LOCUS       40924_7271        7000 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Yeast integrative vector pBS49, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7000)
  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  JOURNAL   Unpublished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 7000)
  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 7000)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7000)
  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            "Unpublished"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..7000
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     128..144
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        151..169
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        184..580
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             616..636
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             646..666
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             670..690
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             700..1410
                     /codon_start=1
                     /label=EYFP
                     /note="enhanced YFP"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(1517..1533)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      1582..1771
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(1794..1812)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1833..1849)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1857..1873)
                     /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(1881..1911)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1926..1947)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2235..2823)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2997..3854)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3855..3959)
                     /label=AmpR promoter
     CDS             complement(4720..5811)
                     /codon_start=1
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(5824..6228)
                     /label=LEU2 promoter
     rep_origin      complement(6528..6983)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
       61 gcgaaggggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      121 cgacgttgta aaacgacggc cagtgaattg taatacgact cactataggg cgaattggag
      181 ctccagcttc aaaatgtttc tactcctttt ttactcttcc agattttctc ggactccgcg
      241 catcgccgta ccacttcaaa acacccaagc acagcatact aaatttcccc tctttcttcc
      301 tctagggtgt cgttaattac ccgtactaaa ggtttggaaa agaaaaaaga gaccgcctcg
      361 tttctttttc ttcgtcgaaa aaggcaataa aaatttttat cacgtttctt tttcttgaaa
      421 attttttttt tgattttttt ctctttcgat gacctcccat tgatatttaa gttaataaac
      481 ggtcttcaat ttctcaagtt tcagtttcat ttttcttgtt ctattacaac tttttttact
      541 tcttgctcat tagaaagaaa gcatagcaat ctaatctaag ttttctagat atctgcagaa
      601 ttcgccctta tggttccaaa gaagaagaga aaggttttaa ttgatccaaa aaagaaaaga
      661 aaggttgatc caaagaaaaa gagaaaagtt ttaattaaca gtaaaggaga agaacttttc
      721 actggagttg tcccaattct tgttgaatta gatggtgatg ttaatgggca caaattttct
      781 gtcagtggag agggtgaagg tgatgcaaca tacggaaaac ttacccttaa atttatttgc
      841 actactggaa aactacctgt tccatggcca acacttgtca ctactttcgg ttatggtcta
      901 aaatgctttg ctagataccc agatcatatg aaacggcatg actttttcaa gagtgccatg
      961 cccgaaggtt atgtacagga aagaactata tttttcaaag atgacgggaa ctacaagaca
     1021 cgtgctgaag tcaagtttga aggtgatacc cttgttaata gaatcgagtt aaaaggtatt
     1081 gattttaaag aagatggaaa cattcttgga cacaaattgg aatacaacta taactcacac
     1141 aatgtataca tcatggcaga caaacaaaag aatggaatca aagttaactt caaaattaga
     1201 cacaacattg aagatggaag cgttcaacta gcagaccatt atcaacaaaa tactccaatt
     1261 ggcgatggcc ctgtcctttt accagacaac cattacctgt cctatcaatc tgccctttcg
     1321 aaagatccca acgaaaagag agaccacatg gtccttcttg agtttgtaac agctgctggg
     1381 attacacatg gcatggatga actatacaaa tagggcgcgc cacttctaaa taagcgaatt
     1441 tcttatgatt tataagggcg aattccagca cactggcggc cgttactagt ggatccgagc
     1501 tcggtaccaa gcttatcgat accgtcgacc tcgagtcatg taattagtta tgtcacgctt
     1561 acattcacgc cctcccccca catccgctct aaccgaaaag gaaggagtta gacaacctga
     1621 agtctaggtc cctatttatt tttttatagt tatgttagta ttaagaacgt tatttatatt
     1681 tcaaattttt cttttttttc tgtacagacg cgtgtacgca tgtaacatta tactgaaaac
     1741 cttgcttgag aaggttttgg gacgctcgaa ggcttggtac ccagcttttg ttccctttag
     1801 tgagggttaa ttccgagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     1861 tatccgctca caattccaca caacatagga gccggaagca taaagtgtaa agcctggggt
     1921 gcctaatgag tgaggtaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     1981 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     2041 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     2101 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     2161 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2221 gcgttgctgg cgtttttcca taggctcggc ccccctgacg agcatcacaa aaatcgacgc
     2281 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt cccccctgga
