LOCUS       Exported                6238 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pDblet, complete sequence.
ACCESSION   AF531173
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6238)
  AUTHORS   Zhu J, Carlson DL, Dubey DD, Sharma K, Huberman JA.
  TITLE     Comparison of the two major ARS elements of the ura4 replication 
            origin region with other ARS elements in the fission yeast, 
            Schizosaccharomyces pombe
  JOURNAL   Chromosoma 103 (6), 414-422 (1994)
  PUBMED    7859562
REFERENCE   2  (bases 1 to 6238)
  AUTHORS   Brun C, Dubey DD, Huberman JA.
  TITLE     pDblet, a stable autonomously replicating shuttle vector for 
            Schizosaccharomyces pombe
  JOURNAL   Gene 164 (1), 173-177 (1995)
  PUBMED    7590311
REFERENCE   3  (bases 1 to 6238)
  AUTHORS   Brun C, Dubey DD, Huberman JA.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-JUL-2002) Cancer Genetics, Roswell Park Cancer 
            Institute, Elm & Carlton Streets, Buffalo, NY 14263-0001, USA
REFERENCE   4  (bases 1 to 6238)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6238
                     /organism="Cloning vector pDblet"
                     /focus
                     /mol_type="genomic DNA"
                     /note="replicates efficiently in Schizosaccharomyces pombe;
                     vector is mostly pUC19"
                     /db_xref="taxon:204048"
     source          186..1955
                     /organism="Schizosaccharomyces pombe"
                     /mol_type="genomic DNA"
                     /label=HindIII fragment
                     /note="HindIII fragment"
                     /db_xref="taxon:4896"
     source          2007..2462
                     /organism="Enterobacteria phage f1"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:10863"
     source          2537..2984
                     /organism="Cloning vector pBluescript"
                     /mol_type="genomic DNA"
                     /label=pBluescript II SK+ (SYNBL2SKP)
                     /note="pBluescript II SK+ (SYNBL2SKP)"
                     /db_xref="taxon:42057"
     source          4974..6171
                     /organism="Schizosaccharomyces pombe"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:4896"
     CDS             complement(625..1419)
                     /codon_start=1
                     /gene="ura4"
                     /product="OMP decarboxylase"
                     /label=ura4
                     /note="component of uracil biosynthesis pathway"
                     /protein_id="AAM95949.1"
                     /translation="MDARVFQSYSARAEGMKNPIAKELLALMEEKQSNLSVAVDLTKKS
                     EILELVDKIGPYVCVIKTHIDVVEDFDQDMVEKLVALGKKHRFLIFEDRKFADIGNTVK
                     LQYASGVYKIASWAHITNCHTVPGEGIIQGLKEVGLPLGRGLLLLAEMSSKGSLATGSY
                     TEKTLEWFEKHTDFCFGFIAGRRFPNLQSDYITMSPGIGLDVKGDGLGQQYRTPEEVIV
                     NCGSDIIIVGRGVYGAGRNPVVEAKRYREAGWKAYQQRLSQH"
     gene            complement(625..1419)
                     /gene="ura4"
                     /label=ura4
     rep_origin      complement(2007..2462)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(2245..2823)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFA
                     GFPRQALNRGLPLGFRFSALRHLDPKKLD"
     primer_bind     2607..2623
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2633..2651
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2660..2767
                     /label=multiple cloning site
                     /note="multiple cloning site"
     primer_bind     2677..2693
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2727..2743)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2780..2798)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2819..2835)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2843..2859
                     /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(2867..2897)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2912..2933
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3221..3809)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3980..4840)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="AAM95950.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4841..4945)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4974..6171
                     /label=ARS doublet bordered by AatII sites
                     /note="ARS doublet bordered by AatII sites"