pCOMT-Kan vector (V008298)

LOCUS       V008298                14903 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V008298
VERSION     V008298
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 14903)
  AUTHORS   Staddon JH, Bryan EM, Manias DA, Dunny GM.
  TITLE     Conserved target for group II intron insertion in relaxase genes of
            conjugative elements of gram-positive bacteria
  JOURNAL   J. Bacteriol. 186 (8), 2393-2401 (2004)
   PUBMED   15060042
REFERENCE   2  (bases 1 to 14903)
  AUTHORS   Manias DA, Staddon JH, Klein JR, Dunny GM.
  TITLE     A new mobilizable shuttle plasmid based on pCOM9
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 14903)
  AUTHORS   Manias DA, Staddon JH, Klein JR, Dunny GM.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-MAY-2003) Microbiology, University of Minnesota, MMC
            196, 420 Delaware Street S.E., Minneapolis, MN 55455, USA
REFERENCE   4  (bases 1 to 14903)
  AUTHORS   Manias D, Dunny G.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-MAR-2007) Microbiology, Univ. Minnesota, 420 Delaware
            St., Minneapolis, MN 55455, USA
REFERENCE   5  (bases 1 to 14903)
  TITLE     Direct Submission
REFERENCE   6  (bases 1 to 14903)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J.
            Bacteriol."; date: "2004"; volume: "186"; issue: "8"; pages:
            "2393-2401"
            SGRef: number: 2; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
            (20-MAY-2003) Microbiology, University of Minnesota, MMC 196, 420
            Delaware Street S.E., Minneapolis, MN 55455, USA"
            SGRef: number: 4; type: "Journal Article"; journalName: "Submitted
            (05-MAR-2007) Microbiology, Univ. Minnesota, 420 Delaware St.,
            Minneapolis, MN 55455, USA"
            SGRef: number: 5; type: "Journal Article"
            On Mar 5, 2007 this sequence version replaced AY303238.1.
FEATURES             Location/Qualifiers
     source          1..14903
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(237..825)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     oriT            1069..1510
                     /label="pRS01 origin of transfer"
                     /note="pRS01 origin of transfer"
     CDS             2123..2890
                     /codon_start=1
                     /gene="AAD(9)"
                     /product="spectinomycin adenyltransferase"
                     /label="AAD(9)"
                     /note="spectinomycin resistance"
                     /protein_id="AAP60024.1"
                     /translation="MRRIYLNTYEQINKVKKILRKHLKNNLIGTYMFGSGVESGLKPNS
                     DLDFLVVVSEPLTDQSKEILIQKIRPISKKIGDKSNLRYIELTIIIQQEMVPWNHPPKQ
                     EFIYGEWLQELYEQGYIPQKELNSDLTIMLYQAKRKNKRIYGNYDLEELLPDIPFSDVR
                     RAIMDSSEELIDNYQDDETNSILTLCRMILTMDTGKIIPKDIAGNAVAESSPLEHRERI
                     LLAVRSYLGENIEWTNENVNLTINYLNNRLKKL"
     gene            2123..2890
                     /gene="AAD(9)"
                     /label="AAD(9)"
     terminator      5596..5623
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      5755..5798
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      complement(6094..6121)
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(6213..6299)
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     6773..6789
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    6808..6912
                     /label="ltrE, 3' fragment"
                     /note="ltrE, 3' fragment"
     CDS             6955..7311
                     /codon_start=1
                     /product="LtrF"
                     /function="unknown"
                     /label="LtrF"
                     /protein_id="AAP60025.1"
                     /translation="MKNKQNLKRPIQKIIRFSKEENDYLKDKIERSPFNNFQNYTRILL
                     LTGEVKIVDYTGLQKLNGEVNRIGNNINQIAKLAHQFKEISGEDINDLSQQLSEIQVMI
                     SERFKEELKQERLI"
     CDS             8364..10160
                     /gene="ltrA"
                     /label="Group II intron-encoded protein LtrA"
                     /note="Group II intron-encoded protein LtrA from
                     Lactococcus lactis subsp. cremoris (strain MG1363).
                     Accession#: P0A3U1"
     CDS             10410..11201
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     CDS             12891..14582
                     /codon_start=1
                     /product="LtrG*"
                     /label="LtrG*"
                     /protein_id="AAP60028.1"
                     /translation="MKLSFLHNYEVTTLAKAIYSITSWFPNRQFLGIVDKLNRDFIDNE
                     DKILGTIKIDSYTEFLNFYNQVLESYLPENPPEPFPIDKGNVKFYSNEEFHKIFIGNSS
                     EDIYEVSFIIESLIHDHESLKEIWYEILRYENLVISNLEQFKNSFIQEGNFECPPKDYF
                     YFIFNKFDEFRNDKLDEYFKEFKSVNVELYKFFSPVNKLPIFLPMMKECFIERVEEEIK
                     EEEFKESIWRSFYRILNCNFPLFQRAGGNIFYYLKLIHKDTKEEIDLENTIALLNENNL
                     IILESNISNIPEHIKVGIKNSLYRIAGLCQDGRVRVFEFRTECNIVFMGVDSEGISPNI
                     NKSIFLSRNEKIIFNASSIVGIINSAVDIKSIIDFLINYRKADDKMISFANIDAHFRTW
                     QLSNQVINEGASDLTIFYSTYESVHSNMDFFDRLQNLYPFELEGNFKNIHSWEIVEKED
                     TDLSLNSKVNSGSVDLFVVANKKIIYQEFDFILNDLDISDYEKISSFNEIILNALNKNK
                     KNILSNISGSILEINLVSQDILQKNSNHRYPIMNANYLESTCRSLA"
     misc_feature    12891..14551
                     /note="identical to pRS01 deposited under GenBank Accession
                     Number U50902"
     misc_feature    14552..14582
                     /label="from pDL281 vector"
                     /note="from pDL281 vector"
     primer_bind     complement(14588..14604)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    14612..14628
                     /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(14636..14666)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(14681..14702)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."