LOCUS       40924_48098        6890 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast integrative vector pYFP-THP12-CFP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6890)
  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  JOURNAL   Unpublished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 6890)
  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 6890)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6890)
  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            "Unpublished"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..6890
                     /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"
     terminator      2453..2642
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(2665..2683)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2704..2720)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2728..2744)
                     /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(2752..2782)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2797..2818)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3106..3694)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3868..4725)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4726..4830)
                     /label=AmpR promoter
     promoter        5115..5335
                     /label=URA3 promoter
     CDS             5336..6136
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(6268..6723)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     6868..6884
                     /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 gatccatgga aacacctcgt gagaagctga agcagcacag cgacgcctgc
     1321 aaggctgagt ctggagtatc tgaagagtcc ctcaacaagg ttcgcaaccg tgaagaagtg
     1381 gacgacccca aactgaaaga gcatgctttc tgcatcttga agagagccgg attcatcgac
     1441 gccagtggtg aattccaact ggaccacatt aagacgaaat tcaaggaaaa ctctgagcat
     1501 ccggagaagg tcgatgactt ggttgccaaa tgtgccgtca agaaggacac tcctcagcac
     1561 tcctcagccg acttctttaa atgcgttcac gataacaggt cggctagcgc cagtaaagga
     1621 gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg
     1681 cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt
     1741 aaatttattt gcactactgg aaaactacct gttccatggc caacacttgt cactactttg
     1801 acttggggtg ttcaatgctt ttcaagatac ccagatcata tgaaacggca tgactttttc
     1861 aagagtgcca tgcccgaagg ttatgtacag gaaagaacta tatttttcaa agatgacggg
     1921 aactacaaga cacgtgctga agtcaagttt gaaggtgata cccttgttaa tagaatcgag
     1981 ttaaaaggta ttgattttaa agaagatgga aacattcttg gacacaaatt ggaatacaac
     2041 tatatttcac acaatgtata catcactgca gacaaacaaa agaatggaat caaagctcat
     2101 ttcaaaatta gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa
     2161 aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa
     2221 tctgcccttt cgaaagatcc caacgaaaag agagaccaca tggtccttct tgagtttgta
     2281 acagctgctg ggattacaca tggcatggat gaactataca aatagggcgc gccacttcta
     2341 aataagcgaa tttcttatga tttataaggg cgaattccag cacactggcg gccgttacta
     2401 gtcgagtcat gtaattagtt atgtcacgct tacattcacg ccctcccccc acatccgctc
     2461 taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat ttttttatag
     2521 ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacagac
     2581 gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga gaaggttttg ggacgctcga
     2641 aggcttggta cccagctttt gttcccttta gtgagggtta attccgagct tggcgtaatc
     2701 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatagg
     2761 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgaggtaac tcacattaat
     2821 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     2881 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     2941 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3001 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3061 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctcgg
     3121 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3181 actataaaga taccaggcgt tcccccctgg aagctccctc gtgcgctctc ctgttccgac
     3241 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3301 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3361 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3421 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3481 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3541 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3601 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3661 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3721 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3781 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3841 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3901 gatctgtcta tttcgttcat ccatagttgc ctgactgccc gtcgtgtaga taactacgat
     3961 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4021 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4081 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4141 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4201 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4261 atcccccatg ttgtgaaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4321 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4381 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4441 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4501 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4561 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4621 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4681 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4741 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4801 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     4861 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     4921 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     4981 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     5041 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     5101 agtgcaccac gcttttcaat tcaattcatc attttttttt tattcttttt tttgatttcg
     5161 gtttctttga aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga
     5221 gcacagactt agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc
     5281 agtattctta acccaactgc acagaacaaa aacctgcagg aaacgaagat aaatcatgtc
     5341 gaaagctaca tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt
     5401 taatatcatg cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa
     5461 ggaattactg gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt
     5521 ggatatcttg actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc
     5581 caagtacaat tttttactct tcgaagacag aaaatttgct gacattggta atacagtcaa
     5641 attgcagtac tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca
     5701 cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa
     5761 ggaacctaga ggccttttga tgttagcaga attgtcatgc aagggctccc tatctactgg
     5821 agaatatact aagggtactg ttgacattgc gaagagcgac aaagattttg ttatcggctt
     5881 tattgctcaa agagacatgg gtggaagaga tgaaggttac gattggttga ttatgacacc
     5941 cggtgtgggt ttagatgaca agggagacgc attgggtcaa cagtatagaa ccgtggatga
     6001 tgtggtctct acaggatctg acattattat tgttggaaga ggactatttg caaagggaag
     6061 ggatgctaag gtagagggtg aacgttacag aaaagcaggc tgggaagcat atttgagaag
     6121 atgcggccag caaaactaaa aaactgtatt ataagtaaat gcatgtatac taaactcaca
     6181 aattagagct tcaatttaat tatatcagtt attaccctgc ggtgtgaaat accgcacaga
     6241 tgcgtaagga gaaaataccg catcaggaaa ttgtaaacgt taatattttg ttaaaattcg
     6301 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     6361 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     6421 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     6481 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     6541 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     6601 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     6661 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     6721 cgtcgcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     6781 cttcgctatt acgccagctg gcgaaggggg gatgtgctgc aaggcgatta agttgggtaa
     6841 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattg
//
