LOCUS       Exported                8882 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pASC8057, complete sequence.
ACCESSION   KF781637
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8882)
  AUTHORS   Kurosawa K, Plassmeier J, Kalinowski J, Ruckert C, Sinskey AJ.
  TITLE     Engineering l-arabinose metabolism in triacylglycerol-producing 
            Rhodococcus opacus for lignocellulosic fuel production
  JOURNAL   Metab. Eng. (2015) In press
  PUBMED    25936337
REFERENCE   2  (bases 1 to 8882)
  AUTHORS   Kurosawa K.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-OCT-2013) Biology, Massachusetts Institute of 
            Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 8882)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8882
                     /organism="Shuttle vector pASC8057"
                     /mol_type="other DNA"
                     /db_xref="taxon:1551384"
     CDS             complement(73..1578)
                     /codon_start=1
                     /gene="araA"
                     /product="Arabinose isomerase"
                     /label=araA
                     /protein_id="AIS36086.1"
                     /translation="MTAPDQRPVIWFLTGSQHLYGADTLSQVAEQSRRIAGALRDARLP
                     ADVVWQPVLTDAAEIRRLCLAANADDSCVGVVAWMHTFSPAKMWIAGLDALRKPLLHLH
                     TQVNVDLPWSTIDMDFMNLNQAAHGDREFGHVQTRVGVARKTVAGHVGDPRTVARVAAW
                     TRAAIGAAELRTLRLARFGDNMRDVAVTEGDKVEAQLRFGVSVNTYGVADLAAAVDAVP
                     DTEVTALVEEYQDTYRVAPELRTGGERHTSLRYAARLEAGLRIFLAEGGFRAFTTNFED
                     LGALRQLPGLAVQRLMADGYGFGGEGDWKTATLLRALKTMAAQLPGGTSFMEDYTYHLR
                     PGEELILGAHMLEVCPSLTGTTPSCEIHPLAIGGREDPVRLVFDADPGPGLVVGLADLG
                     DRFRLVANEIEVVAPPEPLPRLPVARAVWRPRPDLRTSTESWLTAGAPHHTVLTAALGT
                     EELTDLADILGTELLTIDATTTVREFAREVRWNQAYHRLAQGL"
     gene            complement(73..1578)
                     /gene="araA"
                     /label=araA
     CDS             complement(1590..2282)
                     /codon_start=1
                     /gene="araD"
                     /product="Epimerase"
                     /label=araD
                     /protein_id="AIS36087.1"
                     /translation="MGTLPASAEEAVARLRREVGDIHQELVRYRLVVWTAGNVSARVPG
                     HDLLVIKPSGVPYEELSPRDMIVCDLDGTVVDGELSPSSDTAAHAYVYRHMPHVGGVVH
                     THSTYASAWAARGEAVPCVLTAMADEFGAEIPVGPFALIGDDSIGRGIVETLSGHRSPA
                     VLMRSHGVFTIGKDARAAVKAAVMCEDVARTVHIARQLGEPRPLAQADVDRLHDRYQNV
                     YGQPGAAR"
     gene            complement(1590..2282)
                     /gene="araD"
                     /label=araD
     CDS             complement(2445..4109)
                     /codon_start=1
                     /gene="araB"
                     /product="Ribulokinase"
                     /label=araB
                     /protein_id="AIS36088.1"
                     /translation="MITDLPAVTVGIDFGTLSGRAVVVTVADGTELGSAVHEYRHGVVD
                     AELPDGGPKLPPDWALQVPQDWRDVLRHAVPEALAAAGVRPEQVIGVGTDFTACTVLPV
                     TADGTPLCELPALRRRPHAYPKLWRHHAAQPQADRINRHAAREGRPWLARYGGRISSEW
                     EYAKALQLLEEDPEVYALTERWIEAADWIVWELTGAESRNLCTAGYKGIHQDGRRPDAD
                     FLAGLDPGFAGFTAKLDHPLSPLGAPAGALTARAAAATGLPPGIPVAVGNVDAHVTSAA
                     ARALDPGHLLAIMGTSTCHILNSDVLADVPGMCGVVRDGVVPGLWGYEAGQSGVGDIFA
                     WAARTAVPESYAAEARAMDVSVHELLTEKAAAQPVGAHGLLALDWHSGNRSVLVDHSLS
                     GLIVGLTLDTRPEDVYRALLEATAFGTRTVVEAFERAGVAVTEFTAAGGLLRNAFLMQL
                     YSDVLERPVNVIDSAQGPALGSAIHAAVAAGAHPDIRAASAAMGRIRRAAYRPDPDRAA
                     AYRELYREYQALHDHFGRGGSSVMHRLRELRRAATAG"
     gene            complement(2445..4109)
                     /gene="araB"
                     /label=araB
     regulatory      2606..2615
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    4633..4649
                     /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(4657..4686)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             complement(5011..5802)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVKLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     oriT            6668..6777
                     /note="incP origin of transfer"
     CDS             7272..8723
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein"
                     /label=repA
                     /protein_id="AIS36089.1"
                     /translation="MSLQTHAQNAHTKTPTRVERDRGGLDNTPASASDRDLLRDHLGRD
                     VLHGSVTRDFKKAYRRNADGTNSPRMYRFETDALGRCEYAMLTTKQYAAVLVVDVDQVG
                     TAGGDPADLNPYVRDVVRSLITHSVGPAWVGINPTNGKAQFIWLIDPVYADRNGKSAQM
                     KLLAATTRVLGELLDHDPHFSHRFSRNPFYTGKAPTAYRWYRQHNRVMRLGDLIKQVRD
                     MAGHDQFNPTPRQQFSSGRELINAVKTRREEAQAFKALAQDVDAEIAGGLDQYDPELID
                     GVRVLWIVQGTAARDETAFRHALKTGHRLRQQGQRLTDAAIIDAYEHAYNVAHTHGGAG
                     RDNEMPPMRDRQTMARRVRGYVAQSKSETYSGSNAPGKATSSERKALATMGRRGGQKAA
                     QRWKTDPEGKYAQAQRSKLEKTHRKKKAQGRSTKSRISQMVNDQYFQTGTVPTWAEIGA
                     EVGVSRATVARHVAELKKSGDYPDV"
     gene            7272..8723
                     /gene="repA"
                     /label=repA