LOCUS       40924_7246        5869 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Yeast integrative vector pBS43, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5869)
  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  JOURNAL   Unpublished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 5869)
  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 5869)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5869)
  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            "Unpublished"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..5869
                     /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..1228
                     /codon_start=1
                     /label=EYFP
                     /note="enhanced YFP"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(1335..1351)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      1400..1589
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(1612..1630)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1651..1667)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1675..1691)
                     /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(1699..1729)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1744..1765)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2053..2641)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2815..3672)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3673..3777)
                     /label=AmpR promoter
     promoter        4062..4282
                     /label=URA3 promoter
     CDS             4283..5083
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(5215..5670)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     5815..5831
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5838..5856
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        5869
                     /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 aattaacagt aaaggagaag aacttttcac
      541 tggagttgtc ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt
      601 cagtggagag ggtgaaggtg atgcaacata cggaaaactt acccttaaat ttatttgcac
      661 tactggaaaa ctacctgttc catggccaac acttgtcact actttcggtt atggtctaaa
      721 atgctttgct agatacccag atcatatgaa acggcatgac tttttcaaga gtgccatgcc
      781 cgaaggttat gtacaggaaa gaactatatt tttcaaagat gacgggaact acaagacacg
      841 tgctgaagtc aagtttgaag gtgataccct tgttaataga atcgagttaa aaggtattga
      901 ttttaaagaa gatggaaaca ttcttggaca caaattggaa tacaactata actcacacaa
      961 tgtatacatc atggcagaca aacaaaagaa tggaatcaaa gttaacttca aaattagaca
     1021 caacattgaa gatggaagcg ttcaactagc agaccattat caacaaaata ctccaattgg
     1081 cgatggccct gtccttttac cagacaacca ttacctgtcc tatcaatctg ccctttcgaa
     1141 agatcccaac gaaaagagag accacatggt ccttcttgag tttgtaacag ctgctgggat
     1201 tacacatggc atggatgaac tatacaaata gggcgcgcca cttctaaata agcgaatttc
     1261 ttatgattta taagggcgaa ttccagcaca ctggcggccg ttactagtgg atccgagctc
     1321 ggtaccaagc ttatcgatac cgtcgacctc gagtcatgta attagttatg tcacgcttac
     1381 attcacgccc tccccccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag
     1441 tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta tttatatttc
     1501 aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct
     1561 tgcttgagaa ggttttggga cgctcgaagg cttggtaccc agcttttgtt ccctttagtg
     1621 agggttaatt ccgagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     1681 tccgctcaca attccacaca acataggagc cggaagcata aagtgtaaag cctggggtgc
     1741 ctaatgagtg aggtaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     1801 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     1861 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     1921 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     1981 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2041 gttgctggcg tttttccata ggctcggccc ccctgacgag catcacaaaa atcgacgctc
     2101 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgttcc cccctggaag
     2161 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2221 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     2281 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2341 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2401 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2461 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     2521 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2581 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2641 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     2701 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     2761 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     2821 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     2881 actgcccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     2941 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3001 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3061 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3121 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3181 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgaaaaaaag cggttagctc
     3241 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     3301 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     3361 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     3421 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     3481 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     3541 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     3601 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     3661 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     3721 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     3781 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     3841 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     3901 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     3961 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     4021 tgcggcatca gagcagattg tactgagagt gcaccacgct tttcaattca attcatcatt
     4081 ttttttttat tctttttttt gatttcggtt tctttgaaat ttttttgatt cggtaatctc
     4141 cgaacagaag gaagaacgaa ggaaggagca cagacttaga ttggtatata tacgcatatg
     4201 tagtgttgaa gaaacatgaa attgcccagt attcttaacc caactgcaca gaacaaaaac
     4261 ctgcaggaaa cgaagataaa tcatgtcgaa agctacatat aaggaacgtg ctgctactca
     4321 tcctagtcct gttgctgcca agctatttaa tatcatgcac gaaaagcaaa caaacttgtg
     4381 tgcttcattg gatgttcgta ccaccaagga attactggag ttagttgaag cattaggtcc
     4441 caaaatttgt ttactaaaaa cacatgtgga tatcttgact gatttttcca tggagggcac
     4501 agttaagccg ctaaaggcat tatccgccaa gtacaatttt ttactcttcg aagacagaaa
     4561 atttgctgac attggtaata cagtcaaatt gcagtactct gcgggtgtat acagaatagc
     4621 agaatgggca gacattacga atgcacacgg tgtggtgggc ccaggtattg ttagcggttt
     4681 gaagcaggcg gcagaagaag taacaaagga acctagaggc cttttgatgt tagcagaatt
     4741 gtcatgcaag ggctccctat ctactggaga atatactaag ggtactgttg acattgcgaa
     4801 gagcgacaaa gattttgtta tcggctttat tgctcaaaga gacatgggtg gaagagatga
     4861 aggttacgat tggttgatta tgacacccgg tgtgggttta gatgacaagg gagacgcatt
     4921 gggtcaacag tatagaaccg tggatgatgt ggtctctaca ggatctgaca ttattattgt
     4981 tggaagagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac gttacagaaa
     5041 agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa ctgtattata
     5101 agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat atcagttatt
     5161 accctgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggaaattg
     5221 taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     5281 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     5341 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     5401 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     5461 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     5521 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     5581 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     5641 ccgcgcttaa tgcgccgcta cagggcgcgt cgcgccattc gccattcagg ctgcgcaact
     5701 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaggggggat
     5761 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     5821 cgacggccag tgaattgtaa tacgactcac tatagggcga attggagct
//
