LOCUS       Exported                6628 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pW1336, complete sequence.
ACCESSION   KT033769
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6628)
  AUTHORS   Barrett SP, Wang PL, Salzman J.
  TITLE     Circular RNA biogenesis can proceed through an exon-containing 
            lariat precursor
  JOURNAL   Elife 4 (2015) In press
  PUBMED    26057830
REFERENCE   2  (bases 1 to 6628)
  AUTHORS   Wang PL.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-JUN-2015) Biochemistry, Stanford University, 279 
            Campus Drive, Stanford, CA 94305, USA
REFERENCE   3  (bases 1 to 6628)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6628
                     /organism="Cloning vector pW1336"
                     /mol_type="other DNA"
                     /db_xref="taxon:1670968"
     rep_origin      74..1279
                     /label=ars1
                     /note="ars1"
     rep_origin      147..930
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     primer_bind     complement(1280..1296)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      1491..1966
                     /label=M13 origin
                     /note="M13 origin"
     rep_origin      1509..1964
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2246..2350
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2351..3211
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="AKM50016.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3382..3970
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4258..4279
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4294..4324
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4332..4348
                     /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)."
     primer_bind     4356..4372
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     gene            complement(4385..6622)
                     /gene="LEU2"
                     /label=LEU2
     CDS             complement(4873..5967)
                     /codon_start=1
                     /gene="LEU2"
                     /product="beta-isopropylmalate dehydrogenase"
                     /label=LEU2
                     /note="derived from Saccharomyces cerevisiae"
                     /protein_id="AKM50017.1"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(5968..6375)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     misc_feature    6623..73
                     /note="multiple cloning site: SphI, SpeI, SfiI, NcoI, AscI,
                     PacI, NotI, XhoI, SacI"