LOCUS       sequence-258700-        3402 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  contains the sequence of mRuby3for C-term fusions in the pGGD000 
            backbone (GreenGate).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3402)
  AUTHORS   Denay G, Schultz P, Hansch S, Weidtkamp-Peters S, Simon R
  TITLE     Over the rainbow: A practical guide for fluorescent protein 
            selection in plant FRET experiments.
  JOURNAL   Plant Direct. 2019 Dec 6;3(12):e00189. doi: 10.1002/pld3.189. 
            eCollection 2019 Dec.
  PUBMED    31844834
REFERENCE   2  (bases 1 to 3402)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant 
            Direct."; date: "2019-12-6"; volume: "3(12):e00189. doi"; pages: " 
            10.1002/pld3.189. eCollection 2019 Dec"
COMMENT     Sequence Label: pGD431
FEATURES             Location/Qualifiers
     source          1..3402
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(37..42,43..60,61..67)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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        105..123
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     105..122
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             147..857
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTD
                     IALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAV
                     AKYSNLGGGMDELYK"
     primer_bind     complement(881..900)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(882..900)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(910..932)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1126..1145)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1245..1267
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1305..1323)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1391..1495
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1496..1564,1565..2356)
                     /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     complement(1714..1733)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2527..3115
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3016..3035
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3269..3286
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacacg agctatttag gtgacactat
      121 agaagtgaag cttggtctca tcaggaatgg tgtctaaggg cgaagagctg atcaaggaaa
      181 atatgcgtat gaaggtggtc atggaaggtt cggtcaacgg ccaccaattc aaatgcacag
      241 gtgaaggaga aggcagaccg tacgagggaa ctcaaaccat gaggatcaaa gtcatcgagg
      301 gaggacccct gccatttgcc tttgacattc ttgccacgtc gttcatgtat ggcagccgta
      361 cttttatcaa gtacccggcc gacatccctg atttctttaa acagtccttt cctgagggtt
      421 ttacttggga aagagttacg agatacgaag atggtggagt cgtcaccgtc acgcaggaca
      481 ccagccttga ggatggcgag ctcgtctaca acgtcaaggt cagaggggta aactttccct
      541 ccaatggtcc cgtgatgcag aagaagacca agggttggga gcctaataca gagatgatgt
      601 atccagcaga tggtggtctg agaggataca ctgacatcgc actgaaagtt gatggtggtg
      661 gccatctgca ctgcaacttc gtgacaactt acaggtcaaa aaagaccgtc gggaacatca
      721 agatgcccgg tgtccatgcc gttgatcacc gcctggaaag gatcgaggag agtgacaatg
      781 aaacctacgt agtgcaacgc gaagtggcag ttgccaaata cagcaacctt ggtggtggca
      841 tggacgagct gtacaagtaa ctgctgagac cgaattctcg ccctatagtg agtcgtatta
      901 cctggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
      961 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     1021 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct
     1081 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
     1141 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     1201 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     1261 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     1321 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     1381 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     1441 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     1501 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     1561 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     1621 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     1681 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     1741 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     1801 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     1861 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     1921 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     1981 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     2041 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     2101 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     2161 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg gctctcgagg
     2221 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     2281 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     2341 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     2401 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     2461 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2521 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2581 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2641 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     2701 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2761 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2821 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2881 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     2941 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3001 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3061 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3121 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     3181 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     3241 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     3301 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     3361 caggtttccc gactggaaag cgggcagtga gcgcaacgca at
//
