LOCUS       Exported                8786 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCHC01, complete sequence.
ACCESSION   HM014235
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8786)
  AUTHORS   Chen C-H., DeMay BS, Gladfelter AS, Dunlap JC, Loros JJ.
  TITLE     Direct interaction between VVD and WCC regulates photoadaptation in 
            Neurospora
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8786)
  AUTHORS   Chen C-H.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-MAR-2010) Genetics, Dartmouth Medical School, 702 
            Remsen, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 8786)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8786
                     /organism="Cloning vector pCHC01"
                     /mol_type="other DNA"
                     /db_xref="taxon:864788"
     promoter        201..416
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             417..1220
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(1351..1806)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     misc_recomb     1957..3025
                     /note="derived from Neurospora crassa csr-1; Neurospora 
                     homologous recombination site"
     misc_recomb     3026..3054
                     /label=yeast homologous recombination site
                     /note="yeast homologous recombination site"
     promoter        3055..3072
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_recomb     3085..3113
                     /label=yeast homologous recombination site
                     /note="yeast homologous recombination site"
     CDS             complement(3261..3812)
                     /codon_start=1
                     /product="Bar"
                     /label=Bar
                     /note="herbicide resistance protein"
                     /protein_id="ADK36683.1"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     misc_recomb     4206..5220
                     /note="derived from Neurospora crassa csr-1; Neurospora 
                     homologous recombination site"
     protein_bind    5247..5263
                     /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(5271..5301)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(5625..6213)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6384..7244)
                     /codon_start=1
                     /product="AmpR"
                     /label=AmpR
                     /protein_id="ADK36682.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7245..7349)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7376..8718
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(8339..8386)
                     /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)."