LOCUS       40924_42529        6476 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast integrative vector pTAF18, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6476)
  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  JOURNAL   Unpublished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 6476)
  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 6476)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6476)
  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            "Unpublished"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..6476
                     /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"
     primer_bind     complement(843..859)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      908..1097
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(1120..1138)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1159..1175)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1183..1199)
                     /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(1207..1237)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1252..1273)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1561..2149)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2323..3180)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3181..3285)
                     /label=AmpR promoter
     CDS             complement(4046..5137)
                     /codon_start=1
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(5150..5554)
                     /label=LEU2 promoter
     rep_origin      complement(5854..6309)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     6454..6470
                     /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 ttaattaacc ttttttctaa agaattgcga tgtatgatgt atggctttgg ggatgaccag
      601 aatccttata ctgagtcagt ggatattctt gaagatcttg tcatagagtt tatcactgaa
      661 atgactcaca aggcaatgtc aattggaaga caaggtcgag tacaagttga agatatcgtc
      721 ttcttgattc gaaaggaccc aaggaagttt gccagggtta aagacttgct tactatgaat
      781 gaagaattga aacgagctag aaaagcattt gatgaagcaa attatggatc ttagaagctt
      841 atcgataccg tcgacctcga gtcatgtaat tagttatgtc acgcttacat tcacgccctc
      901 cccccacatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta
      961 tttatttttt tatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt
     1021 tttttctgta cagacgcgtg tacgcatgta acattatact gaaaaccttg cttgagaagg
     1081 ttttgggacg ctcgaaggct tggtacccag cttttgttcc ctttagtgag ggttaattcc
     1141 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     1201 tccacacaac ataggagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     1261 gtaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     1321 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     1381 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     1441 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     1501 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     1561 tttccatagg ctcggccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     1621 gcgaaacccg acaggactat aaagatacca ggcgttcccc cctggaagct ccctcgtgcg
     1681 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     1741 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     1801 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     1861 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     1921 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     1981 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     2041 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2101 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2161 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2221 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2281 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2341 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tgcccgtcgt
     2401 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     2461 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     2521 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     2581 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     2641 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     2701 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     2761 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     2821 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     2881 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     2941 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     3001 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     3061 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     3121 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     3181 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     3241 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     3301 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     3361 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     3421 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     3481 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     3541 gcagattgta ctgagagtgc accatatcga ctacgtcgta aggccgtttc tgacagagta
     3601 aaattcttga gggaactttc accattatgg gaaatggttc aagaaggtat tgacttaaac
     3661 tccatcaaat ggtcaggtca ttgagtgttt tttatttgtt gtattttttt ttttttagag
     3721 aaaatcctcc aatatcaaat taggaatcgt agtttcatga ttttctgtta cacctaactt
     3781 tttgtgtggt gccctcctcc ttgtcaatat taatgttaaa gtgcaattct ttttccttat
     3841 cacgttgagc cattagtatc aatttgctta cctgtattcc tttactatcc tcctttttct
     3901 ccttcttgat aaatgtatgt agattgcgta tatagtttcg tctaccctat gaacatattc
     3961 cattttgtaa tttcgtgtcg tttctattat gaatttcatt tataaagttt atgtacaaat
     4021 atcataaaaa aagagaatct ttttaagcaa ggattttctt aacttcttcg gcgacagcat
     4081 caccgacttc ggtggtactg ttggaaccac ctaaatcacc agttctgata cctgcatcca
     4141 aaaccttttt aactgcatct tcaatggcct taccttcttc aggcaagttc aatgacaatt
     4201 tcaacatcat tgcagcagac aagatagtgg cgatagggtc aaccttattc tttggcaaat
     4261 ctggagcaga accgtggcat ggttcgtaca aaccaaatgc ggtgttcttg tctggcaaag
     4321 aggccaagga cgcagatggc aacaaaccca aggaacctgg gataacggag gcttcatcgg
     4381 agatgatatc accaaacatg ttgctggtga ttataatacc atttaggtgg gttgggttct
     4441 taactaggat catggcggca gaatcaatca attgatgttg aaccttcaat gtagggaatt
     4501 cgttcttgat ggtttcctcc acagtttttc tccataatct tgaagaggcc aaaacattag
     4561 ctttatccaa ggaccaaata ggcaatggtg gctcatgttg tagggccatg aaagcggcca
     4621 ttcttgtgat tctttgcact tctggaacgg tgtattgttc actatcccaa gcgacaccat
     4681 caccatcgtc ttcctttctc ttaccaaagt aaatacctcc cactaattct ctgacaacaa
     4741 cgaagtcagt acctttagca aattgtggct tgattggaga taagtctaaa agagagtcgg
     4801 atgcaaagtt acatggtctt aagttggcgt acaattgaag ttctttacgg atttttagta
     4861 aaccttgttc aggtctaaca ctaccggtac cccatttagg accacccaca gcacctaaca
     4921 aaacggcatc aaccttcttg gaggcttcca gcgcctcatc tggaagtggg acacctgtag
     4981 catcgatagc agcaccacca attaaatgat tttcgaaatc gaacttgaca ttggaacgaa
     5041 catcagaaat agctttaaga accttaatgg cttcggctgt gatttcttga ccaacgtggt
     5101 cacctggcaa aacgacgatc ttcttagggg cagacatagg ggcagacatt agaatggtat
     5161 atccttgaaa tatatatata tattgctgaa atgtaaaagg taagaaaagt tagaaagtaa
     5221 gacgattgct aaccacctat tggaaaaaac aataggtcct taaataatat tgtcaacttc
     5281 aagtattgtg atgcaagcat ttagtcatga acgcttctct attctatatg aaaagccggt
     5341 tccggcctct cacctttcct ttttctccca atttttcagt tgaaaaaggt atatgcgtca
     5401 ggcgacctct gaaattaaca aaaaatttcc agtcatcgaa tttgattctg tgcgatagcg
     5461 cccctgtgtg ttctcgttat gttgaggaaa aaaataatgg ttgctaagag attcgaactc
     5521 ttgcatctta cgatacctga gtattcccac agttaactgc ggtcaagata tttcttgaat
     5581 caggcgcctt agaccgctcg gccaaacaac caattacttg ttgagaaata gagtataatt
     5641 atcctataaa tataacgttt ttgaacacac atgaacaagg aagtacagga caattgattt
     5701 tgaagagaat gtggattttg atgtaattgt tgggattcca tttttaataa ggcaataata
     5761 ttaggtatgt ggatatacta gaagttctcc tcgaccgtcg atatgcggtg tgaaataccg
     5821 cacagatgcg taaggagaaa ataccgcatc aggaaattgt aaacgttaat attttgttaa
     5881 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     5941 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     6001 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     6061 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     6121 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     6181 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     6241 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     6301 agggcgcgtc gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     6361 gggcctcttc gctattacgc cagctggcga aggggggatg tgctgcaagg cgattaagtt
     6421 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattg
//
