LOCUS       40924_7266        6643 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Yeast integrative vector pBS48, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6643)
  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  JOURNAL   Unpublished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 6643)
  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 6643)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6643)
  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            "Unpublished"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..6643
                     /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(2109..2125)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      2174..2363
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(2386..2404)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2425..2441)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2449..2465)
                     /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(2473..2503)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2518..2539)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2827..3415)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3589..4446)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4447..4551)
                     /label=AmpR promoter
     promoter        4836..5056
                     /label=URA3 promoter
     CDS             5057..5857
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(5989..6444)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     6589..6605
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6612..6630
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        6643
                     /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 accacctcca cctccacctc cacccggcgc cagtaaagga gaagaacttt tcactggagt
     1321 tgtcccaatt cttgttgaat tagatggtga tgttaatggg cacaaatttt ctgtcagtgg
     1381 agagggtgaa ggtgatgcaa catacggaaa acttaccctt aaatttattt gcactactgg
     1441 aaaactacct gttccatggc caacacttgt cactactttg acttggggtg ttcaatgctt
     1501 ttcaagatac ccagatcata tgaaacggca tgactttttc aagagtgcca tgcccgaagg
     1561 ttatgtacag gaaagaacta tatttttcaa agatgacggg aactacaaga cacgtgctga
     1621 agtcaagttt gaaggtgata cccttgttaa tagaatcgag ttaaaaggta ttgattttaa
     1681 agaagatgga aacattcttg gacacaaatt ggaatacaac tatatttcac acaatgtata
     1741 catcactgca gacaaacaaa agaatggaat caaagctcat ttcaaaatta gacacaacat
     1801 tgaagatgga agcgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg
     1861 ccctgtcctt ttaccagaca accattacct gtccacacaa tctgcccttt cgaaagatcc
     1921 caacgaaaag agagaccaca tggtccttct tgagtttgta acagctgctg ggattacaca
     1981 tggcatggat gaactataca aatagggcgc gccacttcta aataagcgaa tttcttatga
     2041 tttataaggg cgaattccag cacactggcg gccgttacta gtggatccga gctcggtacc
     2101 aagcttatcg ataccgtcga cctcgagtca tgtaattagt tatgtcacgc ttacattcac
     2161 gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg
     2221 tccctattta tttttttata gttatgttag tattaagaac gttatttata tttcaaattt
     2281 ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa accttgcttg
     2341 agaaggtttt gggacgctcg aaggcttggt acccagcttt tgttcccttt agtgagggtt
     2401 aattccgagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     2461 cacaattcca cacaacatag gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     2521 agtgaggtaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2581 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2641 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2701 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2761 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2821 ggcgtttttc cataggctcg gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2881 gaggtggcga aacccgacag gactataaag ataccaggcg ttcccccctg gaagctccct
     2941 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3001 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3061 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3121 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3181 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3241 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     3301 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3361 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3421 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3481 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3541 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3601 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactgcc
     3661 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3721 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3781 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3841 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3901 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3961 acgatcaagg cgagttacat gatcccccat gttgtgaaaa aaagcggtta gctccttcgg
     4021 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4081 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4141 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4201 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4261 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4321 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4381 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4441 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4501 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4561 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     4621 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     4681 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     4741 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     4801 atcagagcag attgtactga gagtgcacca cgcttttcaa ttcaattcat catttttttt
     4861 ttattctttt ttttgatttc ggtttctttg aaattttttt gattcggtaa tctccgaaca
     4921 gaaggaagaa cgaaggaagg agcacagact tagattggta tatatacgca tatgtagtgt
     4981 tgaagaaaca tgaaattgcc cagtattctt aacccaactg cacagaacaa aaacctgcag
     5041 gaaacgaaga taaatcatgt cgaaagctac atataaggaa cgtgctgcta ctcatcctag
     5101 tcctgttgct gccaagctat ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc
     5161 attggatgtt cgtaccacca aggaattact ggagttagtt gaagcattag gtcccaaaat
     5221 ttgtttacta aaaacacatg tggatatctt gactgatttt tccatggagg gcacagttaa
     5281 gccgctaaag gcattatccg ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc
     5341 tgacattggt aatacagtca aattgcagta ctctgcgggt gtatacagaa tagcagaatg
     5401 ggcagacatt acgaatgcac acggtgtggt gggcccaggt attgttagcg gtttgaagca
     5461 ggcggcagaa gaagtaacaa aggaacctag aggccttttg atgttagcag aattgtcatg
     5521 caagggctcc ctatctactg gagaatatac taagggtact gttgacattg cgaagagcga
     5581 caaagatttt gttatcggct ttattgctca aagagacatg ggtggaagag atgaaggtta
     5641 cgattggttg attatgacac ccggtgtggg tttagatgac aagggagacg cattgggtca
     5701 acagtataga accgtggatg atgtggtctc tacaggatct gacattatta ttgttggaag
     5761 aggactattt gcaaagggaa gggatgctaa ggtagagggt gaacgttaca gaaaagcagg
     5821 ctgggaagca tatttgagaa gatgcggcca gcaaaactaa aaaactgtat tataagtaaa
     5881 tgcatgtata ctaaactcac aaattagagc ttcaatttaa ttatatcagt tattaccctg
     5941 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaaacg
     6001 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     6061 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     6121 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     6181 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     6241 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     6301 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     6361 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     6421 ttaatgcgcc gctacagggc gcgtcgcgcc attcgccatt caggctgcgc aactgttggg
     6481 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaagggg ggatgtgctg
     6541 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     6601 ccagtgaatt gtaatacgac tcactatagg gcgaattgga gct
//
