LOCUS       40924_7276        6658 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Yeast integrative vector pBS50, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6658)
  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  JOURNAL   Unpublished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 6658)
  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 6658)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6658)
  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            "Unpublished"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..6658
                     /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(2124..2140)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      2189..2378
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(2401..2419)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2440..2456)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2464..2480)
                     /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(2488..2518)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2533..2554)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2842..3430)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3604..4461)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4462..4566)
                     /label=AmpR promoter
     promoter        4851..5071
                     /label=URA3 promoter
     CDS             5072..5872
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(6004..6459)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     6604..6620
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6627..6645
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        6658
                     /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 caaaggcgct ccaccacctc cacctccacc
     1261 accacctcca cctccaccac ctccacctcc acctccaccc ggcgccagta aaggagaaga
     1321 acttttcact ggagttgtcc caattcttgt tgaattagat ggtgatgtta atgggcacaa
     1381 attttctgtc agtggagagg gtgaaggtga tgcaacatac ggaaaactta cccttaaatt
     1441 tatttgcact actggaaaac tacctgttcc atggccaaca cttgtcacta ctttgacttg
     1501 gggtgttcaa tgcttttcaa gatacccaga tcatatgaaa cggcatgact ttttcaagag
     1561 tgccatgccc gaaggttatg tacaggaaag aactatattt ttcaaagatg acgggaacta
     1621 caagacacgt gctgaagtca agtttgaagg tgataccctt gttaatagaa tcgagttaaa
     1681 aggtattgat tttaaagaag atggaaacat tcttggacac aaattggaat acaactatat
     1741 ttcacacaat gtatacatca ctgcagacaa acaaaagaat ggaatcaaag ctcatttcaa
     1801 aattagacac aacattgaag atggaagcgt tcaactagca gaccattatc aacaaaatac
     1861 tccaattggc gatggccctg tccttttacc agacaaccat tacctgtcca cacaatctgc
     1921 cctttcgaaa gatcccaacg aaaagagaga ccacatggtc cttcttgagt ttgtaacagc
     1981 tgctgggatt acacatggca tggatgaact atacaaatag ggcgcgccac ttctaaataa
     2041 gcgaatttct tatgatttat aagggcgaat tccagcacac tggcggccgt tactagtgga
     2101 tccgagctcg gtaccaagct tatcgatacc gtcgacctcg agtcatgtaa ttagttatgt
     2161 cacgcttaca ttcacgccct ccccccacat ccgctctaac cgaaaaggaa ggagttagac
     2221 aacctgaagt ctaggtccct atttattttt ttatagttat gttagtatta agaacgttat
     2281 ttatatttca aatttttctt ttttttctgt acagacgcgt gtacgcatgt aacattatac
     2341 tgaaaacctt gcttgagaag gttttgggac gctcgaaggc ttggtaccca gcttttgttc
     2401 cctttagtga gggttaattc cgagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     2461 aaattgttat ccgctcacaa ttccacacaa cataggagcc ggaagcataa agtgtaaagc
     2521 ctggggtgcc taatgagtga ggtaactcac attaattgcg ttgcgctcac tgcccgcttt
     2581 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     2641 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2701 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2761 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2821 aaaggccgcg ttgctggcgt ttttccatag gctcggcccc cctgacgagc atcacaaaaa
     2881 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgttccc
     2941 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3001 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     3061 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3121 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3181 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3241 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     3301 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3361 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3421 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3481 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3541 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3601 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3661 agttgcctga ctgcccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3721 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3781 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3841 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3901 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3961 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gaaaaaaagc
     4021 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4081 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4141 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4201 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4261 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4321 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4381 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4441 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4501 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4561 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     4621 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     4681 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     4741 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     4801 gcttaactat gcggcatcag agcagattgt actgagagtg caccacgctt ttcaattcaa
     4861 ttcatcattt tttttttatt cttttttttg atttcggttt ctttgaaatt tttttgattc
     4921 ggtaatctcc gaacagaagg aagaacgaag gaaggagcac agacttagat tggtatatat
     4981 acgcatatgt agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc aactgcacag
     5041 aacaaaaacc tgcaggaaac gaagataaat catgtcgaaa gctacatata aggaacgtgc
     5101 tgctactcat cctagtcctg ttgctgccaa gctatttaat atcatgcacg aaaagcaaac
     5161 aaacttgtgt gcttcattgg atgttcgtac caccaaggaa ttactggagt tagttgaagc
     5221 attaggtccc aaaatttgtt tactaaaaac acatgtggat atcttgactg atttttccat
     5281 ggagggcaca gttaagccgc taaaggcatt atccgccaag tacaattttt tactcttcga
     5341 agacagaaaa tttgctgaca ttggtaatac agtcaaattg cagtactctg cgggtgtata
     5401 cagaatagca gaatgggcag acattacgaa tgcacacggt gtggtgggcc caggtattgt
     5461 tagcggtttg aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt
     5521 agcagaattg tcatgcaagg gctccctatc tactggagaa tatactaagg gtactgttga
     5581 cattgcgaag agcgacaaag attttgttat cggctttatt gctcaaagag acatgggtgg
     5641 aagagatgaa ggttacgatt ggttgattat gacacccggt gtgggtttag atgacaaggg
     5701 agacgcattg ggtcaacagt atagaaccgt ggatgatgtg gtctctacag gatctgacat
     5761 tattattgtt ggaagaggac tatttgcaaa gggaagggat gctaaggtag agggtgaacg
     5821 ttacagaaaa gcaggctggg aagcatattt gagaagatgc ggccagcaaa actaaaaaac
     5881 tgtattataa gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa tttaattata
     5941 tcagttatta ccctgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     6001 aggaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     6061 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     6121 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     6181 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     6241 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     6301 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     6361 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     6421 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc gcgccattcg ccattcaggc
     6481 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     6541 aggggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     6601 gttgtaaaac gacggccagt gaattgtaat acgactcact atagggcgaa ttggagct
//
