LOCUS       40924_48088        6752 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast integrative vector pYFP-Lim4-CFP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6752)
  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  JOURNAL   Unpublished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 6752)
  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 6752)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6752)
  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            "Unpublished"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..6752
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             9..719
                     /codon_start=1
                     /label=CFP
                     /note="cyan variant of GFP"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKAH
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     terminator      850..1039
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(1062..1080)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1101..1117)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1125..1141)
                     /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(1149..1179)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1194..1215)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1503..2091)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2265..3122)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3123..3227)
                     /label=AmpR promoter
     promoter        3512..3732
                     /label=URA3 promoter
     CDS             3733..4533
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(4665..5120)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     5265..5281
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5288..5306
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        5321..5717
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             5753..5773
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             5783..5803
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             5807..5827
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             5837..6553
                     /codon_start=1
                     /label=EYFP
                     /note="enhanced YFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     CDS             6596..6607
                     /codon_start=1
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
ORIGIN
        1 ctagcgccag taaaggagaa gaacttttca ctggagttgt cccaattctt gttgaattag
       61 atggtgatgt taatgggcac aaattttctg tcagtggaga gggtgaaggt gatgcaacat
      121 acggaaaact tacccttaaa tttatttgca ctactggaaa actacctgtt ccatggccaa
      181 cacttgtcac tactttgact tggggtgttc aatgcttttc aagataccca gatcatatga
      241 aacggcatga ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa agaactatat
      301 ttttcaaaga tgacgggaac tacaagacac gtgctgaagt caagtttgaa ggtgataccc
      361 ttgttaatag aatcgagtta aaaggtattg attttaaaga agatggaaac attcttggac
      421 acaaattgga atacaactat atttcacaca atgtatacat cactgcagac aaacaaaaga
      481 atggaatcaa agctcatttc aaaattagac acaacattga agatggaagc gttcaactag
      541 cagaccatta tcaacaaaat actccaattg gcgatggccc tgtcctttta ccagacaacc
      601 attacctgtc cacacaatct gccctttcga aagatcccaa cgaaaagaga gaccacatgg
      661 tccttcttga gtttgtaaca gctgctggga ttacacatgg catggatgaa ctatacaaat
      721 agggcgcgcc acttctaaat aagcgaattt cttatgattt ataagggcga attccagcac
      781 actggcggcc gttactagtc gagtcatgta attagttatg tcacgcttac attcacgccc
      841 tccccccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc
      901 tatttatttt tttatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct
      961 tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct tgcttgagaa
     1021 ggttttggga cgctcgaagg cttggtaccc agcttttgtt ccctttagtg agggttaatt
     1081 ccgagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     1141 attccacaca acataggagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     1201 aggtaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     1261 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     1321 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     1381 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     1441 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     1501 tttttccata ggctcggccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     1561 tggcgaaacc cgacaggact ataaagatac caggcgttcc cccctggaag ctccctcgtg
     1621 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     1681 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     1741 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     1801 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     1861 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     1921 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     1981 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     2041 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2101 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     2161 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     2221 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     2281 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actgcccgtc
