LOCUS       sequence-71258-e        4869 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 4869)
  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 4869)
  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: pGGF004
FEATURES             Location/Qualifiers
     source          1..4869
                     /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        2747..3092
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     3035..3055
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     complement(4455..4474)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4456..4474)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4484..4506)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4700..4719)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4819..4841
                     /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 agtgtcccca gattagcctt ttcaatttca
     2281 gaaagaatgc taacccacag atggttagag aggcttacgc agcacgtctc atcaagacga
     2341 tctacccgag caataatctc caggaaatca aataccttcc caagaaggtt aaagatgcag
     2401 tcaaaagatt caggactaac tgcatcaaga acacagagaa agatatattt ctcaagatca
     2461 gaagtactat tccagtatgg acgattcaag gcttgcttca caaaccaagg caagtaatag
     2521 agattggagt ctctaaaaag gtagttccca ctgaatcaaa ggccatggag tcaaagattc
     2581 aaatagagga cctaacagaa ctcgccgtaa agactggcga acagttcata cagagtctct
     2641 tacgactcaa tgacaagaag aaaatcttcg tcaacatggt ggagcacgac acacttgtct
     2701 actccaaaaa tatcaaagat acagtctcag aagaccaaag ggcaattgag acttttcaac
     2761 aaagggtaat atccggaaac ctcctcggat tccattgccc agctatctgt cactttattg
     2821 tgaagatagt ggaaaaggaa ggtggctcct acaaatgcca tcattgcgat aaaggaaagg
     2881 ccatcgttga agatgcctct gccgacagtg gtcccaaaga tggaccccca cccacgagga
     2941 gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa gcaagtggat tgatgtgata
     3001 tctccactga cgtaagggat gacgcacaat cccactatcc ttcgcaagac ccttcctcta
     3061 tataaggaag ttcatttcat ttggagagaa cacgggggac ggtaccaaca atgcactcgc
     3121 agaagcgcgt cgttgtcctc ggatcaggcg ttatcggtct gagcagcgcc ctcatcctcg
     3181 ctcggaaggg ctacagcgtg catattctcg cgcgcgactt gccggaggac gtgtcgagcc
     3241 agactttcgc ttcaccttgg gctggcgcga attggacgcc tttcatgacg cttacagacg
     3301 gtcctcgaca agcaaaatgg gaagaatcga ctttcaagaa gtgggtcgag ttggtcccga
     3361 cgggccatgc catgtggctc aaggggacga ggcggttcgc gcagaacgaa gacggcttgc
     3421 tcgggcactg gtacaaggac atcacgccaa attaccgccc cctcccatct tccgaatgtc
     3481 cacctggcgc tatcggcgta acctacgaca ccctctccgt ccacgcacca aagtactgcc
     3541 agtaccttgc aagagagcta cagaagctcg gcgcgacgtt tgagagacgg accgttacgt
     3601 cgcttgagca ggcgttcgac ggtgcggatt tggtggtcaa cgctacggga cttggcgcca
     3661 agtcgattgc gggcatcgac gaccaagccg ccgagccaat ccgcgggcaa accgtcctcg
     3721 tcaagtcccc atgcaagcga tgcacgatgg actcgtccga ccccgcttct cccgcctaca
     3781 tcattccccg accaggtggc gaagtcatct gcggcgggac gtacggcgtg ggagactggg
     3841 acttgtctgt caacccagag acggtccagc gcatcctcaa gcactgcttg cgcctcgacc
     3901 cgaccatctc gtcggacgga acgatcgaag gcatcgaggt cctccgccac aacgtcggct
     3961 tgcgacctgc acgacgaggc ggaccccgcg ttgaggcaga acggatcgtc ctgcctctcg
     4021 accggacaaa gtcgcccctc tcgctcggca ggggcagcgc acgagcggcg aaggagaagg
     4081 aggtcacgct tgtgcatgcg tatggcttct ccagtgcggg ataccagcag agttggggcg
     4141 cggcggagga tgtcgcgcag ctcgtggacg aggcgttcca gcgctaccac ggcgcggcgc
     4201 gggagtcgaa gttgtaggag ctcagtccgc aaaaatcacc agtctctctc tacaaatcta
     4261 tctctctcta tttttctcca gaataatgtg tgagtagttc ccagataagg gaattagggt
     4321 tcttataggg tttcgctcat gtgttgagca tataagaaac ccttagtatg tatttgtatt
     4381 tgtaaaatac ttctatcaat aaaatttcta attcctaaaa ccaaaatcca gtgagtattg
     4441 agaccgaatt ctcgccctat agtgagtcgt attacctggc cgtcgtttta caacgtcgtg
     4501 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     4561 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     4621 atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     4681 gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     4741 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     4801 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     4861 aacgcgcga
//