     2341 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2401 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     2461 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2521 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2581 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2641 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     2701 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2761 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2821 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2881 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2941 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     3001 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     3061 tgactgcccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     3121 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3181 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3241 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3301 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     3361 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     3421 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     3481 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     3541 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     3601 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     3661 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     3721 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     3781 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     3841 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     3901 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     3961 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     4021 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     4081 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     4141 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     4201 tatgcggcat cagagcagat tgtactgaga gtgcaccata tcgactacgt cgtaaggccg
     4261 tttctgacag agtaaaattc ttgagggaac tttcaccatt atgggaaatg gttcaagaag
     4321 gtattgactt aaactccatc aaatggtcag gtcattgagt gttttttatt tgttgtattt
     4381 tttttttttt agagaaaatc ctccaatatc aaattaggaa tcgtagtttc atgattttct
     4441 gttacaccta actttttgtg tggtgccctc ctccttgtca atattaatgt taaagtgcaa
     4501 ttctttttcc ttatcacgtt gagccattag tatcaatttg cttacctgta ttcctttact
     4561 atcctccttt ttctccttct tgataaatgt atgtagattg cgtatatagt ttcgtctacc
     4621 ctatgaacat attccatttt gtaatttcgt gtcgtttcta ttatgaattt catttataaa
     4681 gtttatgtac aaatatcata aaaaaagaga atctttttaa gcaaggattt tcttaacttc
     4741 ttcggcgaca gcatcaccga cttcggtggt actgttggaa ccacctaaat caccagttct
     4801 gatacctgca tccaaaacct ttttaactgc atcttcaatg gccttacctt cttcaggcaa
     4861 gttcaatgac aatttcaaca tcattgcagc agacaagata gtggcgatag ggtcaacctt
     4921 attctttggc aaatctggag cagaaccgtg gcatggttcg tacaaaccaa atgcggtgtt
     4981 cttgtctggc aaagaggcca aggacgcaga tggcaacaaa cccaaggaac ctgggataac
     5041 ggaggcttca tcggagatga tatcaccaaa catgttgctg gtgattataa taccatttag
     5101 gtgggttggg ttcttaacta ggatcatggc ggcagaatca atcaattgat gttgaacctt
     5161 caatgtaggg aattcgttct tgatggtttc ctccacagtt tttctccata atcttgaaga
     5221 ggccaaaaca ttagctttat ccaaggacca aataggcaat ggtggctcat gttgtagggc
     5281 catgaaagcg gccattcttg tgattctttg cacttctgga acggtgtatt gttcactatc
     5341 ccaagcgaca ccatcaccat cgtcttcctt tctcttacca aagtaaatac ctcccactaa
     5401 ttctctgaca acaacgaagt cagtaccttt agcaaattgt ggcttgattg gagataagtc
     5461 taaaagagag tcggatgcaa agttacatgg tcttaagttg gcgtacaatt gaagttcttt
     5521 acggattttt agtaaacctt gttcaggtct aacactaccg gtaccccatt taggaccacc
     5581 cacagcacct aacaaaacgg catcaacctt cttggaggct tccagcgcct catctggaag
     5641 tgggacacct gtagcatcga tagcagcacc accaattaaa tgattttcga aatcgaactt
     5701 gacattggaa cgaacatcag aaatagcttt aagaacctta atggcttcgg ctgtgatttc
     5761 ttgaccaacg tggtcacctg gcaaaacgac gatcttctta ggggcagaca taggggcaga
     5821 cattagaatg gtatatcctt gaaatatata tatatattgc tgaaatgtaa aaggtaagaa
     5881 aagttagaaa gtaagacgat tgctaaccac ctattggaaa aaacaatagg tccttaaata
     5941 atattgtcaa cttcaagtat tgtgatgcaa gcatttagtc atgaacgctt ctctattcta
     6001 tatgaaaagc cggttccggc ctctcacctt tcctttttct cccaattttt cagttgaaaa
     6061 aggtatatgc gtcaggcgac ctctgaaatt aacaaaaaat ttccagtcat cgaatttgat
     6121 tctgtgcgat agcgcccctg tgtgttctcg ttatgttgag gaaaaaaata atggttgcta
     6181 agagattcga actcttgcat cttacgatac ctgagtattc ccacagttaa ctgcggtcaa
     6241 gatatttctt gaatcaggcg ccttagaccg ctcggccaaa caaccaatta cttgttgaga
     6301 aatagagtat aattatccta taaatataac gtttttgaac acacatgaac aaggaagtac
     6361 aggacaattg attttgaaga gaatgtggat tttgatgtaa ttgttgggat tccattttta
     6421 ataaggcaat aatattaggt atgtggatat actagaagtt ctcctcgacc gtcgatatgc
     6481 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa ttgtaaacgt
     6541 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     6601 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     6661 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     6721 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     6781 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     6841 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     6901 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     6961 taatgcgccg ctacagggcg cgtcgcgcca ttcgccattc
//