     2341 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     2401 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     2461 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     2521 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     2581 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     2641 tcaaggcgag ttacatgatc ccccatgttg tgaaaaaaag cggttagctc cttcggtcct
     2701 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     2761 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     2821 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     2881 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     2941 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     3001 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     3061 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     3121 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     3181 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     3241 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     3301 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     3361 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     3421 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     3481 gagcagattg tactgagagt gcaccacgct tttcaattca attcatcatt ttttttttat
     3541 tctttttttt gatttcggtt tctttgaaat ttttttgatt cggtaatctc cgaacagaag
     3601 gaagaacgaa ggaaggagca cagacttaga ttggtatata tacgcatatg tagtgttgaa
     3661 gaaacatgaa attgcccagt attcttaacc caactgcaca gaacaaaaac ctgcaggaaa
     3721 cgaagataaa tcatgtcgaa agctacatat aaggaacgtg ctgctactca tcctagtcct
     3781 gttgctgcca agctatttaa tatcatgcac gaaaagcaaa caaacttgtg tgcttcattg
     3841 gatgttcgta ccaccaagga attactggag ttagttgaag cattaggtcc caaaatttgt
     3901 ttactaaaaa cacatgtgga tatcttgact gatttttcca tggagggcac agttaagccg
     3961 ctaaaggcat tatccgccaa gtacaatttt ttactcttcg aagacagaaa atttgctgac
     4021 attggtaata cagtcaaatt gcagtactct gcgggtgtat acagaatagc agaatgggca
     4081 gacattacga atgcacacgg tgtggtgggc ccaggtattg ttagcggttt gaagcaggcg
     4141 gcagaagaag taacaaagga acctagaggc cttttgatgt tagcagaatt gtcatgcaag
     4201 ggctccctat ctactggaga atatactaag ggtactgttg acattgcgaa gagcgacaaa
     4261 gattttgtta tcggctttat tgctcaaaga gacatgggtg gaagagatga aggttacgat
     4321 tggttgatta tgacacccgg tgtgggttta gatgacaagg gagacgcatt gggtcaacag
     4381 tatagaaccg tggatgatgt ggtctctaca ggatctgaca ttattattgt tggaagagga
     4441 ctatttgcaa agggaaggga tgctaaggta gagggtgaac gttacagaaa agcaggctgg
     4501 gaagcatatt tgagaagatg cggccagcaa aactaaaaaa ctgtattata agtaaatgca
     4561 tgtatactaa actcacaaat tagagcttca atttaattat atcagttatt accctgcggt
     4621 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggaaattg taaacgttaa
     4681 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     4741 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     4801 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     4861 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     4921 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     4981 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     5041 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     5101 tgcgccgcta cagggcgcgt cgcgccattc gccattcagg ctgcgcaact gttgggaagg
     5161 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaggggggat gtgctgcaag
     5221 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     5281 tgaattgtaa tacgactcac tatagggcga attggagctc cagcttcaaa atgtttctac
     5341 tcctttttta ctcttccaga ttttctcgga ctccgcgcat cgccgtacca cttcaaaaca
     5401 cccaagcaca gcatactaaa tttcccctct ttcttcctct agggtgtcgt taattacccg
     5461 tactaaaggt ttggaaaaga aaaaagagac cgcctcgttt ctttttcttc gtcgaaaaag
     5521 gcaataaaaa tttttatcac gtttcttttt cttgaaaatt ttttttttga tttttttctc
     5581 tttcgatgac ctcccattga tatttaagtt aataaacggt cttcaatttc tcaagtttca
     5641 gtttcatttt tcttgttcta ttacaacttt ttttacttct tgctcattag aaagaaagca
     5701 tagcaatcta atctaagttt tctagatatc tgcagaattc gcccttatgg ttccaaagaa
     5761 gaagagaaag gttttaattg atccaaaaaa gaaaagaaag gttgatccaa agaaaaagag
     5821 aaaagtttta attaacatgg tgagtaaagg agaagaactt ttcactggag ttgtcccaat
     5881 tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga
     5941 aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc
     6001 tgttccatgg ccaacacttg tcactacttt cggttatggt ctaaaatgct ttgctagata
     6061 cccagatcat atgaaacggc atgacttttt caagagtgcc atgcccgaag gttatgtaca
     6121 ggaaagaact atatttttca aagatgacgg gaactacaag acacgtgctg aagtcaagtt
     6181 tgaaggtgat acccttgtta atagaatcga gttaaaaggt attgatttta aagaagatgg
     6241 aaacattctt ggacacaaat tggaatacaa ctataactca cacaatgtat acatcatggc
     6301 agacaaacaa aagaatggaa tcaaagttaa cttcaaaatt agacacaaca ttgaagatgg
     6361 aagcgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct
     6421 tttaccagac aaccattacc tgtcctatca atctgccctt tcgaaagatc ccaacgaaaa
     6481 gagagaccac atggtccttc ttgagtttgt aacagctgct gggattacac atggcatgga
     6541 tgaactatac aaaggcggat ccggggtccc catctgtggt gcttgccgac ggcccatcga
     6601 agggcgcgtg gtgaacgcta tgggcaagca gtggcatgtg gagcattttg tttgtgccaa
     6661 gtgtgagaaa ccctttcttg gacatcgcca ttatgagagg aaaggcctgg catattgtga
     6721 aactcactat aaccagctat ttggtgatgt tg
//
