LOCUS       Exported                6752 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast integrative vector pYFP-Lim4-CFP, complete sequence.
ACCESSION   KF177464
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6752)
  AUTHORS   Bonomi M, Muller EGD., Pellarin R, Kim SJ, Russel D, Ramsden R, 
            Sundin BA, Davis TN, Sali A.
  TITLE     Protein complex structures from Bayesian modeling of in vivo FRET 
            data
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6752)
  AUTHORS   Bonomi M, Muller EGD., Pellarin R, Kim SJ, Russel D, Ramsden R, 
            Sundin BA, Davis TN, Sali A.
  TITLE     Direct Submission
  JOURNAL   Submitted (31-MAY-2013) Biochemistry, University of Washington, 1705
            NE Pacific St, Seattle, WA 98195-7350, USA
REFERENCE   3  (bases 1 to 6752)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: CLC Main Workbench v. 6
            Sequencing Technology :: Sanger dideoxy sequencing; Genbank 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6752
                     /organism="Yeast integrative vector pYFP-Lim4-CFP"
                     /mol_type="other DNA"
                     /db_xref="taxon:1400784"
     terminator      850..1039
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(1062..1080)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1101..1117)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1125..1141
                     /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(1149..1179)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      1447..2114
                     /label=pUC
                     /note="pUC"
     rep_origin      complement(1503..2091)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2262..3122)
                     /codon_start=1
                     /product="AmpR"
                     /label=AmpR
                     /note="ampicillin resistance protein"
                     /protein_id="AGW21638.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3123..3227)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3517..3732
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             3733..4536
                     /codon_start=1
                     /product="URA3"
                     /label=URA3
                     /protein_id="AGW21639.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(4665..5120)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     5265..5281
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5288..5306
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      5321..5720
                     /regulatory_class="promoter"
                     /note="TEF"
     CDS             5747..722
                     /codon_start=1
                     /product="NLS-YFP-Lim4-CFP"
                     /label=NLS-YFP-Lim4-CFP
                     /note="Lim4 linked to YFP and CFP with a nuclear 
                     localization signal; adaptor protein at focal adhesions"
                     /protein_id="AGW21640.1"
                     /translation="MVPKKKRKVLIDPKKKRKVDPKKKRKVLINMVSKGEELFTGVVPI
                     LVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFGYGLKCFAR
                     YPDHMKRHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKE
                     DGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDG
                     PVLLPDNHYLSYQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYKGGSGVPICGACR
                     RPIEGRVVNAMGKQWHVEHFVCAKCEKPFLGHRHYERKGLAYCETHYNQLFGDVASASK
                     GEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVT
                     TLTWGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVN
                     RIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKAHFKIRHNIEDGSVQLAD
                     HYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYK"
     CDS             5753..5773
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5783..5803
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5807..5827
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6596..6607
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"