LOCUS       sequence-160072-        5449 bp    DNA     circular SYN 15-APR-2025
DEFINITION  YFP-CFP positive FRET control; his selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5449)
  AUTHORS   Hailey DW, Davis TN, Muller EG
  TITLE     Fluorescence resonance energy transfer using color variants of green
            fluorescent protein.
  JOURNAL   Methods Enzymol. 2002;351:34-49.
  PUBMED    12073355
REFERENCE   2  (bases 1 to 5449)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Methods 
            Enzymol."; date: "2002"; volume: "351"; pages: "34-49"
COMMENT     Sequence Label: pDH18
FEATURES             Location/Qualifiers
     source          1..5449
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             63..779
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     CDS             63..779
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     CDS             786..1496
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKAH
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     terminator      1520..1707
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1575..1599)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1782..2982
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        1782..2125
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2126..2779
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe his5"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=S. pombe his5
                     /note="yeast auxotrophic marker; corresponds to S. 
                     cerevisiae HIS3"
                     /translation="MGRRAFVERNTNETKISVAIALDKAPLPEESNFIDELITSKHANQ
                     KGEQVIQVDTGIGFLDHMYHALAKHAGWSLRLYSRGDLIIDDHHTAEDTAIALGIAFKQ
                     AMGNFAGVKRFGHAYCPLDEALSRSVVDLSGRPYAVIDLGLKREKVGELSCEMIPHLLY
                     SFSVAAGITLHVTCLYGSNDHHRAESAFKSLAVAMRAATSLTGSSEVPSTKGVL"
     terminator      2785..2982
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3086..3105)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3087..3105)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3192..3209)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3363..3951)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3443..3462)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4122..4913,4914..4982))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4745..4764
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4983..5087)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5155..5173
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5211..5233)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5333..5352
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(5433..5449,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(5433..5449,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 acatggtgag taaaggagaa gaacttttca ctggagttgt cccaattctt gttgaattag
      121 atggtgatgt taatgggcac aaattttctg tcagtggaga gggtgaaggt gatgcaacat
      181 acggaaaact tacccttaaa tttatttgca ctactggaaa actacctgtt ccatggccaa
      241 cacttgtcac tactttcggt tatggtctaa aatgctttgc tagataccca gatcatatga
      301 aacggcatga ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa agaactatat
      361 ttttcaaaga tgacgggaac tacaagacac gtgctgaagt caagtttgaa ggtgataccc
      421 ttgttaatag aatcgagtta aaaggtattg attttaaaga agatggaaac attcttggac
      481 acaaattgga atacaactat aactcacaca atgtatacat catggcagac aaacaaaaga
      541 atggaatcaa agttaacttc aaaattagac acaacattga agatggaagc gttcaactag
      601 cagaccatta tcaacaaaat actccaattg gcgatggccc tgtcctttta ccagacaacc
      661 attacctgtc ctatcaatct gccctttcga aagatcccaa cgaaaagaga gaccacatgg
      721 tccttcttga gtttgtaaca gctgctggga ttacacatgg catggatgaa ctatacaaag
      781 gcgccagtaa aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg
      841 gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg
      901 gaaaacttac ccttaaattt atttgcacta ctggaaaact acctgttcca tggccaacac
      961 ttgtcactac tttgacttgg ggtgttcaat gcttttcaag atacccagat catatgaaac
     1021 ggcatgactt tttcaagagt gccatgcccg aaggttatgt acaggaaaga actatatttt
     1081 tcaaagatga cgggaactac aagacacgtg ctgaagtcaa gtttgaaggt gatacccttg
     1141 ttaatagaat cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt cttggacaca
     1201 aattggaata caactatatt tcacacaatg tatacatcac tgcagacaaa caaaagaatg
     1261 gaatcaaagc tcatttcaaa attagacaca acattgaaga tggaagcgtt caactagcag
     1321 accattatca acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt
     1381 acctgtccac acaatctgcc ctttcgaaag atcccaacga aaagagagac cacatggtcc
