LOCUS       Exported                7489 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pX4, complete sequence.
ACCESSION   KC817032
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7489)
  AUTHORS   Kurosawa K, Wewetzer SJ, Sinskey AJ.
  TITLE     Engineering xylose metabolism in triacylglycerol-producing 
            Rhodococcus opacus for lignocellulosic fuel production
  JOURNAL   Biotechnol Biofuels 6 (1), 134 (2013)
  PUBMED    24041310
REFERENCE   2  (bases 1 to 7489)
  AUTHORS   Kurosawa K.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-MAR-2013) Biology, Massachusetts Institute of 
            Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 7489)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7489
                     /organism="Shuttle vector pX4"
                     /mol_type="other DNA"
                     /db_xref="taxon:1403748"
     CDS             complement(16..1248)
                     /codon_start=1
                     /gene="xylB"
                     /product="xylulose kinase"
                     /label=xylB
                     /protein_id="AGW51585.1"
                     /translation="MSAAEGPLVVGVDTSTQSTKALVVDAATGQVVASGQAPHTVTTGG
                     GRESDPRQWWDALCEALRQCGDAAHEAAAISVGGQQHGLVTLDERGEPVRPALLWNDVR
                     SAPQARRLVEELGGPKAWAERTGSVPGASFTASKWAWLAEHEPEAARATRAVRLPHDYL
                     TERLTGQAVTDRGDASGTGWWASGTERYDEDTLAHLGLDPALLPRVVRPGEVAGTVRDG
                     HGLPFSKGTLVASGTGDNAAAALGLGLLPGTPVLSLGTSGTVYAVSRHRPADPTGTVAG
                     FADARGDWLPLACTLNCTLAVDRVATLLGLDREAVAPGGSVTLLPFLDGERTPDLPNAS
                     GLLHGLRHDTTPGQLLQAAYDGAVHSLLGALDLVLDEDADRDSPLLLIGGGARGHAWQQ
                     TVRRLSGRPVQ"
     gene            complement(16..1248)
                     /gene="xylB"
                     /label=xylB
     protein_bind    1397..1413
                     /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(1421..1450)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             complement(1910..3076)
                     /codon_start=1
                     /gene="xylA"
                     /product="xylose isomerase"
                     /label=xylA
                     /protein_id="AGW51586.1"
                     /translation="MSFQPTPEDRFTFGLWTVGWQGRDPFGDATRPALDPVETVQRLAE
                     LGAYGVTFHDDDLIPFGSSDTERESHIKRFRQALDATGMTVPMATTNLFTHPVFKDGGF
                     TANDRDVRRYALRKTIRNIDLAAELGAKTYVAWGGREGAESGGAKDVRDALDRMKEAFD
                     LLGEYVIAQGYDLRFAIEPKPNEPRGDILLPTVGHALAFIERLERPELYGVNPEVGHEQ
                     MAGLNFPHGIAQALWAGKLFHIDLNGQSGIKYDQDLRFGAGDLRAAFWLVDLLETAGYE
                     GPRHFDFKPPRTEDFDGVWASAAGCMRNYLILKDRAAAFRADPEVQEALRAARLDQLAQ
                     ATAADGLDALLADRSAFEDFDVDAAAARGMAFEHLDQLAMDHLLGARG"
     gene            complement(1910..3076)
                     /gene="xylA"
                     /label=xylA
     protein_bind    3235..3251
                     /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(3259..3288)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             complement(3613..4404)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRQALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     oriT            5270..5379
                     /note="incP origin of transfer"
     CDS             5879..7330
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein"
                     /label=repA
                     /protein_id="AGW51587.1"
                     /translation="MSLQTHAQNAHTKTPTRVERDRGGLDNTPASASDRDLLRDHLGRD
                     VLHGSVTRDFKKAYRRNADGTNSPRMYRFETDALGRCEYAMLTTKQYAAVLVVDVDQVG
                     TAGGDPADLNPYVRDVVRSLITHSVGPAWVGINPTNGKAQFIWLIDPVYADRNGKSAQM
                     KLLAATTRVLGELLDHDPHFSHRFSRNPFYTGKAPTAYRWYRQHNRVMRLGDLIKQVRD
                     MAGHDQFNPTPRQQFSSGRELINAVKTRREEAQAFKALAQDVDAEIAGGLDQYDPELID
                     GVRVLWIVQGTAARDETAFRHALKTGHRLRQQGQRLTDAAIIDAYEHAYNVAHTHGGAG
                     RDNEMPPMRDRQTMARRVRGYVAQSKSETYSGSNAPGKATSSERKALATMGRRGGQKAA
                     QRWKTDPEGKYAQAQRSKLEKTHRKKKAQGRSTKSRISQMVNDQYFQTGTVPTWAEIGA
                     EVGVSRATVARHVAELKKSGDYPDV"
     gene            5879..7330
                     /gene="repA"
                     /label=repA