LOCUS       Exported                4177 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pX1918 with xylE cassette, complete sequence.
ACCESSION   U03992
VERSION     .
KEYWORDS    beta-lactamase; catechol-2,3-dioxygenase.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4177)
  AUTHORS   Schweizer HP.
  TITLE     Two plasmids, X1918 and Z1918, for easy recovery of the xylE and 
            lacZ reporter genes
  JOURNAL   Gene 134 (1), 89-91 (1993)
  PUBMED    8244036
REFERENCE   2  (bases 1 to 4177)
  AUTHORS   Harayama S, Rekik M, Bairoch A, Neidle EL, Ornston LN.
  TITLE     Potential DNA slippage structures acquired during evolutionary 
            divergence of Acinetobacter calcoaceticus chromosomal benABC and 
            Pseudomonas putida TOL pWW0 plasmid xylXYZ, genes encoding benzoate 
            dioxygenases
  JOURNAL   J. Bacteriol. 173 (23), 7540-7548 (1991)
  PUBMED    1938949
REFERENCE   3  (bases 1 to 4177)
  AUTHORS   Deretic V, Gill JF, Chakrabarty AM.
  TITLE     Pseudomonas aeruginosa infection in cystic fibrosis: nucleotide 
            sequence and transcriptional regulation of the algD gene
  JOURNAL   Nucleic Acids Res. 15 (11), 4567-4581 (1987)
  PUBMED    3108855
REFERENCE   4  (bases 1 to 4177)
  AUTHORS   Konyecsni WM, Deretic V.
  TITLE     Broad-host-range plasmid and M13 bacteriophage-derived vectors for 
            promoter analysis in Escherichia coli and Pseudomonas aeruginosa
  JOURNAL   Gene 74 (2), 375-386 (1988)
  PUBMED    3149945
REFERENCE   5  (bases 1 to 4177)
  AUTHORS   Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-DEC-1993) Herbert P. Schweizer, Microbiology and 
            Infectious Diseases, University of Calgary, 3330 Hospital Drive 
            N.W., Calgary, Alberta, T2N 4N1, Canada
REFERENCE   6  (bases 1 to 4177)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     On Dec 16, 1993 this sequence version replaced gi:434649.
FEATURES             Location/Qualifiers
     source          1..4177
                     /organism="Cloning vector pX1918"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /note="nucleotides 1-449, 1893-4177 from vector pUC1918; 
                     450-1892 from phage mVDX18"
                     /db_xref="taxon:31866"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    399..455
                     /standard_name="polylinker"
                     /label=polylinker
     misc_feature    450..1892
                     /label=insert from mVDX18
                     /note="insert from mVDX18"
                     /citation="[4]"
     misc_feature    456..544
                     /label=algD DNA from Pseudomonas aeruginosa
                     /note="algD DNA from Pseudomonas aeruginosa"
     variation       487..488
                     /replace="c"
                     /note="C reported in this position in original algD 
                     sequence"
     misc_feature    558..1868
                     /label=DNA from the TOL plasmid of Pseudomonas putida
                     /note="DNA from the TOL plasmid of Pseudomonas putida"
     variation       573
                     /replace="g"
                     /label=C in originally reported sequence
                     /note="C in originally reported sequence"
     variation       585
                     /replace="g"
                     /label=C in originally reported sequence
                     /note="C in originally reported sequence"
     gene            651..1588
                     /gene="xylE"
                     /label=xylE
     regulatory      651..657
                     /regulatory_class="ribosome_binding_site"
                     /gene="xylE"
     CDS             665..1588
                     /codon_start=1
                     /gene="xylE"
                     /product="catechol-2,3-dioxygenase"
                     /label=xylE
                     /protein_id="AAB60532.1"
                     /translation="MNKGVMRPGHVQLRVLDMSKALEHYVELLGLIEMDRDDQGRVYLK
                     AWTEVDKFSLVLREADEPGMDFMGFKVVDEDALRQLERDLMAYGCAVEQLPAGELNSCG
                     RRVRFQAPSGHHFELYADKEYTGKWGLNDVNPEAWPRDLKGMAAVRFDHALMYGDELPA
                     TYDLFTKVLGFYLAEQVLDENGTRVAQFLSLSTKAHDVAFIHHPEKGRLHHVSFHLETW
                     EDLLRAADLISMTDTSIDIGPTRHGLTHGKTIYFFDPSGNRNEVFCGGDYNYPDHKPVT
                     WTTDQLGKAIFYHDRILNERFMTVLT"
     misc_feature    1887..1943
                     /standard_name="polylinker"
                     /label=polylinker
     primer_bind     complement(1956..1972)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1980..1996
                     /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(2004..2034)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2049..2070
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2358..2946)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     gene            complement(3117..3987)
                     /gene="bla"
                     /label=bla
     CDS             complement(3117..3977)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="AAB60533.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3978..4082)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      3983..3987
                     /regulatory_class="ribosome_binding_site"