LOCUS       Exported               14903 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle vector pCOMT-Kan, complete sequence.
ACCESSION   AY303238
VERSION     .
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  JOURNAL   Unpublished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)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     On Mar 5, 2007 this sequence version replaced AY303238.1.
FEATURES             Location/Qualifiers
     source          1..14903
                     /organism="Shuttle vector pCOMT-Kan"
                     /mol_type="other DNA"
                     /db_xref="taxon:233878"
     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)
     regulatory      5483..6624
                     /regulatory_class="terminator"
                     /note="rrnB T1T2 terminator"
     terminator      5596..5623
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      5715..5801
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6094..6121
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      6213..6299
                     /gene="Escherichia coli rrnB"
                     /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             join(7311..7790,11414..12625)
                     /codon_start=1
                     /product="LtrB"
                     /label=LtrB
                     /protein_id="AAP60026.2"
                     /translation="MVYTKHIIVHKLKHLRQAKDYVENAEKTLVNESNEDHLTNLFPYI
                     SNPDKTMSKQLVSGHGITNVYDAANEFIATKKLKALSKGTDFNFDPQTGKVRFNVESLE
                     KNNAVLGHHLIQSFSPDDNLTPEQIHEIGRQTILEFTGGEYEFVIATHVDREHIHNHII
                     FNSTNLYTGKQFDWKVIPKEKTKSGKAYDVTKNNFEKVSDKIASRYGAKIIEKSPGNSH
                     LKYTKWQTQSIYKSQIKQRLDYLLEMSSDIEDFKRKAQALNLSFDFSGKWTTYRLLDEP
                     QMKNTRGRNLDKNRPEKYNLESIIERLETNELSLTVDEVVERYEEKVDVVKQDFDYQVT
                     VEKGQIDHMTSKGFYLNVDFGIADRGQIFIGGYKVDQLENRDCVLYLKKNETFRLLSEK
                     EASFTKYLTGHDLAKQLGLYNGTVPLKKEPVISTINQLVDAINFLAEHGVTEGTQFNNM
                     ESQLMSALGEAEEKLYVIDNKIMELTKIAKLLIEKESDHSQAVINELENLGVGPSIKYQ
                     DIHQELQSEKMSRKILKNKFEQTVDEINTFNEIRVTTLEENKGKIL"
     intron          7791..11413
                     /note="Ll.ltrB group II intron"
     CDS             10410..11204
                     /codon_start=1
                     /product="APH-III"
                     /label=APH-III
                     /note="3,5aminoglycoside phosphotransferase; 
                     kanamycin-resistance; encoded on enterococcal plasmid pJH1"
                     /protein_id="AAP60027.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     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    14681..14702
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."