LOCUS       40924_7256        6613 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Yeast integrative vector pBS46, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6613)
  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  JOURNAL   Unpublished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 6613)
  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 6613)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6613)
  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            "Unpublished"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..6613
                     /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(2079..2095)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      2144..2333
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(2356..2374)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2395..2411)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2419..2435)
                     /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(2443..2473)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2488..2509)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2797..3385)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3559..4416)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4417..4521)
                     /label=AmpR promoter
     promoter        4806..5026
                     /label=URA3 promoter
     CDS             5027..5827
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(5959..6414)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     6559..6575
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6582..6600
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        6613
                     /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 ccacctccac ctcccggcgc
     1261 cagtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga
     1321 tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa
     1381 acttaccctt aaatttattt gcactactgg aaaactacct gttccatggc caacacttgt
     1441 cactactttg acttggggtg ttcaatgctt ttcaagatac ccagatcata tgaaacggca
     1501 tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaaagaacta tatttttcaa
     1561 agatgacggg aactacaaga cacgtgctga agtcaagttt gaaggtgata cccttgttaa
     1621 tagaatcgag ttaaaaggta ttgattttaa agaagatgga aacattcttg gacacaaatt
     1681 ggaatacaac tatatttcac acaatgtata catcactgca gacaaacaaa agaatggaat
     1741 caaagctcat ttcaaaatta gacacaacat tgaagatgga agcgttcaac tagcagacca
     1801 ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct
     1861 gtccacacaa tctgcccttt cgaaagatcc caacgaaaag agagaccaca tggtccttct
     1921 tgagtttgta acagctgctg ggattacaca tggcatggat gaactataca aatagggcgc
     1981 gccacttcta aataagcgaa tttcttatga tttataaggg cgaattccag cacactggcg
     2041 gccgttacta gtggatccga gctcggtacc aagcttatcg ataccgtcga cctcgagtca
     2101 tgtaattagt tatgtcacgc ttacattcac gccctccccc cacatccgct ctaaccgaaa
     2161 aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata gttatgttag
     2221 tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg
     2281 catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg aaggcttggt
     2341 acccagcttt tgttcccttt agtgagggtt aattccgagc ttggcgtaat catggtcata
     2401 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatag gagccggaag
     2461 cataaagtgt aaagcctggg gtgcctaatg agtgaggtaa ctcacattaa ttgcgttgcg
     2521 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     2581 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2641 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2701 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2761 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcg gcccccctga
     2821 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2881 ataccaggcg ttcccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2941 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     3001 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3061 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3121 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3181 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     3241 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3301 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3361 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3421 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3481 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3541 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3601 atttcgttca tccatagttg cctgactgcc cgtcgtgtag ataactacga tacgggaggg
     3661 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3721 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3781 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3841 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3901 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3961 gttgtgaaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4021 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4081 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4141 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4201 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4261 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4321 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4381 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4441 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4501 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     4561 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     4621 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     4681 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     4741 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     4801 cgcttttcaa ttcaattcat catttttttt ttattctttt ttttgatttc ggtttctttg
     4861 aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact
     4921 tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt
     4981 aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaatcatgt cgaaagctac
     5041 atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat ttaatatcat
     5101 gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca aggaattact
     5161 ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg tggatatctt
     5221 gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg ccaagtacaa
     5281 ttttttactc ttcgaagaca gaaaatttgc tgacattggt aatacagtca aattgcagta
     5341 ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac acggtgtggt
     5401 gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa aggaacctag
     5461 aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg gagaatatac
     5521 taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct ttattgctca
     5581 aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac ccggtgtggg
     5641 tttagatgac aagggagacg cattgggtca acagtataga accgtggatg atgtggtctc
     5701 tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa gggatgctaa
     5761 ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa gatgcggcca
     5821 gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc
     5881 ttcaatttaa ttatatcagt tattaccctg cggtgtgaaa taccgcacag atgcgtaagg
     5941 agaaaatacc gcatcaggaa attgtaaacg ttaatatttt gttaaaattc gcgttaaatt
     6001 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     6061 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     6121 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     6181 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     6241 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     6301 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     6361 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcgcgcc
     6421 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     6481 tacgccagct ggcgaagggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     6541 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac tcactatagg
     6601 gcgaattgga gct
//
