LOCUS       40924_7261        6628 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Yeast integrative vector pBS47, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6628)
  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  JOURNAL   Unpublished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 6628)
  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 6628)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6628)
  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            "Unpublished"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..6628
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        11..398
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             434..454
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             464..484
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             488..508
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             518..1234
                     /codon_start=1
                     /label=EYFP
                     /note="enhanced YFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(2094..2110)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      2159..2348
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(2371..2389)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2410..2426)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2434..2450)
                     /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(2458..2488)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2503..2524)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2812..3400)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3574..4431)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4432..4536)
                     /label=AmpR promoter
     promoter        4821..5041
                     /label=URA3 promoter
     CDS             5042..5842
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(5974..6429)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     6574..6590
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6597..6615
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        6628
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
ORIGIN
        1 ccagcttcaa aatgtttcta ctcctttttt actcttccag attttctcgg actccgcgca
       61 tcgccgtacc acttcaaaac acccaagcac agcatactaa atttcccctc tttcttcctc
      121 tagggtgtcg ttaattaccc gtactaaagg tttggaaaag aaaaaagaga ccgcctcgtt
      181 tctttttctt cgtcgaaaaa ggcaataaaa atttttatca cgtttctttt tcttgaaaat
      241 tttttttttg atttttttct ctttcgatga cctcccattg atatttaagt taataaacgg
      301 tcttcaattt ctcaagtttc agtttcattt ttcttgttct attacaactt tttttacttc
      361 ttgctcatta gaaagaaagc atagcaatct aatctaagtt ttctagatat ctgcagaatt
      421 cgcccttatg gttccaaaga agaagagaaa ggttttaatt gatccaaaaa agaaaagaaa
      481 ggttgatcca aagaaaaaga gaaaagtttt aattaacatg gtgagtaaag gagaagaact
      541 tttcactgga gttgtcccaa ttcttgttga attagatggt gatgttaatg ggcacaaatt
      601 ttctgtcagt ggagagggtg aaggtgatgc aacatacgga aaacttaccc ttaaatttat
      661 ttgcactact ggaaaactac ctgttccatg gccaacactt gtcactactt tcggttatgg
      721 tctaaaatgc tttgctagat acccagatca tatgaaacgg catgactttt tcaagagtgc
      781 catgcccgaa ggttatgtac aggaaagaac tatatttttc aaagatgacg ggaactacaa
      841 gacacgtgct gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg agttaaaagg
      901 tattgatttt aaagaagatg gaaacattct tggacacaaa ttggaataca actataactc
      961 acacaatgta tacatcatgg cagacaaaca aaagaatgga atcaaagtta acttcaaaat
     1021 tagacacaac attgaagatg gaagcgttca actagcagac cattatcaac aaaatactcc
     1081 aattggcgat ggccctgtcc ttttaccaga caaccattac ctgtcctatc aatctgccct
     1141 ttcgaaagat cccaacgaaa agagagacca catggtcctt cttgagtttg taacagctgc
     1201 tgggattaca catggcatgg atgaactata caaaggcgcc ccaccacctc ctccacctcc
     1261 accacctccc ggcgccagta aaggagaaga acttttcact ggagttgtcc caattcttgt
     1321 tgaattagat ggtgatgtta atgggcacaa attttctgtc agtggagagg gtgaaggtga
     1381 tgcaacatac ggaaaactta cccttaaatt tatttgcact actggaaaac tacctgttcc
     1441 atggccaaca cttgtcacta ctttgacttg gggtgttcaa tgcttttcaa gatacccaga
     1501 tcatatgaaa cggcatgact ttttcaagag tgccatgccc gaaggttatg tacaggaaag
     1561 aactatattt ttcaaagatg acgggaacta caagacacgt gctgaagtca agtttgaagg
     1621 tgataccctt gttaatagaa tcgagttaaa aggtattgat tttaaagaag atggaaacat
     1681 tcttggacac aaattggaat acaactatat ttcacacaat gtatacatca ctgcagacaa
     1741 acaaaagaat ggaatcaaag ctcatttcaa aattagacac aacattgaag atggaagcgt
     1801 tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg tccttttacc
     1861 agacaaccat tacctgtcca cacaatctgc cctttcgaaa gatcccaacg aaaagagaga
     1921 ccacatggtc cttcttgagt ttgtaacagc tgctgggatt acacatggca tggatgaact
     1981 atacaaatag ggcgcgccac ttctaaataa gcgaatttct tatgatttat aagggcgaat
     2041 tccagcacac tggcggccgt tactagtgga tccgagctcg gtaccaagct tatcgatacc
     2101 gtcgacctcg agtcatgtaa ttagttatgt cacgcttaca ttcacgccct ccccccacat
