LOCUS       40924_48093        6833 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast integrative vector pYFP-TAF28-CFP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6833)
  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  JOURNAL   Unpublished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 6833)
  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 6833)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6833)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (31-MAY-2013) Biochemistry, University of Washington, 1705 NE 
            Pacific St, Seattle, WA 98195-7350, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
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..6833
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..20
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        35..431
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             467..487
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             497..517
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             521..541
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             551..1267
                     /codon_start=1
                     /label=EYFP
                     /note="enhanced YFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     terminator      2397..2586
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(2609..2627)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2648..2664)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2672..2688)
                     /label=lac operator
                     /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(2696..2726)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2741..2762)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3050..3638)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3812..4669)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4670..4774)
                     /label=AmpR promoter
     promoter        5059..5279
                     /label=URA3 promoter
     CDS             5280..6080
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(6212..6667)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     6812..6828
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gtaatacgac tcactatagg gcgaattgga gctccagctt caaaatgttt ctactccttt
       61 tttactcttc cagattttct cggactccgc gcatcgccgt accacttcaa aacacccaag
      121 cacagcatac taaatttccc ctctttcttc ctctagggtg tcgttaatta cccgtactaa
      181 aggtttggaa aagaaaaaag agaccgcctc gtttcttttt cttcgtcgaa aaaggcaata
      241 aaaattttta tcacgtttct ttttcttgaa aatttttttt ttgatttttt tctctttcga
      301 tgacctccca ttgatattta agttaataaa cggtcttcaa tttctcaagt ttcagtttca
      361 tttttcttgt tctattacaa ctttttttac ttcttgctca ttagaaagaa agcatagcaa
      421 tctaatctaa gttttctaga tatctgcaga attcgccctt atggttccaa agaagaagag
      481 aaaggtttta attgatccaa aaaagaaaag aaaggttgat ccaaagaaaa agagaaaagt
      541 tttaattaac atggtgagta aaggagaaga acttttcact ggagttgtcc caattcttgt
      601 tgaattagat ggtgatgtta atgggcacaa attttctgtc agtggagagg gtgaaggtga
      661 tgcaacatac ggaaaactta cccttaaatt tatttgcact actggaaaac tacctgttcc
      721 atggccaaca cttgtcacta ctttcggtta tggtctaaaa tgctttgcta gatacccaga
      781 tcatatgaaa cggcatgact ttttcaagag tgccatgccc gaaggttatg tacaggaaag
      841 aactatattt ttcaaagatg acgggaacta caagacacgt gctgaagtca agtttgaagg
      901 tgataccctt gttaatagaa tcgagttaaa aggtattgat tttaaagaag atggaaacat
      961 tcttggacac aaattggaat acaactataa ctcacacaat gtatacatca tggcagacaa
     1021 acaaaagaat ggaatcaaag ttaacttcaa aattagacac aacattgaag atggaagcgt
     1081 tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg tccttttacc
     1141 agacaaccat tacctgtcct atcaatctgc cctttcgaaa gatcccaacg aaaagagaga
     1201 ccacatggtc cttcttgagt ttgtaacagc tgctgggatt acacatggca tggatgaact
     1261 atacaaaggc ggatccatgt tttctgagga gcagctgaac cgttatgaaa tgtatcgccg
     1321 ctcagccttc cctaaggcag ccatcaaaag gctgatccag tccatcactg gcacctctgt
     1381 gtctcagaat gttgttattg ctatgtctgg tatttccaag gttttcgtcg gggaggtggt
     1441 agaagaagca ctggatgtgt gtgagaagtg gggagaaatg ccaccactac aacccaaaca
     1501 tatgagggaa gccgttagaa ggttaaagtc aaaaggacag atccctgcta gcgccagtaa
     1561 aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa
     1621 tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac
     1681 ccttaaattt atttgcacta ctggaaaact acctgttcca tggccaacac ttgtcactac
     1741 tttgacttgg ggtgttcaat gcttttcaag atacccagat catatgaaac ggcatgactt
     1801 tttcaagagt gccatgcccg aaggttatgt acaggaaaga actatatttt tcaaagatga
     1861 cgggaactac aagacacgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat
     1921 cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt cttggacaca aattggaata
     1981 caactatatt tcacacaatg tatacatcac tgcagacaaa caaaagaatg gaatcaaagc
     2041 tcatttcaaa attagacaca acattgaaga tggaagcgtt caactagcag accattatca
     2101 acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccac
     2161 acaatctgcc ctttcgaaag atcccaacga aaagagagac cacatggtcc ttcttgagtt
     2221 tgtaacagct gctgggatta cacatggcat ggatgaacta tacaaatagg gcgcgccact
     2281 tctaaataag cgaatttctt atgatttata agggcgaatt ccagcacact ggcggccgtt
     2341 actagtcgag tcatgtaatt agttatgtca cgcttacatt cacgccctcc ccccacatcc
     2401 gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat ttattttttt
     2461 atagttatgt tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac
     2521 agacgcgtgt acgcatgtaa cattatactg aaaaccttgc ttgagaaggt tttgggacgc
     2581 tcgaaggctt ggtacccagc ttttgttccc tttagtgagg gttaattccg agcttggcgt
     2641 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     2701 taggagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagg taactcacat
     2761 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     2821 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     2881 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     2941 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3001 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3061 tcggcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3121 caggactata aagataccag gcgttccccc ctggaagctc cctcgtgcgc tctcctgttc
     3181 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3241 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3301 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3361 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3421 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3481 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3541 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3601 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3661 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     3721 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3781 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3841 cagcgatctg tctatttcgt tcatccatag ttgcctgact gcccgtcgtg tagataacta
     3901 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     3961 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     4021 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     4081 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     4141 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     4201 catgatcccc catgttgtga aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     4261 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     4321 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     4381 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     4441 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     4501 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4561 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     4621 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     4681 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     4741 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     4801 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     4861 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     4921 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     4981 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     5041 tgagagtgca ccacgctttt caattcaatt catcattttt tttttattct tttttttgat
     5101 ttcggtttct ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga
     5161 aggagcacag acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt
     5221 gcccagtatt cttaacccaa ctgcacagaa caaaaacctg caggaaacga agataaatca
     5281 tgtcgaaagc tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc
     5341 tatttaatat catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca
     5401 ccaaggaatt actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac
     5461 atgtggatat cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat
     5521 ccgccaagta caatttttta ctcttcgaag acagaaaatt tgctgacatt ggtaatacag
     5581 tcaaattgca gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg
     5641 cacacggtgt ggtgggccca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa
     5701 caaaggaacc tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta
     5761 ctggagaata tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg
     5821 gctttattgc tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga
     5881 cacccggtgt gggtttagat gacaagggag acgcattggg tcaacagtat agaaccgtgg
     5941 atgatgtggt ctctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg
     6001 gaagggatgc taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga
     6061 gaagatgcgg ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact
     6121 cacaaattag agcttcaatt taattatatc agttattacc ctgcggtgtg aaataccgca
     6181 cagatgcgta aggagaaaat accgcatcag gaaattgtaa acgttaatat tttgttaaaa
     6241 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     6301 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     6361 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     6421 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     6481 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     6541 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     6601 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     6661 ggcgcgtcgc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     6721 gcctcttcgc tattacgcca gctggcgaag gggggatgtg ctgcaaggcg attaagttgg
     6781 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga att
//
