LOCUS       Exported                7836 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMR361-K, complete sequence.
ACCESSION   KY767032
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7836)
  AUTHORS   Narayanan AM, Ramsey MM, Stacy A, Whiteley M.
  TITLE     Genomic resources for an oral pathogen
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7836)
  AUTHORS   Ramsey MM.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-MAR-2017) Department of Molecular Biosciences, The 
            University of Texas at Austin, 2506 Speedway, NMS 3.254, Austin, TX 
            78712, USA
REFERENCE   3  (bases 1 to 7836)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7836
                     /organism="Cloning vector pMR361-K"
                     /lab_host="Escherichia coli strain MFdpir"
                     /mol_type="other DNA"
                     /db_xref="taxon:1968886"
     oriT            35..144
                     /label=from incP
                     /note="from incP"
     CDS             177..548
                     /codon_start=1
                     /product="TraJ"
                     /label=TraJ
                     /note="oriT-recognizing protein"
                     /protein_id="ARI46003.1"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     CDS             604..1344
                     /codon_start=1
                     /product="TraI"
                     /label=TraI
                     /protein_id="ARI46004.1"
                     /translation="MRSIKKSDFAELVKYITDEQGKTERLGHVRVTNCEANTLPAVMAE
                     VMATQHGNTRSEADKTYHLLVSFRAGEKPDAETLRAIEDRICAGLGFAEHQRVSAVHHD
                     TDNLHIHIAINKIHPTRNTIHEPYRAYRALADLCATLERDYGLERDNHETRQRVSENRA
                     NDMERHAGVESLVGWILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFG
                     VGMVAGPVAGGLLGAISLLPRAFR"
     promoter        complement(1616..1720)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1918..2506
                     /label=high-copy-number ColE1/pMB1/pBR322/pUC
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC"
     protein_bind    2794..2815
                     /label=CAP (Escherichia coli catabolite activator prot
                     /bound_moiety="CAP (Escherichia coli catabolite activator 
                     protein)"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP"
     regulatory      2830..2860
                     /regulatory_class="promoter"
                     /note="from the Escherichia coli lac operon"
     promoter        2830..2860
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2868..2884
                     /label=lac repressor (encoded by lacI) binding site
                     /bound_moiety="lac repressor (encoded by lacI)"
                     /note="lac operator; the lac repressor binds to the lac 
                     operator to inhibit transcription in Escherichia coli; this
                     inhibition can be relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)"
     primer_bind     2892..2908
                     /note="M13 rev; common sequencing primer, one of multiple 
                     similar variants"
     CDS             2962..4008
                     /codon_start=1
                     /product="transposase"
                     /label=transposase
                     /protein_id="ARI46007.1"
                     /translation="MEKKEFRVLIKYCFLKGKNTVEAKTWLDNEFPDSAPGKSTIIDWY
                     AKFKRGEMSTEDGERSGRPKEVVTDENIKKIHKMILNDRKMKLIEIAEALKISKERVGH
                     IIHQYLDMRKLCAKWVPRELTFDQKQRRVDDSKRCLQLLTRNTPEFFRRYVTMDETWLH
                     HYTPESNRQSAEWTATGEPSPKRGKTQKSAGKVMASVFWDAHGIIFIDYLEKGKTINSD
                     YYMALLERLKVEIAAKRPHMKKKKVLFHQDNAPCHKSLRTMAKIHELGFELLPHPPYSP
                     DLAPSDFFLFSDLKRMLAGKKFGCNEEVIAETEAYFEAKPKEYYQNGIKKLEGRYNRCI
                     ALEGNYVE"
     mobile_element  4020..4098
                     /mobile_element_type="transposon:mariner"
                     /note="repeat end"
     regulatory      4104..4122
                     /regulatory_class="promoter"
                     /note="from bacteriophage T7 RNA polymerase"
     promoter        complement(4104..4122)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     4123..4139
                     /note="M13 rev; common sequencing primer, one of multiple 
                     similar variants"
     CDS             complement(4205..4999)
                     /codon_start=1
                     /product="aphA-3"
                     /label=aphA-3
                     /note="aminoglycoside phosphotransferase; confers 
                     resistance to kanamycin"
                     /protein_id="ARI46005.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     regulatory      5517..5535
                     /regulatory_class="promoter"
                     /note="from bacteriophage T7 RNA polymerase"
     promoter        5517..5535
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     mobile_element  5541..5640
                     /mobile_element_type="transposon:mariner"
                     /note="repeat end"
     CDS             6012..6686
                     /codon_start=1
                     /product="spectinomycin resistance cassette"
                     /label=spectinomycin resistance cassette
                     /note="SpcR"
                     /protein_id="ARI46006.1"
                     /translation="MFGSGVESGLKPNSDLDFLVVVSEPLTDQSKEILIQKIRPISKKI
                     GDKSNLRYIELTIIIQQEMVPWNHPPKQEFIYGEWLQELYEQGYIPQKELNSDLTIMLY
                     QAKRKNKRIYGNYDLEELLPDIPFSDVRRAIMDSSEELIDNYQDDETNSILTLCRMILT
                     MDTGKIIPKDIAGNAVAESSPLEHRERILLAVRSYLGENIEWTNENVNLTINYLNNRLK
                     KL"
     rep_origin      6871..7227
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"