     2161 ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt
     2221 ttatagttat gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt
     2281 acagacgcgt gtacgcatgt aacattatac tgaaaacctt gcttgagaag gttttgggac
     2341 gctcgaaggc ttggtaccca gcttttgttc cctttagtga gggttaattc cgagcttggc
     2401 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     2461 cataggagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga ggtaactcac
     2521 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     2581 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     2641 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2701 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2761 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2821 gctcggcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2881 gacaggacta taaagatacc aggcgttccc ccctggaagc tccctcgtgc gctctcctgt
     2941 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3001 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3061 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3121 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3181 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3241 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3301 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3361 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3421 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3481 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3541 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3601 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctgcccgtcg tgtagataac
     3661 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3721 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3781 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3841 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3901 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3961 tacatgatcc cccatgttgt gaaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4021 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4081 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4141 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4201 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4261 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4321 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4381 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4441 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4501 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4561 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     4621 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     4681 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     4741 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     4801 actgagagtg caccacgctt ttcaattcaa ttcatcattt tttttttatt cttttttttg
     4861 atttcggttt ctttgaaatt tttttgattc ggtaatctcc gaacagaagg aagaacgaag
     4921 gaaggagcac agacttagat tggtatatat acgcatatgt agtgttgaag aaacatgaaa
     4981 ttgcccagta ttcttaaccc aactgcacag aacaaaaacc tgcaggaaac gaagataaat
     5041 catgtcgaaa gctacatata aggaacgtgc tgctactcat cctagtcctg ttgctgccaa
     5101 gctatttaat atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac
     5161 caccaaggaa ttactggagt tagttgaagc attaggtccc aaaatttgtt tactaaaaac
     5221 acatgtggat atcttgactg atttttccat ggagggcaca gttaagccgc taaaggcatt
     5281 atccgccaag tacaattttt tactcttcga agacagaaaa tttgctgaca ttggtaatac
     5341 agtcaaattg cagtactctg cgggtgtata cagaatagca gaatgggcag acattacgaa
     5401 tgcacacggt gtggtgggcc caggtattgt tagcggtttg aagcaggcgg cagaagaagt
     5461 aacaaaggaa cctagaggcc ttttgatgtt agcagaattg tcatgcaagg gctccctatc
     5521 tactggagaa tatactaagg gtactgttga cattgcgaag agcgacaaag attttgttat
     5581 cggctttatt gctcaaagag acatgggtgg aagagatgaa ggttacgatt ggttgattat
     5641 gacacccggt gtgggtttag atgacaaggg agacgcattg ggtcaacagt atagaaccgt
     5701 ggatgatgtg gtctctacag gatctgacat tattattgtt ggaagaggac tatttgcaaa
     5761 gggaagggat gctaaggtag agggtgaacg ttacagaaaa gcaggctggg aagcatattt
     5821 gagaagatgc ggccagcaaa actaaaaaac tgtattataa gtaaatgcat gtatactaaa
     5881 ctcacaaatt agagcttcaa tttaattata tcagttatta ccctgcggtg tgaaataccg
     5941 cacagatgcg taaggagaaa ataccgcatc aggaaattgt aaacgttaat attttgttaa
     6001 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     6061 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     6121 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     6181 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     6241 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     6301 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     6361 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     6421 agggcgcgtc gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     6481 gggcctcttc gctattacgc cagctggcga aggggggatg tgctgcaagg cgattaagtt
     6541 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattgtaat
     6601 acgactcact atagggcgaa ttggagct
//
