LOCUS       40924_7236        6592 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Yeast integrative vector pBS42, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6592)
  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  JOURNAL   Unpublished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 6592)
  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 6592)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6592)
  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            "Unpublished"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..6592
                     /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(2058..2074)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      2123..2312
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(2335..2353)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2374..2390)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2398..2414)
                     /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(2422..2452)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2467..2488)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2776..3364)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3538..4395)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4396..4500)
                     /label=AmpR promoter
     promoter        4785..5005
                     /label=URA3 promoter
     CDS             5006..5806
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(5938..6393)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     6538..6554
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6561..6579
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        6592
                     /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 agtaaaggag aagaactttt
     1261 cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc acaaattttc
     1321 tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa cttaccctta aatttatttg
     1381 cactactgga aaactacctg ttccatggcc aacacttgtc actactttga cttggggtgt
     1441 tcaatgcttt tcaagatacc cagatcatat gaaacggcat gactttttca agagtgccat
     1501 gcccgaaggt tatgtacagg aaagaactat atttttcaaa gatgacggga actacaagac
     1561 acgtgctgaa gtcaagtttg aaggtgatac ccttgttaat agaatcgagt taaaaggtat
     1621 tgattttaaa gaagatggaa acattcttgg acacaaattg gaatacaact atatttcaca
     1681 caatgtatac atcactgcag acaaacaaaa gaatggaatc aaagctcatt tcaaaattag
     1741 acacaacatt gaagatggaa gcgttcaact agcagaccat tatcaacaaa atactccaat
     1801 tggcgatggc cctgtccttt taccagacaa ccattacctg tccacacaat ctgccctttc
     1861 gaaagatccc aacgaaaaga gagaccacat ggtccttctt gagtttgtaa cagctgctgg
     1921 gattacacat ggcatggatg aactatacaa atagggcgcg ccacttctaa ataagcgaat
     1981 ttcttatgat ttataagggc gaattccagc acactggcgg ccgttactag tggatccgag
     2041 ctcggtacca agcttatcga taccgtcgac ctcgagtcat gtaattagtt atgtcacgct
     2101 tacattcacg ccctcccccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg
     2161 aagtctaggt ccctatttat ttttttatag ttatgttagt attaagaacg ttatttatat
     2221 ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa
     2281 ccttgcttga gaaggttttg ggacgctcga aggcttggta cccagctttt gttcccttta
     2341 gtgagggtta attccgagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     2401 ttatccgctc acaattccac acaacatagg agccggaagc ataaagtgta aagcctgggg
     2461 tgcctaatga gtgaggtaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     2521 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     2581 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     2641 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2701 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2761 cgcgttgctg gcgtttttcc ataggctcgg cccccctgac gagcatcaca aaaatcgacg
     2821 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt tcccccctgg
     2881 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2941 tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt
     3001 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3061 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3121 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3181 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     3241 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3301 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3361 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3421 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3481 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3541 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3601 ctgactgccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     3661 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3721 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3781 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3841 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3901 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgaaaaa aagcggttag
     3961 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4021 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4081 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4141 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4201 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4261 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4321 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4381 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4441 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4501 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     4561 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     4621 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     4681 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     4741 ctatgcggca tcagagcaga ttgtactgag agtgcaccac gcttttcaat tcaattcatc
     4801 attttttttt tattcttttt tttgatttcg gtttctttga aatttttttg attcggtaat
     4861 ctccgaacag aaggaagaac gaaggaagga gcacagactt agattggtat atatacgcat
     4921 atgtagtgtt gaagaaacat gaaattgccc agtattctta acccaactgc acagaacaaa
     4981 aacctgcagg aaacgaagat aaatcatgtc gaaagctaca tataaggaac gtgctgctac
     5041 tcatcctagt cctgttgctg ccaagctatt taatatcatg cacgaaaagc aaacaaactt
     5101 gtgtgcttca ttggatgttc gtaccaccaa ggaattactg gagttagttg aagcattagg
     5161 tcccaaaatt tgtttactaa aaacacatgt ggatatcttg actgattttt ccatggaggg
     5221 cacagttaag ccgctaaagg cattatccgc caagtacaat tttttactct tcgaagacag
     5281 aaaatttgct gacattggta atacagtcaa attgcagtac tctgcgggtg tatacagaat
     5341 agcagaatgg gcagacatta cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg
     5401 tttgaagcag gcggcagaag aagtaacaaa ggaacctaga ggccttttga tgttagcaga
     5461 attgtcatgc aagggctccc tatctactgg agaatatact aagggtactg ttgacattgc
     5521 gaagagcgac aaagattttg ttatcggctt tattgctcaa agagacatgg gtggaagaga
     5581 tgaaggttac gattggttga ttatgacacc cggtgtgggt ttagatgaca agggagacgc
     5641 attgggtcaa cagtatagaa ccgtggatga tgtggtctct acaggatctg acattattat
     5701 tgttggaaga ggactatttg caaagggaag ggatgctaag gtagagggtg aacgttacag
     5761 aaaagcaggc tgggaagcat atttgagaag atgcggccag caaaactaaa aaactgtatt
     5821 ataagtaaat gcatgtatac taaactcaca aattagagct tcaatttaat tatatcagtt
     5881 attaccctgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa
     5941 ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     6001 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     6061 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     6121 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     6181 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     6241 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     6301 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     6361 ccgccgcgct taatgcgccg ctacagggcg cgtcgcgcca ttcgccattc aggctgcgca
     6421 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaggggg
     6481 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     6541 aaacgacggc cagtgaattg taatacgact cactataggg cgaattggag ct
//
