LOCUS       sequence-254347-        3435 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  contains the sequence of eGFP 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: pGD165
FEATURES             Location/Qualifiers
     source          1..3435
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(34..36,37..39,40..753)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(79..100)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(340..359)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     687..708
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(774..793)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(775..793)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(803..825)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1019..1038)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1138..1160
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1198..1216)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1284..1388
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1389..1457,1458..2249)
                     /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(1607..1626)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2420..3008
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2909..2928
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3162..3179
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3296..3317
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3332..3337,3338..3355,3356..3362)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3370..3386
                     /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        3400..3418
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3400..3417
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 tcaggcagcg gctctggatc ggcggccgct gcaatggtga gcaagggcga ggagctgttc
       61 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      121 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
      181 accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcac ctacggcgtg
      241 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
      301 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
      361 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
      421 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
      481 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
      541 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
      601 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
      661 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
      721 atcactcacg gcatggacga gctgtacaag tagctgctga gaccgaattc tcgccctata
      781 gtgagtcgta ttacctggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
      841 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
      901 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc
      961 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta
     1021 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     1081 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     1141 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     1201 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     1261 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     1321 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     1381 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     1441 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     1501 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     1561 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     1621 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     1681 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     1741 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     1801 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     1861 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     1921 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     1981 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     2041 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     2101 gtggctctcg aggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     2161 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     2221 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     2281 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     2341 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     2401 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     2461 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     2521 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     2581 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     2641 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     2701 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     2761 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     2821 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     2881 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     2941 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3001 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     3061 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     3121 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     3181 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     3241 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     3301 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     3361 tgtgtggaat tgtgagcgga taacaatttc acacgagcta tttaggtgac actatagaag
     3421 tgaagcttgg tctca
//
