LOCUS       sequence-258691-        3402 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  contains the sequence of mRuby2 for 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: pGD253
FEATURES             Location/Qualifiers
     source          1..3402
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(15..20,21..38,39..45)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    53..69
                     /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        83..101
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     83..100
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             125..835
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     primer_bind     complement(859..878)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(860..878)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(888..910)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1104..1123)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1223..1245
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1283..1301)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1369..1473
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1474..1542,1543..2334)
                     /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(1692..1711)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2505..3093
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2994..3013
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3247..3264
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3381..3402
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
       61 ggataacaat ttcacacgag ctatttaggt gacactatag aagtgaagct tggtctcatc
      121 aggaatggtg tctaagggcg aagagctgat caaggaaaat atgcgtatga aggtggtcat
      181 ggaaggttcg gtcaacggcc accaattcaa atgcacaggt gaaggagaag gcaatccgta
      241 catgggaact caaaccatga ggatcaaagt catcgaggga ggacccctgc catttgcctt
      301 tgacattctt gccacgtcgt tcatgtatgg cagccgtact tttatcaagt acccgaaagg
      361 cattcctgat ttctttaaac agtcctttcc tgagggtttt acttgggaaa gagttacgag
      421 atacgaagat ggtggagtcg tcaccgtcat gcaggacacc agccttgagg atggctgtct
      481 cgtttaccac gtccaagtca gaggggtaaa ctttccctcc aatggtcccg tgatgcagaa
      541 gaagaccaag ggttgggagc ctaatacaga gatgatgtat ccagcagatg gtggtctgag
      601 gggatacact catatggcac tgaaagttga tggtggtggc catctgtctt gctctttcgt
      661 aacaacttac aggtcaaaaa agaccgtcgg gaacatcaag atgcccggta tccatgccgt
      721 tgatcaccgc ctggaaaggt tagaggaaag tgacaatgaa atgttcgtag tacaacgcga
      781 acacgcagtt gccaagttcg ccgggcttgg tggtgggatg gacgagctgt acaagtaact
      841 gctgagaccg aattctcgcc ctatagtgag tcgtattacc tggccgtcgt tttacaacgt
      901 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
      961 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     1021 ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     1081 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     1141 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     1201 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     1261 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     1321 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     1381 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     1441 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     1501 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     1561 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     1621 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     1681 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     1741 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     1801 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     1861 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     1921 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     1981 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     2041 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     2101 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     2161 gctgataaat ctggagccgg tgagcgtggc tctcgaggta tcattgcagc actggggcca
     2221 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     2281 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     2341 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     2401 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     2461 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     2521 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     2581 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     2641 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     2701 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     2761 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     2821 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     2881 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     2941 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     3001 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     3061 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     3121 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     3181 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     3241 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     3301 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     3361 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc at
//
