LOCUS       Exported                6362 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pX2, complete sequence.
ACCESSION   KC817030
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6362)
  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 6362)
  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 6362)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6362
                     /organism="Shuttle vector pX2"
                     /mol_type="other DNA"
                     /db_xref="taxon:1403746"
     CDS             complement(22..1959)
                     /codon_start=1
                     /gene="xylB"
                     /product="xylulose kinase"
                     /label=xylB
                     /protein_id="AGW51580.1"
                     /translation="MSAAEGPLVVGVDTSTQSTKALVVDAATGQVVASGQAPHTVTTGG
                     GRESDPRQWWDALCEALRQCGDAAHEAAAISVGGQQHGLVTLDERGEPVRPALLWNDVR
                     SAPQARRLVEELGGPKAWAERTGSVPGASFTASKWAWLAEHEPEAARATRAVRLPHDYL
                     TERLTGQAVTDRGDASGTGWWASGTERYDEDTLAHLGLDPALLPRVVRPGEVAGTVRDG
                     HGLPFSKGTLVASGTGDNAAAALGLGLLPGTPVLSLGTSGTVYAVSRHRPADPTGTVAG
                     FADARGDWLPLACTLNCTLAVDRVATLLGLDREAVAPGGSVTLLPFLDGERTPDLPNAS
                     GLLHGLRHDTTPGQLLQAAYDGAVHSLLGALDLVLDEDADRDSPLLLIGGGARGHAWQQ
                     TVRRLSGRPVQILRLAEEEAKELREEARRAAEQHRELAESAAQQVRNDAESYATERKAK
                     AEDEGSRIVEKAKGEASQLRADAQKDAQSKREEADALFEETRAKAAQAAADFETNLAKR
                     REQSERDLASRQQKAEKRLAEIEHRAEQLRLEAEKLRTDAERRARQTVETAQRQAEDIV
                     ADANAKADRIRSESERELAALTNRRDSINAQLTNVREMLATLTGAAVAAAGTTGEDEPV
                     SRGVPAQQTR"
     gene            complement(22..1959)
                     /gene="xylB"
                     /label=xylB
     protein_bind    2108..2124
                     /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(2132..2161)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             complement(2486..3277)
                     /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            4143..4252
                     /note="incP origin of transfer"
     CDS             4752..6203
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein"
                     /label=repA
                     /protein_id="AGW51581.1"
                     /translation="MSLQTHAQNAHTKTPTRVERDRGGLDNTPASASDRDLLRDHLGRD
                     VLHGSVTRDFKKAYRRNADGTNSPRMYRFETDALGRCEYAMLTTKQYAAVLVVDVDQVG
                     TAGGDPADLNPYVRDVVRSLITHSVGPAWVGINPTNGKAQFIWLIDPVYADRNGKSAQM
                     KLLAATTRVLGELLDHDPHFSHRFSRNPFYTGKAPTAYRWYRQHNRVMRLGDLIKQVRD
                     MAGHDQFNPTPRQQFSSGRELINAVKTRREEAQAFKALAQDVDAEIAGGLDQYDPELID
                     GVRVLWIVQGTAARDETAFRHALKTGHRLRQQGQRLTDAAIIDAYEHAYNVAHTHGGAG
