LOCUS       sequence-254358-        3387 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  contains the sequence of mScarlet 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 3387)
  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 3387)
  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: pRD134
FEATURES             Location/Qualifiers
     source          1..3387
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(21..40)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(22..40)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(50..72)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(266..285)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     385..407
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(445..463)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        531..635
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(636..704,705..1496)
                     /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(854..873)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1667..2255
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2156..2175
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2409..2426
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2543..2564
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2579..2584,2585..2602,2603..2609)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2617..2633
                     /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        2647..2665
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2647..2664
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             2689..3384
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(2822..2840)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3090..3109
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 ctgctgagac cgaattctcg ccctatagtg agtcgtatta cctggccgtc gttttacaac
       61 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
      121 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
      181 gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
      241 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
      301 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
      361 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
      421 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
      481 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
      541 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
      601 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
      661 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
      721 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
      781 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
      841 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
      901 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
      961 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     1021 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     1081 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     1141 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     1201 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     1261 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     1321 ttgctgataa atctggagcc ggtgagcgtg gctctcgagg tatcattgca gcactggggc
     1381 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     1441 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     1501 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     1561 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     1621 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     1681 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     1741 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     1801 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     1861 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     1921 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     1981 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     2041 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     2101 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     2161 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     2221 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     2281 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     2341 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     2401 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     2461 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     2521 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     2581 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     2641 cgagctattt aggtgacact atagaagtga agcttggtct catcagcaat ggtgagcaag
     2701 ggcgaggcag tgatcaagga gttcatgcgg ttcaaggtgc acatggaggg ctccatgaac
     2761 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     2821 gccaagctga aggtgaccaa gggtggcccc ctgcccttct cctgggacat cctgtcccct
     2881 cagttcatgt acggctccag ggccttcacc aagcaccccg ccgacatccc cgactactat
     2941 aagcagtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
     3001 gccgtgaccg tgacccagga cacctccctg gaggacggca ccctgatcta caaggtgaag
     3061 ctccgcggca ccaacttccc tcctgacggc cccgtaatgc agaagaagac aatgggctgg
     3121 gaagcgtcca ccgagcggtt gtaccccgag gacggcgtgc tgaagggcga cattaagatg
     3181 gccctgcgcc tgaaggacgg cggccgctac ctggcggact tcaagaccac ctacaaggcc
     3241 aagaagcccg tgcagatgcc cggcgcctac aacgtcgacc gcaagttgga catcacctcc
     3301 cacaacgagg actacaccgt ggtggaacag tacgaacgct ccgagggccg ccactccacc
     3361 ggcggcatgg acgagctgta caagtaa
//
