LOCUS       sequence-258705-        3435 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  contains the sequence of TSapphire 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 3435)
  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 3435)
  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: pGD252
FEATURES             Location/Qualifiers
     source          1..3435
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(join(44..754,755..757,758..760))
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVMVFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSIQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(86..107)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     435..454
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     694..715
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(811..829)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(812..829)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    843..859
                     /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        complement(join(867..873,874..891,892..897))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    912..933
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1050..1067)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1221..1809)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1301..1320)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1980..2771,2772..2840))
                     /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     2603..2622
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2841..2945)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3013..3031
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3069..3091)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3191..3210
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3404..3426
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 taatacgact cactataggg cgagaattcg gtctcagcag ttacttgtac agctcgtcca
       61 tgcccagagt gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct
      121 cgttggggtc tttgctcagg gcggactgga tgctcaggta gtggttgtcg ggcagcagca
      181 cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgccgccgt
      241 cctcgatgtt gtggcggatc ttgaagttgg ccttgatgcc gttcttctgc ttgtcggcca
      301 tgatatagac gttgtggctg ttgaagttgt actccagctt gtgccccagg atgttgccgt
      361 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
      421 tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga
      481 cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc
      541 gggcgaacac catcacgccg taggagaagg tggtcacgag ggtgggccag ggcacgggca
      601 gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc
      661 cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg
      721 gcaccacccc ggtgaacagc tcctcgccct tgctcaccat tgcagcggcc gccgatccag
      781 agccgctgcc tgatgagacc aagcttcact tctatagtgt cacctaaata gctcgtgtga
      841 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
      901 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
      961 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     1021 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     1081 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     1141 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     1201 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     1261 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     1321 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     1381 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     1441 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     1501 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     1561 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     1621 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     1681 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     1741 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     1801 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     1861 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     1921 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     1981 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     2041 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     2101 agtgctgcaa tgatacctcg agagccacgc tcaccggctc cagatttatc agcaataaac
     2161 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     2221 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     2281 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     2341 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     2401 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     2461 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     2521 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     2581 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     2641 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     2701 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     2761 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     2821 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     2881 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     2941 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     3001 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     3061 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     3121 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     3181 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     3241 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc
     3301 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     3361 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     3421 aaaacgacgg ccagg
//