                     RDNEMPPMRDRQTMARRVRGYVAQSKSETYSGSNAPGKATSSERKALATMGRRGGQKAA
                     QRWKTDPEGKYAQAQRSKLEKTHRKKKAQGRSTKSRISQMVNDQYFQTGTVPTWAEIGA
                     EVGVSRATVARHVAELKKSGDYPDV"
     gene            4752..6203
                     /gene="repA"
                     /label=repA
ORIGIN
        1 gaattctaga ctagtggaac cctatcgggt ctgctgcgcc ggaaccccac gagaaaccgg
       61 ctcgtcctca cccgtcgtgc cggcggcggc caccgccgcg ccggtcagcg tcgccagcat
      121 ctcgcgcacg ttcgtcagct gggcgttgat gctgtcgcgg cggttggtga gggccgccag
      181 ctcgcgctcg gactcggaac ggatgcggtc cgccttggcg ttggcgtcgg ccacgatgtc
      241 ctcggcctgg cgctgcgcgg tctccaccgt ctggcgggcg cggcgctccg cgtcggtgcg
      301 cagcttctcc gcctccaggc gcagctgctc ggcgcggtgc tcgatctccg cgagccgctt
      361 ctccgccttc tgctgacgcg aggccaggtc gcgctcggac tgctcgcggc gcttggccag
      421 gttcgtctcg aagtcggcgg cggcctgcgc ggccttggcg cgggtctcct cgaagagggc
      481 gtccgcctcc tcacgcttgg actgcgcgtc cttctgcgcg tcggcacgca gctgagaggc
      541 ctcgcccttg gccttctcga cgatccggga gccctcgtcc tccgccttcg ccttgcgctc
      601 ggtggcgtag gactcggcgt cgttacgcac ctgctgggcg gccgactcgg ccagctcgcg
      661 gtgctgttcg gccgcgcggc gggcctcctc acgcagctcc ttggcctctt cctcggccag
      721 gcgcaggatc tggaccggcc gtccggacag ccgtcgcacg gtctgctgcc aggcgtggcc
      781 ccgcgcgccg ccgccgatca gcagcagcgg gctgtcgcgg tccgcgtcct cgtcgaggac
      841 caggtcgagc gcgccgagca gggagtgcac cgcaccgtcg tacgccgcct ggagcagctg
      901 gcccggggtc gtgtcgtggc gcagcccgtg cagcagaccg gaggcgttcg gcaggtcggg
      961 ggtgcgttcg ccgtccagga acggcaggag cgtcacggag ccgccgggcg cgacggcctc
     1021 gcggtccagg cccagcagag tcgcgacgcg gtcgacggcg agcgtgcagt tcagggtgca
     1081 ggccagcggc agccagtccc cgcgcgcgtc ggcgaagccc gccaccgtac cggtgggatc
     1141 ggccgggcgg tgccgggaca cggcgtacac cgtgccggag gtcccgaggc tcagtaccgg
     1201 ggtgccgggc agcagcccga ggcccagcgc ggccgccgcg ttgtcaccgg tgcccgaggc
     1261 caccaacgtg cccttggaga aaggcagacc gtgtccgtcg cgtacggtcc cggccacctc
     1321 acccggacgc accacccggg gcagcagcgc cgggtcgagc ccgaggtgcg ccagagtgtc
     1381 ctcgtcgtac cgctccgtgc cggacgccca ccagccggtg ccggaggcgt caccgcgatc
     1441 ggtgacggcc tgcccggtca ggcgctcggt gaggtagtcg tgggggagcc gtacggccct
     1501 ggtcgcgcgg gccgcctccg gttcgtgctc ggcgagccaa gcccacttgc tcgccgtgaa
     1561 cgaggccccg ggcacgctgc cggtgcgttc ggcccaagcc ttcgggccgc ccagttcctc
     1621 caccagccga cgggcctgcg gcgcggagcg gacatcgttc cagagcagcg ccggccgcac
     1681 cggctcaccc cgctcgtcca gcgtgaccag gccgtgctgc tgtccgccga ccgagatcgc
     1741 ggccgcctcg tgcgccgcgt caccgcactg gcgcagcgcc tcgcacaggg cgtcccacca
     1801 ctggcgcgga tcgctctccc ggcccccgcc ggtggtgacg gtgtgcggcg cctggccgct
     1861 cgccaccacc tgtccggtgg ccgcgtcgac gaccagggcc ttcgtggact gggtggacgt
     1921 gtccacgccg acgacgagcg gaccctccgc tgctgacatg gggttctccc tcttccgcgg
