LOCUS       40924_20636        7234 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast integrative vector pFTR74-YFP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7234)
  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  JOURNAL   Unpublished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 7234)
  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 7234)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7234)
  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            "Unpublished"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..7234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        34..430
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             466..486
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             496..516
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             520..540
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             784..1494
                     /codon_start=1
                     /label=EYFP
                     /note="enhanced YFP"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(1601..1617)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      1666..1855
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(1878..1896)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1917..1933)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1941..1957)
                     /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(1965..1995)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2010..2031)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2319..2907)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3081..3938)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3939..4043)
                     /label=AmpR promoter
     CDS             complement(4804..5895)
                     /codon_start=1
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(5908..6312)
                     /label=LEU2 promoter
     rep_origin      complement(6612..7067)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     7212..7228
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg cgaattggag ctccagcttc aaaatgtttc tactcctttt
       61 ttactcttcc agattttctc ggactccgcg catcgccgta ccacttcaaa acacccaagc
      121 acagcatact aaatttcccc tctttcttcc tctagggtgt cgttaattac ccgtactaaa
      181 ggtttggaaa agaaaaaaga gaccgcctcg tttctttttc ttcgtcgaaa aaggcaataa
      241 aaatttttat cacgtttctt tttcttgaaa attttttttt tgattttttt ctctttcgat
      301 gacctcccat tgatatttaa gttaataaac ggtcttcaat ttctcaagtt tcagtttcat
      361 ttttcttgtt ctattacaac tttttttact tcttgctcat tagaaagaaa gcatagcaat
      421 ctaatctaag ttttctagat atctgcagaa ttcgccctta tggttccaaa gaagaagaga
      481 aaggttttaa ttgatccaaa aaagaaaaga aaggttgatc caaagaaaaa gagaaaagtt
      541 ttaattaaca tgaatgtcgg cgatcgtgtt cgagtcacct cctccgttgt ggtttaccac
      601 catccagaac ataaaaaaac agcctttgat ttacaaggta tggaagggga agtggccgcc
      661 gtattgaccg aatggcaggg caggcccatc agcgctaatt tgccggtttt agtcaagttt
      721 gaacagcgtt ttaaggccca tttccgtccg gatgaagtta ccctgattga agatttaatt
      781 aacagtaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt
      841 gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc aacatacgga
      901 aaacttaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg gccaacactt
      961 gtcactactt tcggttatgg tctaaaatgc tttgctagat acccagatca tatgaaacgg
     1021 catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaaagaac tatatttttc
     1081 aaagatgacg ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga tacccttgtt
     1141 aatagaatcg agttaaaagg tattgatttt aaagaagatg gaaacattct tggacacaaa
     1201 ttggaataca actataactc acacaatgta tacatcatgg cagacaaaca aaagaatgga
     1261 atcaaagtta acttcaaaat tagacacaac attgaagatg gaagcgttca actagcagac
     1321 cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac
     1381 ctgtcctatc aatctgccct ttcgaaagat cccaacgaaa agagagacca catggtcctt
     1441 cttgagtttg taacagctgc tgggattaca catggcatgg atgaactata caaatagggc
     1501 gcgccacttc taaataagcg aatttcttat gatttataag ggcgaattcc agcacactgg
     1561 cggccgttac tagtggatcc gagctcggta ccaagcttat cgataccgtc gacctcgagt
     1621 catgtaatta gttatgtcac gcttacattc acgccctccc cccacatccg ctctaaccga
     1681 aaaggaagga gttagacaac ctgaagtcta ggtccctatt tattttttta tagttatgtt
     1741 agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca gacgcgtgta
     1801 cgcatgtaac attatactga aaaccttgct tgagaaggtt ttgggacgct cgaaggcttg
     1861 gtacccagct tttgttccct ttagtgaggg ttaattccga gcttggcgta atcatggtca
     1921 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat aggagccgga
     1981 agcataaagt gtaaagcctg gggtgcctaa tgagtgaggt aactcacatt aattgcgttg
     2041 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     2101 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     2161 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2221 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2281 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct cggcccccct
