LOCUS       40924_20631        6229 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast integrative vector pFTR117-CFP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6229)
  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  JOURNAL   Unpublished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 6229)
  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 6229)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6229)
  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            "Unpublished"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..6229
                     /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             910..1620
                     /codon_start=1
                     /label=CFP
                     /note="cyan variant of GFP"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKAH
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(1727..1743)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      1792..1981
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(2004..2022)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2043..2059)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2067..2083)
                     /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(2091..2121)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2136..2157)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2445..3033)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3207..4064)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4065..4169)
                     /label=AmpR promoter
     promoter        4454..4674
                     /label=URA3 promoter
     CDS             4675..5475
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(5607..6062)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     6207..6223
                     /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 tgaccagcag tgacacccag aacaacaaaa cactggccgc aatgaaaaat
      601 tttgcggagc agtatgctaa acgtacggat acctattttt gtagtgatct ttccgtcacc
      661 gccgttgtta tcgagggttt agctcgccat aaagaggaat tgggttctcc cctgtgcccc
      721 tgtcgccact acgaagacaa ggaagcggaa gtaaaaaata ccttttggaa ttgcccctgt
      781 gtgcctatgc gggagcgcaa ggaatgtcac tgtatgttgt ttttaacccc agataacgat
      841 tttgctgggg atgcccagga tattcccatg gaaactttgg aggaagtcaa ggccagcatg
      901 ttaattaaca gtaaaggaga agaacttttc actggagttg tcccaattct tgttgaatta
      961 gatggtgatg ttaatgggca caaattttct gtcagtggag agggtgaagg tgatgcaaca
     1021 tacggaaaac ttacccttaa atttatttgc actactggaa aactacctgt tccatggcca
     1081 acacttgtca ctactttgac ttggggtgtt caatgctttt caagataccc agatcatatg
     1141 aaacggcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga aagaactata
     1201 tttttcaaag atgacgggaa ctacaagaca cgtgctgaag tcaagtttga aggtgatacc
     1261 cttgttaata gaatcgagtt aaaaggtatt gattttaaag aagatggaaa cattcttgga
     1321 cacaaattgg aatacaacta tatttcacac aatgtataca tcactgcaga caaacaaaag
     1381 aatggaatca aagctcattt caaaattaga cacaacattg aagatggaag cgttcaacta
     1441 gcagaccatt atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt accagacaac
     1501 cattacctgt ccacacaatc tgccctttcg aaagatccca acgaaaagag agaccacatg
     1561 gtccttcttg agtttgtaac agctgctggg attacacatg gcatggatga actatacaaa
     1621 tagggcgcgc cacttctaaa taagcgaatt tcttatgatt tataagggcg aattccagca
     1681 cactggcggc cgttactagt ggatccgagc tcggtaccaa gcttatcgat accgtcgacc
     1741 tcgagtcatg taattagtta tgtcacgctt acattcacgc cctcccccca catccgctct
     1801 aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt tttttatagt
     1861 tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc tgtacagacg
     1921 cgtgtacgca tgtaacatta tactgaaaac cttgcttgag aaggttttgg gacgctcgaa
     1981 ggcttggtac ccagcttttg ttccctttag tgagggttaa ttccgagctt ggcgtaatca
     2041 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatagga
     2101 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgaggtaact cacattaatt
     2161 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     2221 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     2281 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2341 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2401 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctcggc
     2461 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2521 ctataaagat accaggcgtt cccccctgga agctccctcg tgcgctctcc tgttccgacc
     2581 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa
     2641 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2701 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2761 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2821 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2881 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2941 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3001 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3061 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3121 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3181 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3241 atctgtctat ttcgttcatc catagttgcc tgactgcccg tcgtgtagat aactacgata
     3301 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     3361 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3421 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3481 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3541 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3601 tcccccatgt tgtgaaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3661 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3721 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3781 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3841 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3901 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3961 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4021 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4081 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     4141 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     4201 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     4261 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     4321 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     4381 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     4441 gtgcaccacg cttttcaatt caattcatca tttttttttt attctttttt ttgatttcgg
     4501 tttctttgaa atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag
     4561 cacagactta gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca
     4621 gtattcttaa cccaactgca cagaacaaaa acctgcagga aacgaagata aatcatgtcg
     4681 aaagctacat ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt
     4741 aatatcatgc acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag
     4801 gaattactgg agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg
     4861 gatatcttga ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc
     4921 aagtacaatt ttttactctt cgaagacaga aaatttgctg acattggtaa tacagtcaaa
     4981 ttgcagtact ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac
     5041 ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag
     5101 gaacctagag gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga
     5161 gaatatacta agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt
     5221 attgctcaaa gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc
     5281 ggtgtgggtt tagatgacaa gggagacgca ttgggtcaac agtatagaac cgtggatgat
     5341 gtggtctcta caggatctga cattattatt gttggaagag gactatttgc aaagggaagg
     5401 gatgctaagg tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga
     5461 tgcggccagc aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa
     5521 attagagctt caatttaatt atatcagtta ttaccctgcg gtgtgaaata ccgcacagat
     5581 gcgtaaggag aaaataccgc atcaggaaat tgtaaacgtt aatattttgt taaaattcgc
     5641 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     5701 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     5761 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     5821 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     5881 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     5941 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     6001 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     6061 gtcgcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     6121 ttcgctatta cgccagctgg cgaagggggg atgtgctgca aggcgattaa gttgggtaac
     6181 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattg
//