     1981 ctcagcggga tgtgctgtgg ggcgctcccg aggacacccc cgagggcgcc cgtggaagac
     2041 atctgtcgaa gacaccttgt ggtcgaagac accggatcca attccatggt ctgtttcctg
     2101 tgtgaaattg ttatccgctc acaattccac acattatacg agccggatga ttaattgtca
     2161 acagctcatt tcagaatatt tgccagaacc gttatgatgt cggcgcaaaa aacattatcc
     2221 agaacgggag tgcgccttga gcgacacgaa ttatgcagtg atttacgacc tgcacagcca
     2281 taccacagct tccgatggct gcctgacgcc agaagcattg gtgcaccgtg cagtcgatga
     2341 taagctgtca aaccagatca attcgcgcta acttacatta attgcgttgc gctcactgcc
     2401 cgctttccag tcgggaaacc tgtcgtgcca gctgaaacgc ggttacaatt cgcccttcca
     2461 attcgccctt gcaacgcgtt tcgaattatt tgccgactac cttggtgatc tcgcctttca
     2521 cgtagtggac aaattcttcc aactgatctg cgcgcgaggc caagcgatct tcttcttgtc
     2581 caagataagc ctgtctagct tcaagtatga cgggctgata ctgggccggc aggcgctcca
     2641 ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc ggttactgcg ctgtaccaaa
     2701 tgcgggacaa cgtaagcact acatttcgct catcgccagc ccagtcgggc ggcgagttcc
     2761 atagcgttaa ggtttcattt agcgcctcaa atagatcctg ttcaggaacc ggatcaaaga
     2821 gttcctccgc cgctggacct accaaggcaa cgctatgttc tcttgctttt gtcagcaaga
     2881 tagccagatc aatgtcgatc gtggctggct cgaagatacc tgcaagaatg tcattgcgct
     2941 gccattctcc aaattgcagt tcgcgcttag ctggataacg ccacggaatg atgtcgtcgt
     3001 gcacaacaat ggtgacttct acagcgcgga gaatctcgct ctctccaggg gaagccgaag
     3061 tttccaaaag gtcgttgatc aaagcttgcc gcgttgtttc atcaagcctt acggtcaccg
     3121 taaccagcaa atcaatatca ctgtgtggct tcaggccgcc atccactgcg gagccgtaca
     3181 aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat gacgccaact acctctgata
     3241 gttgagtcga tacttcggcg atcaccgctt ccctcatggg gggcctcctc cgctaaccgt
     3301 ggatctagta cttcattggc agctaatccg gaatcgcact tacggccaat gcttcgtttc
     3361 gtatcacaca ccccaaagcc ttctgctttg aatgctgccc ttcttcaggg cttaattttt
     3421 aagagcgtca ccttcatggt ggtcagtgcg tcctgctgat gtgctcagta tcaccgccag
     3481 tggtatttat gtcgctcagc acgcgttgca agggcgaatt gatccttttt gtccggtgtt
     3541 gggttgaagg tgaagccggt cggggccgca gcgggggccg gcttttcagc cttgcccccc
     3601 tgcttcggcc gccgtggctc cggcgtcttg ggtgccggcg cgggttccgc agccttggcc
     3661 tgcggtgcgg gcacatcggc gggcttggcc ttgatgtgcc gcctggcgtg cgagcggaac
     3721 gtctcgtagg agaacttgac cttccccgtt tcccgcatgt gctcccaaat ggtgacgagc
     3781 gcatagccgg acgctaacgc cgcctcgaca tccgccctca ccgccaggaa cgcaaccgca
     3841 gcctcatcac gccggcgctt cttggccgcg cgggattcaa cccactcggc cagctcgtcg
     3901 gtgtagctct ttggcatcgt ctctcgcctg tcccctcagt tcagtaattt cctgcatttg
     3961 cctgtttcca gtcggtagat attccacaaa acagcaggga agcagcgctt ttccgctgca
     4021 taaccctgct tcggggtcat tatagcgatt ttttcggtat atccatcctt tttcgcacga
     4081 tatacaggat tttgccaaag ggttcgtgta gactttcctt ggtgtatcca acggcgtcag
