LOCUS       Exported                8888 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCIB2, complete sequence.
ACCESSION   JX262692
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8888)
  AUTHORS   Dequard-Chablat M, Nguyen T-T., Contamine V, Hermann-Le Denmat S, 
            Malagnac F.
  TITLE     Efficient tools to target DNA to Podospora anserina
  JOURNAL   Fungal Genet Rep 59, 21-25 (2012)
REFERENCE   2  (bases 1 to 8888)
  AUTHORS   Dequard-Chablat M, Nguyen T-T., Contamine V, Hermann-Le Denmat S, 
            Malagnac F.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-JUN-2012) IGM, UPS XI, CNRS-UMR 8621, 15 rue Georges 
            Clemenceau, Orsay, Ile de France 91405, France
REFERENCE   3  (bases 1 to 8888)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8888
                     /organism="Cloning vector pCIB2"
                     /lab_host="Escherichia coli; Saccharomyces cerevisiae"
                     /mol_type="other DNA"
                     /note="used for targeting integration in Podospora 
                     anserina"
                     /db_xref="taxon:1211107"
     misc_feature    1..1998
                     /label=cloning vector pRS426
                     /note="cloning vector pRS426"
     promoter        201..416
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             417..1220
                     /codon_start=1
                     /gene="URA3"
                     /product="orotidine-5phosphate decarboxylase"
                     /label=URA3
                     /protein_id="AFP20537.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     gene            417..1220
                     /gene="URA3"
                     /label=URA3
                     /note="ScURA3"
     rep_origin      complement(1351..1806)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     1951..1967
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1977..1995
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1995..2002
                     /label=modified AscI restriction site
                     /note="modified AscI restriction site"
     5UTR           2003..2919
                     /gene="Pa_2_3690"
     gene            2003..2919
                     /gene="Pa_2_3690"
                     /label=Pa_2_3690
                     /note="from Podospora anserina chromosome 2"
     misc_feature    2920..2927
                     /label=modified SwaI restriction site
                     /note="modified SwaI restriction site"
     regulatory      2928..3263
                     /regulatory_class="promoter"
                     /gene="GPD"
                     /gene_synonym="Pa_3_5110"
     gene            2928..3263
                     /gene="GPD"
                     /gene_synonym="Pa_3_5110"
                     /label=GPD
     CDS             3264..3836
                     /codon_start=1
                     /gene="Nat1"
                     /product="nourseothricin acetyl transferase"
                     /function="nourseothricin resistance"
                     /label=Nat1
                     /protein_id="AFP20536.1"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     gene            3264..3836
                     /gene="Nat1"
                     /label=Nat1
     regulatory      3886..4275
                     /regulatory_class="terminator"
                     /gene="AS1"
                     /gene_synonym="Pa_1_16650"
     gene            3886..4275
                     /gene="AS1"
                     /gene_synonym="Pa_1_16650"
                     /label=AS1
     misc_feature    4276..4283
                     /label=modified PacI restriction site
                     /note="modified PacI restriction site"
     3UTR           4283..5250
                     /gene="Pa_2_3690"
     gene            4283..5250
                     /gene="Pa_2_3690"
                     /label=Pa_2_3690
                     /note="from Podospora anserina chromosome 2"
     misc_feature    5251..5258
                     /label=modified AscI restriction site
                     /note="modified AscI restriction site"
     misc_feature    5255..8888
                     /label=cloning vector pRS426
                     /note="cloning vector pRS426"
     promoter        complement(5286..5304)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5325..5341)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5349..5365
                     /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(5373..5403)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(5727..6315)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6486..7346)
                     /codon_start=1
                     /gene="BLA"
                     /product="Beta-lactamase"
                     /label=BLA
                     /protein_id="AFP20538.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(6486..7346)
                     /gene="BLA"
                     /label=BLA
                     /note="ampicillin resistance"
     promoter        complement(7347..7451)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7478..8820
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(7478..8449)
                     /direction=LEFT
                     /label=2-micron origin
                     /note="2-micron origin"
     protein_bind    complement(8441..8488)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."