LOCUS       40924_7241        6575 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Yeast integrative vector pBS42BN, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6575)
  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  JOURNAL   Unpublished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 6575)
  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 6575)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6575)
  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            "Unpublished"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..6575
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        34..430
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             466..486
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             496..516
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             520..540
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             550..1266
                     /codon_start=1
                     /label=EYFP
                     /note="enhanced YFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     misc_feature    1270..1293
                     /label=BamHI and NheI sites
                     /note="BamHI and NheI sites"
     terminator      2138..2327
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(2350..2368)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2389..2405)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2413..2429)
                     /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(2437..2467)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2482..2503)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2791..3379)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3553..4410)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4411..4515)
                     /label=AmpR promoter
     promoter        4800..5020
                     /label=URA3 promoter
     CDS             5021..5821
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(5953..6408)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     6553..6569
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg cgaattggag ctccagcttc aaaatgtttc tactcctttt
       61 ttactcttcc agattttctc ggactccgcg catcgccgta ccacttcaaa acacccaagc
      121 acagcatact aaatttcccc tctttcttcc tctagggtgt cgttaattac ccgtactaaa
      181 ggtttggaaa agaaaaaaga gaccgcctcg tttctttttc ttcgtcgaaa aaggcaataa
      241 aaatttttat cacgtttctt tttcttgaaa attttttttt tgattttttt ctctttcgat
      301 gacctcccat tgatatttaa gttaataaac ggtcttcaat ttctcaagtt tcagtttcat
      361 ttttcttgtt ctattacaac tttttttact tcttgctcat tagaaagaaa gcatagcaat
      421 ctaatctaag ttttctagat atctgcagaa ttcgccctta tggttccaaa gaagaagaga
      481 aaggttttaa ttgatccaaa aaagaaaaga aaggttgatc caaagaaaaa gagaaaagtt
      541 ttaattaaca tggtgagtaa aggagaagaa cttttcactg gagttgtccc aattcttgtt
      601 gaattagatg gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat
      661 gcaacatacg gaaaacttac ccttaaattt atttgcacta ctggaaaact acctgttcca
      721 tggccaacac ttgtcactac tttcggttat ggtctaaaat gctttgctag atacccagat
      781 catatgaaac ggcatgactt tttcaagagt gccatgcccg aaggttatgt acaggaaaga
      841 actatatttt tcaaagatga cgggaactac aagacacgtg ctgaagtcaa gtttgaaggt
      901 gatacccttg ttaatagaat cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt
      961 cttggacaca aattggaata caactataac tcacacaatg tatacatcat ggcagacaaa
     1021 caaaagaatg gaatcaaagt taacttcaaa attagacaca acattgaaga tggaagcgtt
     1081 caactagcag accattatca acaaaatact ccaattggcg atggccctgt ccttttacca
     1141 gacaaccatt acctgtccta tcaatctgcc ctttcgaaag atcccaacga aaagagagac
     1201 cacatggtcc ttcttgagtt tgtaacagct gctgggatta cacatggcat ggatgaacta
     1261 tacaaaggcg gatccgcagc cggcgcagct agcgccagta aaggagaaga acttttcact
     1321 ggagttgtcc caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc
     1381 agtggagagg gtgaaggtga tgcaacatac ggaaaactta cccttaaatt tatttgcact
     1441 actggaaaac tacctgttcc atggccaaca cttgtcacta ctttgacttg gggtgttcaa
     1501 tgcttttcaa gatacccaga tcatatgaaa cggcatgact ttttcaagag tgccatgccc
     1561 gaaggttatg tacaggaaag aactatattt ttcaaagatg acgggaacta caagacacgt
     1621 gctgaagtca agtttgaagg tgataccctt gttaatagaa tcgagttaaa aggtattgat
     1681 tttaaagaag atggaaacat tcttggacac aaattggaat acaactatat ttcacacaat
     1741 gtatacatca ctgcagacaa acaaaagaat ggaatcaaag ctcatttcaa aattagacac
     1801 aacattgaag atggaagcgt tcaactagca gaccattatc aacaaaatac tccaattggc
     1861 gatggccctg tccttttacc agacaaccat tacctgtcca cacaatctgc cctttcgaaa
     1921 gatcccaacg aaaagagaga ccacatggtc cttcttgagt ttgtaacagc tgctgggatt
     1981 acacatggca tggatgaact atacaaatag ggcgcgccac ttctaaataa gcgaatttct
     2041 tatgatttat aagggcgaat tccagcacac tggcggccgt tactagtcga gtcatgtaat
     2101 tagttatgtc acgcttacat tcacgccctc cccccacatc cgctctaacc gaaaaggaag
     2161 gagttagaca acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa
     2221 gaacgttatt tatatttcaa atttttcttt tttttctgta cagacgcgtg tacgcatgta
     2281 acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct tggtacccag
     2341 cttttgttcc ctttagtgag ggttaattcc gagcttggcg taatcatggt catagctgtt
     2401 tcctgtgtga aattgttatc cgctcacaat tccacacaac ataggagccg gaagcataaa
     2461 gtgtaaagcc tggggtgcct aatgagtgag gtaactcaca ttaattgcgt tgcgctcact
     2521 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     2581 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     2641 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2701 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2761 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctcggccccc ctgacgagca
     2821 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2881 ggcgttcccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2941 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     3001 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3061 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3121 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3181 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     3241 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3301 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3361 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3421 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3481 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3541 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3601 ttcatccata gttgcctgac tgcccgtcgt gtagataact acgatacggg agggcttacc
     3661 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3721 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3781 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3841 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     3901 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     3961 aaaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4021 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4081 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4141 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4201 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4261 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4321 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4381 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4441 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4501 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     4561 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     4621 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     4681 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     4741 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accacgcttt
     4801 tcaattcaat tcatcatttt ttttttattc ttttttttga tttcggtttc tttgaaattt
     4861 ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca gacttagatt
     4921 ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat tcttaaccca
     4981 actgcacaga acaaaaacct gcaggaaacg aagataaatc atgtcgaaag ctacatataa
     5041 ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata tcatgcacga
     5101 aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat tactggagtt
     5161 agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata tcttgactga
     5221 tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt acaatttttt
     5281 actcttcgaa gacagaaaat ttgctgacat tggtaataca gtcaaattgc agtactctgc
     5341 gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg tggtgggccc
     5401 aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac ctagaggcct
     5461 tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat atactaaggg
     5521 tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg ctcaaagaga
     5581 catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg tgggtttaga
     5641 tgacaaggga gacgcattgg gtcaacagta tagaaccgtg gatgatgtgg tctctacagg
     5701 atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg ctaaggtaga
     5761 gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg gccagcaaaa
     5821 ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta gagcttcaat
     5881 ttaattatat cagttattac cctgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     5941 taccgcatca ggaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt
     6001 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     6061 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     6121 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     6181 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     6241 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     6301 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     6361 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcg cgccattcgc
     6421 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     6481 agctggcgaa ggggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     6541 agtcacgacg ttgtaaaacg acggccagtg aattg
//
