LOCUS       sequence-258693-        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"
     protein_bind    7..28
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(43..48,49..66,67..73)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    81..97
                     /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        111..129
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     111..128
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             153..848
                     /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(286..304)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     554..573
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(872..891)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(873..891)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(901..923)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1117..1136)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1236..1258
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1296..1314)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1382..1486
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1487..1555,1556..2347)
                     /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(1705..1724)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2518..3106
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3007..3026
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3260..3277
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
       61 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacgagct atttaggtga
      121 cactatagaa gtgaagcttg gtctcatcag caatggtgag caagggcgag gcagtgatca
      181 aggagttcat gcggttcaag gtgcacatgg agggctccat gaacggccac gagttcgaga
      241 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
      301 ccaagggtgg ccccctgccc ttctcctggg acatcctgtc ccctcagttc atgtacggct
      361 ccagggcctt caccaagcac cccgccgaca tccccgacta ctataagcag tccttccccg
      421 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgccgtg accgtgaccc
      481 aggacacctc cctggaggac ggcaccctga tctacaaggt gaagctccgc ggcaccaact
      541 tccctcctga cggccccgta atgcagaaga agacaatggg ctgggaagcg tccaccgagc
      601 ggttgtaccc cgaggacggc gtgctgaagg gcgacattaa gatggccctg cgcctgaagg
      661 acggcggccg ctacctggcg gacttcaaga ccacctacaa ggccaagaag cccgtgcaga
      721 tgcccggcgc ctacaacgtc gaccgcaagt tggacatcac ctcccacaac gaggactaca
      781 ccgtggtgga acagtacgaa cgctccgagg gccgccactc caccggcggc atggacgagc
      841 tgtacaagta actgctgaga ccgaattctc gccctatagt gagtcgtatt acctggccgt
      901 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
      961 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     1021 acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct
     1081 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
     1141 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     1201 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     1261 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     1321 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     1381 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     1441 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     1501 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     1561 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     1621 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     1681 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     1741 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     1801 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     1861 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     1921 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1981 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     2041 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     2101 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     2161 tggctggttt attgctgata aatctggagc cggtgagcgt ggctctcgag gtatcattgc
     2221 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     2281 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     2341 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     2401 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     2461 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2521 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2581 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2641 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     2701 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2761 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2821 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     2881 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2941 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3001 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3061 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3121 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     3181 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     3241 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     3301 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     3361 cgactggaaa gcgggcagtg agcgcaa
//
