LOCUS       sequence-258686-        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"
     protein_bind    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(37..42,43..60,61..67)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /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        105..123
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     105..122
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             join(174..176,177..179,180..893)
                     /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(219..240)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(480..499)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     827..848
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(914..933)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(915..933)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(943..965)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1159..1178)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1278..1300
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1338..1356)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1424..1528
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1529..1597,1598..2389)
                     /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(1747..1766)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2560..3148
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3049..3068
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3302..3319
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacacg agctatttag gtgacactat
      121 agaagtgaag cttggtctca tcaggcagcg gctctggatc ggcggccgct gcaatggtga
      181 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
      241 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
      301 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
      361 ccaccttcac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
      421 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
      481 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
      541 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
      601 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
      661 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
      721 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
      781 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
      841 agttcgtgac cgccgccggg atcactcacg gcatggacga gctgtacaag tagctgctga
      901 gaccgaattc tcgccctata gtgagtcgta ttacctggcc gtcgttttac aacgtcgtga
      961 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     1021 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     1081 tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     1141 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     1201 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     1261 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     1321 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     1381 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     1441 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     1501 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     1561 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     1621 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     1681 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     1741 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     1801 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     1861 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     1921 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     1981 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     2041 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     2101 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     2161 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     2221 taaatctgga gccggtgagc gtggctctcg aggtatcatt gcagcactgg ggccagatgg
     2281 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     2341 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     2401 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     2461 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     2521 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     2581 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     2641 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     2701 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     2761 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     2821 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     2881 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     2941 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3001 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3061 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3121 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3181 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     3241 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     3301 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     3361 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     3421 tgagcgcaac gcaat
//