     1441 ttcttgagtt tgtaacagct gctgggatta cacatggcat ggatgaacta tacaaatagg
     1501 gcgcgccact tctaaataag cgaatttctt atgatttatg atttttatta ttaaataagt
     1561 tataaaaaaa ataagtgtat acaaatttta aagtgactct taggttttaa aacgaaaatt
     1621 cttattcttg agtaactctt tcctgtaggt caggttgctt tctcaggtat agtatgaggt
     1681 cgctcttatt gaccacacct ctaccggcag atccgctagg gataacaggg taatatagat
     1741 ctgtttagct tgcctcgtcc ccgccgggtc acccggccag cgacatggag gcccagaata
     1801 ccctccttga cagtcttgac gtgcgcagct caggggcatg atgtgactgt cgcccgtaca
     1861 tttagcccat acatccccat gtataatcat ttgcatccat acattttgat ggccgcacgg
     1921 cgcgaagcaa aaattacggc tcctcgctgc agacctgcga gcagggaaac gctcccctca
     1981 cagacgcgtt gaattgtccc cacgccgcgc ccctgtagag aaatataaaa ggttaggatt
     2041 tgccactgag gttcttcttt catatacttc cttttaaaat cttgctagga tacagttctc
     2101 acatcacatc cgaacataaa caaccatggg taggagggct tttgtagaaa gaaatacgaa
     2161 cgaaacgaaa atcagcgttg ccatcgcttt ggacaaagct cccttacctg aagagtcgaa
     2221 ttttattgat gaacttataa cttccaagca tgcaaaccaa aagggagaac aagtaatcca
     2281 agtagacacg ggaattggat tcttggatca catgtatcat gcactggcta aacatgcagg
     2341 ctggagctta cgactttact caagaggtga tttaatcatc gatgatcatc acactgcaga
     2401 agatactgct attgcacttg gtattgcatt caagcaggct atgggtaact ttgccggcgt
     2461 taaaagattt ggacatgctt attgtccact tgacgaagct ctttctagaa gcgtagttga
     2521 cttgtcggga cggccctatg ctgttatcga tttgggatta aagcgtgaaa aggttgggga
     2581 attgtcctgt gaaatgatcc ctcacttact atattccttt tcggtagcag ctggaattac
     2641 tttgcatgtt acctgcttat atggtagtaa tgaccatcat cgtgctgaaa gcgcttttaa
     2701 atctctggct gttgccatgc gcgcggctac tagtcttact ggaagttctg aagtcccaag
     2761 cacgaaggga gtgttgtaaa gagtactgac aataaaaaga ttcttgtttt caagaacttg
     2821 tcatttgtat agttttttta tattgtagtt gttctatttt aatcaaatgt tagcgtgatt
     2881 tatatttttt ttcgcctcga catcatctgc ccagatgcga agttaagtgc gcagaaagta
     2941 atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac tgctgtcgat tcgatactaa
     3001 cgccgccatc cagtttaaac gagctcgaat tcatcgatga tatcagatcc actagtggcc
     3061 tatgcggccg cggatctgcc ggtctcccta tagtgagtcg tattaatttc gataagccag
     3121 gttaacctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3181 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3241 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3301 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3361 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3421 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3481 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3541 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3601 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3661 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3721 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3781 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     3841 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3901 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3961 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4021 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4081 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4141 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4201 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4261 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4321 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4381 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     4441 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     4501 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     4561 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     4621 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     4681 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     4741 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     4801 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     4861 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     4921 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     4981 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5041 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5101 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     5161 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     5221 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     5281 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     5341 gagcagattg tactgagagt gcaccatatg gacatattgt cgttagaacg cggctacaat
     5401 taatacataa ccttatgtat catacacata cgatttaggt gacactata
//
