LOCUS       sequence-71232-e        4500 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides GUS as GreenGate coding sequence module..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4500)
  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 4500)
  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: pGGC051
FEATURES             Location/Qualifiers
     source          1..4500
                     /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             2257..4065
                     /codon_start=1
                     /gene="uidA (or gusA) from E. coli"
                     /product="beta-glucuronidase"
                     /label=GUS
                     /translation="MVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     primer_bind     complement(4086..4105)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4087..4105)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4115..4137)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4331..4350)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4450..4472
                     /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 tcaacaatgg tccgtcctgt agaaacccca
     2281 acccgtgaaa tcaaaaaact cgacggcctg tgggcattca gtctggatcg cgaaaactgt
     2341 ggaattgatc agcgttggtg ggaaagcgcg ttacaagaaa gccgggcaat tgctgtgcca
     2401 ggcagtttta acgatcagtt cgccgatgca gatattcgta attatgcggg caacgtctgg
     2461 tatcagcgcg aagtctttat accgaaaggt tgggcaggcc agcgtatcgt gctgcgtttc
     2521 gatgcggtca ctcattacgg caaagtgtgg gtcaataatc aggaagtgat ggagcatcag
     2581 ggcggctata cgccatttga agccgatgtc acgccgtatg ttattgccgg gaaaagtgta
     2641 cgtatcaccg tttgtgtgaa caacgaactg aactggcaga ctatcccgcc gggaatggtg
     2701 attaccgacg aaaacggcaa gaaaaagcag tcttacttcc atgatttctt taactatgcc
     2761 ggaatccatc gcagcgtaat gctctacacc acgccgaaca cctgggtgga cgatatcacc
     2821 gtggtgacgc atgtcgcgca agactgtaac cacgcgtctg ttgactggca ggtggtggcc
     2881 aatggtgatg tcagcgttga actgcgtgat gcggatcaac aggtggttgc aactggacaa
     2941 ggcactagcg ggactttgca agtggtgaat ccgcacctct ggcaaccggg tgaaggttat
     3001 ctctatgaac tgtgcgtcac agccaaaagc cagacagagt gtgatatcta cccgcttcgc
     3061 gtcggcatcc ggtcagtggc agtgaagggc gaacagttcc tgattaacca caaaccgttc
     3121 tactttactg gctttggtcg tcatgaagat gcggacttgc gtggcaaagg attcgataac
     3181 gtgctgatgg tgcacgacca cgcattaatg gactggattg gggccaactc ctaccgtacc
     3241 tcgcattacc cttacgctga agagatgctc gactgggcag atgaacatgg catcgtggtg
     3301 attgatgaaa ctgctgctgt cggctttaac ctctctttag gcattggttt cgaagcgggc
     3361 aacaagccga aagaactgta cagcgaagag gcagtcaacg gggaaactca gcaagcgcac
     3421 ttacaggcga ttaaagagct gatagcgcgt gacaaaaacc acccaagcgt ggtgatgtgg
     3481 agtattgcca acgaaccgga tacccgtccg caaggtgcac gggaatattt cgcgccactg
     3541 gcggaagcaa cgcgtaaact cgacccgacg cgtccgatca cctgcgtcaa tgtaatgttc
     3601 tgcgacgctc acaccgatac catcagcgat ctctttgatg tgctgtgcct gaaccgttat
     3661 tacggatggt atgtccaaag cggcgatttg gaaacggcag agaaggtact ggaaaaagaa
     3721 cttctggcct ggcaggagaa actgcatcag ccgattatca tcaccgaata cggcgtggat
     3781 acgttagccg ggctgcactc aatgtacacc gacatgtgga gtgaagagta tcagtgtgca
     3841 tggctggata tgtatcaccg cgtctttgat cgcgtcagcg ccgtcgtcgg tgaacaggta
     3901 tggaatttcg ccgattttgc gacctcgcaa ggcatattgc gcgttggcgg taacaagaaa
     3961 gggatcttca ctcgcgaccg caaaccgaag tcggcggctt ttctgctgca aaaacgctgg
     4021 actggcatga acttcggtga aaaaccgcag cagggaggca aacaatcagt gagaccgaat
     4081 tctcgcccta tagtgagtcg tattacctgg ccgtcgtttt acaacgtcgt gactgggaaa
     4141 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     4201 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     4261 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt
     4321 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     4381 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     4441 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
//
