LOCUS       sequence-71256-e        4444 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides a D-alanine resistance cassette as GreenGate module..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4444)
  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 4444)
  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: pGGF003
FEATURES             Location/Qualifiers
     source          1..4444
                     /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"
     promoter        2254..2634
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     terminator      3757..4009
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     primer_bind     complement(4030..4049)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4031..4049)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4059..4081)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4275..4294)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4394..4416
                     /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 ggtctcaact agtttttcaa atcagtgcgc aagacgtgac
     2281 gtaagtatcc gagtcagttt ttatttttct actaatttgg tcgtttattt cggcgtgtag
     2341 gacatggcaa ccgggcctga atttcgcggg tattctgttt ctattccaac tttttcttga
     2401 tccgcagcca ttaacgactt ttgaatagat acgctgacac gccaagcctc gctagtcaaa
     2461 agtgtaccaa acaacgcttt acagcaagaa cggaatgcgc gtgacgctcg cggtgacgcc
     2521 atttcgcctt ttcagaaatg gataaatagc cttgcttcct attatatctt cccaaattac
     2581 caatacatta cactagcatc tgaatttcat aaccaatctc gatacaccaa atcgggtacc
     2641 aacaatgcac tcgcagaagc gcgtcgttgt cctcggatca ggcgttatcg gtctgagcag
     2701 cgccctcatc ctcgctcgga agggctacag cgtgcatatt ctcgcgcgcg acttgccgga
     2761 ggacgtgtcg agccagactt tcgcttcacc ttgggctggc gcgaattgga cgcctttcat
     2821 gacgcttaca gacggtcctc gacaagcaaa atgggaagaa tcgactttca agaagtgggt
     2881 cgagttggtc ccgacgggcc atgccatgtg gctcaagggg acgaggcggt tcgcgcagaa
     2941 cgaagacggc ttgctcgggc actggtacaa ggacatcacg ccaaattacc gccccctccc
     3001 atcttccgaa tgtccacctg gcgctatcgg cgtaacctac gacaccctct ccgtccacgc
     3061 accaaagtac tgccagtacc ttgcaagaga gctacagaag ctcggcgcga cgtttgagag
     3121 acggaccgtt acgtcgcttg agcaggcgtt cgacggtgcg gatttggtgg tcaacgctac
     3181 gggacttggc gccaagtcga ttgcgggcat cgacgaccaa gccgccgagc caatccgcgg
     3241 gcaaaccgtc ctcgtcaagt ccccatgcaa gcgatgcacg atggactcgt ccgaccccgc
     3301 ttctcccgcc tacatcattc cccgaccagg tggcgaagtc atctgcggcg ggacgtacgg
     3361 cgtgggagac tgggacttgt ctgtcaaccc agagacggtc cagcgcatcc tcaagcactg
     3421 cttgcgcctc gacccgacca tctcgtcgga cggaacgatc gaaggcatcg aggtcctccg
     3481 ccacaacgtc ggcttgcgac ctgcacgacg aggcggaccc cgcgttgagg cagaacggat
     3541 cgtcctgcct ctcgaccgga caaagtcgcc cctctcgctc ggcaggggca gcgcacgagc
     3601 ggcgaaggag aaggaggtca cgcttgtgca tgcgtatggc ttctccagtg cgggatacca
     3661 gcagagttgg ggcgcggcgg aggatgtcgc gcagctcgtg gacgaggcgt tccagcgcta
     3721 ccacggcgcg gcgcgggagt cgaagttgta ggagctcttg gactcccatg ttggcaaagg
     3781 caaccaaaca aacaatgaat gatccgctcc tgcatatggg gcggtttgag tatttcaact
     3841 gccatttggg ctgaattgaa gacatgctcc tgtcagaaat tccgtgatct tactcaatat
     3901 tcagtaatct cggccaatat cctaaatgtg cgtggcttta tctgtctttg tattgtttca
     3961 tcaattcatg taacgtttgc ttttcttatg aattttcaaa taaattatcg tattgagacc
     4021 gaattctcgc cctatagtga gtcgtattac ctggccgtcg ttttacaacg tcgtgactgg
     4081 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     4141 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     4201 gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     4261 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     4321 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     4381 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     4441 gcga
//