     4141 ccgggcagga taggtgaagt aggcccaccc gcgagcgggt gttccttctt cactgtccct
     4201 tattcgcacc tggcggtgct caacgggaat cctgctctgc gaggctggcc ggctaccgcc
     4261 ggcgtaacag atgagggcaa gcggatggct gatgaaacca agccaaccag gaagggcagc
     4321 ccacctatca aggtgtactg ccttccagac gaacgaagag cgattgagga aaaggcggcg
     4381 gcggccggca tgagcctgtc ggcctacctg ctggccgtcg gccagggcta caaaatcacg
     4441 ggcgtcgtgg actatgagca cgtccgcgag ctggcccgca tcaatggcga cctgggccgc
     4501 ctgggcggcc tgctgaaact ctggctcacc gacgacccgc gcacggcgcg gttcggtgat
     4561 gccacgatct ctagagcaag ggcgaattgt aaccgcaggt aaaaggcata tttttcgcgt
     4621 gtcatggcta gtaaataaca ccggtgtcat ttagagtcag ggaaagacaa tgaaaaacga
     4681 agaaagccac cgggcggcaa cccgatgact ttcgcttatc acccagcaca cacctgggag
     4741 aaatcacggt catgagttta cagactcatg cgcagaatgc gcacactaaa acacctaccc
     4801 gcgtcgagcg cgaccgtggt ggactggaca acaccccagc atctgccagt gaccgcgacc
     4861 ttttacgcga tcatctaggc cgcgatgtac tccacggttc agtcacacga gactttaaaa
     4921 aggcctatcg acgcaacgct gacggcacga actcgccgcg tatgtatcgc ttcgagactg
     4981 atgctttagg acggtgcgag tacgccatgc tcaccaccaa gcagtacgcc gccgtcctgg
     5041 tcgtagacgt tgaccaagta ggtaccgcag gcggtgaccc cgcagactta aacccgtacg
     5101 tccgcgacgt ggtgcgctca ctgattactc atagcgtcgg gccagcctgg gtgggtatta
     5161 acccaactaa cggcaaagcc cagttcatat ggcttattga ccctgtctac gctgaccgta
     5221 acggtaaatc tgcgcagatg aagcttcttg cagcaaccac gcgtgtgctg ggtgagcttt
     5281 tagaccatga cccgcacttt tcccaccgct ttagccgcaa cccgttctac acaggcaaag
     5341 cccctaccgc ttatcgttgg tataggcagc acaaccgggt gatgcgcctt ggagacttga
     5401 taaagcaggt aagggatatg gcaggacacg accagttcaa ccccacccca cgccagcaat
     5461 tcagctctgg ccgcgaactt atcaacgcgg tcaagacccg ccgtgaagaa gcccaagcat
     5521 tcaaagcact cgcccaggac gtagacgcgg aaatcgccgg tggtctcgac cagtatgacc
     5581 cggaacttat cgacggtgtg cgtgtgctct ggattgtcca aggaaccgca gcacgcgacg
     5641 aaacagcctt tagacatgcg cttaagactg gccaccgctt gcgccagcaa ggccaacgcc
     5701 tgacagacgc agcaatcatc gacgcctatg agcacgccta caacgtcgca cacacccacg
     5761 gcggtgcagg ccgcgacaac gagatgccac ccatgcgcga ccgccaaacc atggcaaggc
     5821 gcgtgcgcgg gtatgtcgcc caatccaaga gcgagaccta cagcggctct aacgcaccag
     5881 gtaaagccac cagcagcgag cggaaagcct tggccacgat gggacgcaga ggcggacaaa
     5941 aagccgcaca acgctggaaa acagaccccg agggcaaata tgcgcaagca caaaggtcga
     6001 agcttgaaaa gacgcaccgt aagaaaaagg ctcaaggacg atctacgaag tcccgtatta
     6061 gccaaatggt gaacgatcag tatttccaga cagggacagt tcccacgtgg gctgaaatag
     6121 gggcagaggt aggagtctct cgcgccacgg ttgctaggca tgtcgcggag ctaaagaaga
     6181 gcggtgacta tccggacgtt taaggggtct cataccgtaa gcaatatacg gttcccctgc
     6241 cgttaggcag ttagataaaa cctcacttga agaaaacctt gaggggcagg gcagcttata
     6301 tgcttcaaag catgacttcc tctgttctcc tagacctcgc aaccctccgc cataacctca
     6361 cc
//