     2341 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2401 agataccagg cgttcccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2461 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     2521 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2581 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2641 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     2701 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     2761 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     2821 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     2881 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     2941 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3001 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3061 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3121 ctatttcgtt catccatagt tgcctgactg cccgtcgtgt agataactac gatacgggag
     3181 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3241 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3301 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3361 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     3421 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     3481 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     3541 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     3601 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     3661 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     3721 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     3781 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     3841 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     3901 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     3961 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4021 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     4081 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     4141 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     4201 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     4261 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     4321 catatcgact acgtcgtaag gccgtttctg acagagtaaa attcttgagg gaactttcac
     4381 cattatggga aatggttcaa gaaggtattg acttaaactc catcaaatgg tcaggtcatt
     4441 gagtgttttt tatttgttgt attttttttt ttttagagaa aatcctccaa tatcaaatta
     4501 ggaatcgtag tttcatgatt ttctgttaca cctaactttt tgtgtggtgc cctcctcctt
     4561 gtcaatatta atgttaaagt gcaattcttt ttccttatca cgttgagcca ttagtatcaa
     4621 tttgcttacc tgtattcctt tactatcctc ctttttctcc ttcttgataa atgtatgtag
     4681 attgcgtata tagtttcgtc taccctatga acatattcca ttttgtaatt tcgtgtcgtt
     4741 tctattatga atttcattta taaagtttat gtacaaatat cataaaaaaa gagaatcttt
     4801 ttaagcaagg attttcttaa cttcttcggc gacagcatca ccgacttcgg tggtactgtt
     4861 ggaaccacct aaatcaccag ttctgatacc tgcatccaaa acctttttaa ctgcatcttc
     4921 aatggcctta ccttcttcag gcaagttcaa tgacaatttc aacatcattg cagcagacaa
     4981 gatagtggcg atagggtcaa ccttattctt tggcaaatct ggagcagaac cgtggcatgg
     5041 ttcgtacaaa ccaaatgcgg tgttcttgtc tggcaaagag gccaaggacg cagatggcaa
     5101 caaacccaag gaacctggga taacggaggc ttcatcggag atgatatcac caaacatgtt
     5161 gctggtgatt ataataccat ttaggtgggt tgggttctta actaggatca tggcggcaga
     5221 atcaatcaat tgatgttgaa ccttcaatgt agggaattcg ttcttgatgg tttcctccac
     5281 agtttttctc cataatcttg aagaggccaa aacattagct ttatccaagg accaaatagg
     5341 caatggtggc tcatgttgta gggccatgaa agcggccatt cttgtgattc tttgcacttc
     5401 tggaacggtg tattgttcac tatcccaagc gacaccatca ccatcgtctt cctttctctt
     5461 accaaagtaa atacctccca ctaattctct gacaacaacg aagtcagtac ctttagcaaa
     5521 ttgtggcttg attggagata agtctaaaag agagtcggat gcaaagttac atggtcttaa
     5581 gttggcgtac aattgaagtt ctttacggat ttttagtaaa ccttgttcag gtctaacact
     5641 accggtaccc catttaggac cacccacagc acctaacaaa acggcatcaa ccttcttgga
     5701 ggcttccagc gcctcatctg gaagtgggac acctgtagca tcgatagcag caccaccaat
     5761 taaatgattt tcgaaatcga acttgacatt ggaacgaaca tcagaaatag ctttaagaac
     5821 cttaatggct tcggctgtga tttcttgacc aacgtggtca cctggcaaaa cgacgatctt
     5881 cttaggggca gacatagggg cagacattag aatggtatat ccttgaaata tatatatata
     5941 ttgctgaaat gtaaaaggta agaaaagtta gaaagtaaga cgattgctaa ccacctattg
     6001 gaaaaaacaa taggtcctta aataatattg tcaacttcaa gtattgtgat gcaagcattt
     6061 agtcatgaac gcttctctat tctatatgaa aagccggttc cggcctctca cctttccttt
     6121 ttctcccaat ttttcagttg aaaaaggtat atgcgtcagg cgacctctga aattaacaaa
     6181 aaatttccag tcatcgaatt tgattctgtg cgatagcgcc cctgtgtgtt ctcgttatgt
     6241 tgaggaaaaa aataatggtt gctaagagat tcgaactctt gcatcttacg atacctgagt
     6301 attcccacag ttaactgcgg tcaagatatt tcttgaatca ggcgccttag accgctcggc
     6361 caaacaacca attacttgtt gagaaataga gtataattat cctataaata taacgttttt
     6421 gaacacacat gaacaaggaa gtacaggaca attgattttg aagagaatgt ggattttgat
     6481 gtaattgttg ggattccatt tttaataagg caataatatt aggtatgtgg atatactaga
     6541 agttctcctc gaccgtcgat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     6601 accgcatcag gaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     6661 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     6721 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     6781 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     6841 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     6901 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     6961 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     7021 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcgc gccattcgcc
     7081 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     7141 gctggcgaag gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     7201 gtcacgacgt tgtaaaacga cggccagtga attg
//
