LOCUS       sequence-71228-e        4953 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides 3xGFP as GreenGate coding sequence module..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4953)
  AUTHORS   Lampropoulos A, Sutikovic Z, Wenzl C, Maegele I, Lohmann JU, Forner 
            J
  TITLE     GreenGate - A Novel, Versatile, and Efficient Cloning System for 
            Plant Transgenesis.
  JOURNAL   PLoS One. 2013 Dec 20;8(12):e83043. doi: 
            10.1371/journal.pone.0083043.
  PUBMED    24376629
REFERENCE   2  (bases 1 to 4953)
  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: "PLoS One.";
            date: "2013-12-20"; volume: "8(12):e83043. doi"; pages: " 
            10.1371/journal.pone.0083043"
COMMENT     Sequence Label: pGGC025
FEATURES             Location/Qualifiers
     source          1..4953
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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        2212..2230
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2212..2229
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             join(3802..3804,3805..3807,3808..4518)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(3847..3868)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4108..4127)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4455..4476
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4539..4558)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4540..4558)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4568..4590)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4784..4803)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4903..4925
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtggctct cgaggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacgagc tatttaggtg
     2221 acactataga agtgaagctt ggtctcaggc tccatggttt ctaaaggtga agagttgttt
     2281 actggagttg tgcctattct cgttgaactt gatggagatg tgaacggaca caagttctct
     2341 gtttcaggag aaggagaggg agatgctact tatggaaaac ttactttgaa gtttatttgt
     2401 actactggaa aacttcctgt tccttggcct actttggtga ctactcttac ttatggagtg
     2461 caatgcttct caagataccc agatcacatg aagcagcacg atttcttcaa atctgctatg
     2521 cctgaaggtt atgttcaaga gagaacaatt ttctttaagg atgatggaaa ctacaaaact
     2581 agagctgagg ttaagtttga gggagataca ttggtgaata gaattgaact caagggaatc
     2641 gatttcaaag aggatggtaa catccttgga cataaattgg aatacaacta caactcacac
     2701 aacgtttaca tcatggctga taagcaaaag aacggaatta aggtgaactt caagatcaga
     2761 cataacatcg aggatggatc tgttcagttg gctgatcact atcaacagaa tactccaatt
     2821 ggagatggac cagttctttt gcctgataac cattaccttt ctacacagtc aaagctgtct
     2881 aaagatccta atgagaaaag agatcacatg gttctccttg agtttgtgac tgctgcaggt
     2941 atcacactcg gtatggatga gctttacaag ggtacctctg gatctggatc aggttctggt
     3001 agtggttctg gttcaggatc tgcatgcatg gtttctaagg gtgaggaact cttcactggt
     3061 gttgttccta tcctcgttga gcttgatggt gatgtgaacg gacataagtt ctctgtttct
     3121 ggtgaaggtg agggtgatgc tacttacgga aagcttactc tcaagttcat ctgtaccact
     3181 ggaaagttgc ctgttccttg gcctactctc gttactactt tcacttacgg tgtgcagtgc
     3241 ttctcaagat accctgatca tatgaagcag cacgatttct tcaagtctgc tatgcctgag
     3301 ggatacgtgc aagagagaac tatcttcttc aaggatgatg gaaactacaa gaccagggct
     3361 gaggttaagt tcgagggtga tactcttgtg aacaggatcg agcttaaggg aatcgatttc
     3421 aaagaggatg gtaacatcct tggacacaag cttgagtaca actacaactc tcacaacgtg
     3481 tacatcatgg ctgataagca gaagaacgga atcaaggtta acttcaagat caggcacaac
     3541 atcgaggatg gatctgttca gcttgctgat cattaccagc agaacactcc tattggagat
     3601 ggacctgttc ttctccctga taaccattac ctttctaccc agtctaagct ctctaaagat
     3661 cctaacgaga agagggatca tatggtgctc ttggagtttg ttactgctgc tggaatcact
     3721 cacggaatgg atgaacttta caagccgcgg tctggaagtg gtagtggaag tggatcaggt
     3781 agtggatctg gaagtaccgg tatggtgagc aagggcgagg agctgttcac cggggtggtg
     3841 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     3901 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     3961 ctgcccgtgc cctggcccac cctcgtgacc accttcacct acggcgtgca gtgcttcagc
     4021 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     4081 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     4141 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     4201 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     4261 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     4321 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     4381 gtgctgctgc ccgacaacca ctacctgagc acccagtcca agctgagcaa agaccccaac
     4441 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactcacggc
     4501 atggacgagc tgtacaagtc agtgagaccg aattctcgcc ctatagtgag tcgtattacc
     4561 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     4621 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     4681 cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta
     4741 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
     4801 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     4861 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     4921 agaggttttc accgtcatca ccgaaacgcg cga
//
