /note="derived from pIMC"
     regulatory      complement(2475..2674)
                     /regulatory_class="promoter"
     misc_feature    2675..2847
                     /label=multiple cloning site
                     /note="multiple cloning site"
     promoter        2675..2693
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    2706..2813
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2723..2739
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2773..2789)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2822..2840)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      3081..3992
     rep_origin      complement(3347..3892)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     oriT            4034..4143
                     /note="incP origin of transfer"
     ncRNA           4373..4933
                     /product="secY"
                     /label=antisense_RNA
                     /note="derived from pKOR1"
                     /ncRNA_class="antisense_RNA"
     regulatory      4934..5081
                     /regulatory_class="promoter"
     protein_bind    4957..4975
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    5042..5060
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     CDS             5082..5708
                     /codon_start=1
                     /gene="tetR"
                     /product="TetR"
                     /label=tetR
                     /protein_id="AFG28171.1"
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     gene            5082..5708
                     /gene="tetR"
                     /label=tetR
                     /note="derived from pKOR1"
     CDS             5085..5708
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"