LOCUS       Exported                6339 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLOI2708, complete sequence.
ACCESSION   EU848570
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6339)
  AUTHORS   DeCrescenzo Henriksen E, Yomano LP, Gardner BC, Ingram LO, Doran 
            Peterson J.
  TITLE     Chromosomal Integration of the Klebsiella oxytoca casAB Genes for 
            Cellobiase Activity and the Erwinia chrysanthemi pelE Gene for 
            Pectate Lyase Activity in Escherichia coli KO11 for Fuel Ethanol 
            Production from Lignocellulosic Biomass
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6339)
  AUTHORS   DeCrescenzo Henriksen E, Yomano LP, Gardner BC, Ingram LO, Doran 
            Peterson J.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-JUN-2008) Microbiology & Cell Science, University of 
            Florida, Bldg 981, Museum Rd., Gainesville, FL 32611, USA
REFERENCE   3  (bases 1 to 6339)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6339
                     /organism="Cloning vector pLOI2708"
                     /mol_type="other DNA"
                     /db_xref="taxon:575198"
     CDS             1..1254
                     /codon_start=1
                     /gene="lacY"
                     /product="galactose permease"
                     /label=lacY
                     /note="derived from Escherichia coli"
                     /protein_id="ACK43810.1"
                     /translation="MYYLKNTNFWMFGLFFFFYFFIMGAYFPFFPIWLHDINHISKSDT
                     GIIFAAISLFSLLFQPLFGLLSDKLGLRKYLLWIITGMLVMFAPFFIFIFGPLLQYNIL
                     VGSIVGGIYLGFCFNAGAPAVEAFIEKVSRRSNFEFGRARMFGCVGWALCASIVGIMFT
                     INNQFVFWLGSGCALILAVLLFFAKTDAPSSATVANAVGANHSAFSLKLALELFRQPKL
                     WFLSLYVIGVSCTYDVFDQQFANFFTSFFATGEQGTRVFGYVTTMGELLNASIMFFAPL
                     IINRIGGKNALLLAGTIMSVRIIGSSFATSALEVVILKTLHMFEVPFLLVGCFKYITSQ
                     FEVRFSATIYLVCFCFFKQLAMIFMSVLAGNMYESIGFQGAYLVLGLVALGFTLISVFT
                     LSGPGPLSLLRRQVNEVA"
     gene            1..1254
                     /gene="lacY"
                     /label=lacY
     primer_bind     1316..1332
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1361..2259
                     /note="surrogate promoter from random Zymomonas mobilis CP4
                     genomic DNA fragment"
     CDS             2279..4144
                     /codon_start=1
                     /gene="casA"
                     /product="cellobiose-specific PTS permease"
                     /label=casA
                     /note="derived from Klebsiella oxytoca M5A1"
                     /protein_id="ACK43811.1"
                     /translation="MEYKALAQDILTRVGGRENIVSLVHCATRLRFKLKDSKKADAEGL
                     KANPGVIMVVESGGQFQVVIGNHVHDVWLAVRSEAGLTDDSEPVAVKGEKVSLIGQLID
                     IVSGIFTPFIGVLAASGILKGLLALAVVCGWLTTQQATYKIWFAASDALFFFFPLFLGY
                     TAGKKFGGNPYITMVIGGALTHPLMIQAFDASMAPGAATEYFLGIPVTFINYSSSVIPI
                     ILASWVSCWIEKRSNAILPSSMKNFFTPAICLAVVVPLTFLIIGPVATWLSQLLANGYQ
                     LIYQVAPWLAGAAMGALWQVCVIFGLHWGLIPLMINNLAVLGHDSMMPMLLPAVMGQVG
                     AALGIFLRTRDARQKVLAGSAVSAGIFGVTEPAIYGLNLPLRRPFIFGCVYWPIGGAMV
                     GFSDSHVSTYSFGFGNIITLAQMIPPEGIDATVWGGAAGMFASLIIACVLTLVAGLPRS
                     SAEQAAVVVAPASVNDILAPMTGSVLALDQVPDSTFASGLLGQGVAIIPSVGKVIAPFS
                     GEVASIFQTKHAIGLLSDSGIELLIHVGIDTVKLDGAPFTAHVKEGDKIKAGDLLLEFD
                     RQAILDAGYDLATPIIISNSDDFRTLDMVSASAVDAGQPLLSVSR"
     gene            2279..4144
                     /gene="casA"
                     /label=casA
     CDS             4162..5556
                     /codon_start=1
                     /gene="casB"
                     /product="phospho-cellobiase"
                     /label=casB
                     /note="derived from Klebsiella oxytoca M5A1; similar to 
                     glycohydrolase"
                     /protein_id="ACK43812.1"
                     /translation="MKTFPQAFLWGGATAANQVEGAYLEDGKGLTTSDVQPRGVFGDVV
                     ERVPGDSGIKDIAIDFYHRYPEDISLFAEMGFNCLRVSIAWARIFPHGDEAQPNEAGLA
                     FYDKLFDEMAKHNITPLVTLSHYEMPWALVKNYGGWGNRKVIGFFERYARTVFERYQAK
                     VKLWLTFNEINMSLHAPMTGVGLPADSSKAEVYQAIHHQLVASALAAKACHDIVPEGKI
                     GNMLLGGLMYPLSCKPDDIFETLQQNRSWQFFGDVQCRGAYPGYMLRYFRDNGINLDIT
                     DADRAALKETVDFISFSYYMTGCVTADEELNKKARGNILSMVPNPHLASSEWGWQIDPL
                     GLRTLLNVLWDRYQKPLFIVENGLGAKDKVEADGSINDDYRISYLNDHLVQVREAIEDG
                     VELMGYTSWGPIDLVSASKAEMSKRYGFIYVDRDDDGNGTLARSRKKSFWWYKEVIATN
                     GGSLKE"
     gene            4162..5556
                     /gene="casB"
                     /label=casB
     CDS             5734..6339
                     /codon_start=1
                     /gene="lacA"
                     /product="thiogalactoside acetyltransferase"
                     /label=lacA
                     /note="derived from Escherichia coli"
                     /protein_id="ACK43813.1"
                     /translation="MPMTERIRAGKLFTDMCEGLPEKRLRGKTLMYEFNHSHPSEVEKR
                     ESLIKEMFATVGENAWVEPPVYFSYGSNIHIGRNFYANFNLTIVDDYTVTIGDNVLIAP
                     NVTLSVTGHPVHHELRKNGEMYSFPITIGNNVWIGSHVVINPGVTIGDNSVIGAGSIVT
                     KDIPPNVVAAGVPCRVIREINDRDKHYYFKDYKVESSV"
     gene            5734..6339
                     /gene="lacA"
                     /label=lacA