LOCUS       sequence-127202-        4870 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides the CDS of the beta-glucuronidase gene as a level 0 
            GoldenBraid part.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4870)
  AUTHORS   Sarrion-Perdigones A, Vazquez-Vilar M, Palaci J, Castelijns B, 
            Forment J, Ziarsolo P, Blanca J, Granell A, Orzaez D
  TITLE     GoldenBraid 2.0: a comprehensive DNA assembly framework for plant 
            synthetic biology.
  JOURNAL   Plant Physiol. 2013 Jul;162(3):1618-31.
  PUBMED    23669743
REFERENCE   2  (bases 1 to 4870)
  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 
            Physiol."; date: "2013-07"; volume: "162(3)"; pages: "1618-31"
COMMENT     Sequence Label: pGUS (GB0208)
FEATURES             Location/Qualifiers
     source          1..4870
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             6..1817
                     /codon_start=1
                     /gene="uidA (or gusA) from E. coli"
                     /product="beta-glucuronidase"
                     /label=GUS
                     /translation="MLRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     promoter        complement(1920..1938)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1921..1938)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1956..1972)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1956..1972)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1969..1991)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1980..1996
                     /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        complement(join(2004..2010,2011..2028,2029..2034))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2049..2070
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2187..2204)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2358..2946)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2438..2457)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3117..3908,3909..3977))
                     /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     3740..3759
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3978..4082)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4160..4615)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4297..4318)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4509..4528
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4741..4763
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4755..4772
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4756..4772
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4779..4798
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4779..4797
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4828..4837
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      4832..4841
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ctcgaatgtt acgtcctgta gaaaccccaa cccgtgaaat caaaaaactc gacggcctgt
       61 gggcattcag tctggatcgc gaaaactgtg gaattgatca gcgttggtgg gaaagcgcgt
      121 tacaagaaag ccgggcaatt gctgtgccag gcagttttaa cgatcagttc gccgatgcag
      181 atattcgtaa ttatgcgggc aacgtctggt atcagcgcga agtctttata ccgaaaggtt
      241 gggcaggcca gcgtatcgtg ctgcgtttcg atgcggtcac tcattacggc aaagtgtggg
      301 tcaataatca ggaagtgatg gagcatcagg gcggctatac gccatttgaa gccgatgtca
      361 cgccgtatgt tattgccggg aaaagtgtac gtatcaccgt ttgtgtgaac aacgaactga
      421 actggcagac tatcccgccg ggaatggtga ttaccgacga aaacggcaag aaaaagcagt
      481 cttacttcca tgatttcttt aactatgccg gaatccaccg cagcgtaatg ctctacacca
      541 cgccgaacac ctgggtggac gatatcaccg tggtgacgca tgtcgcgcaa gactgtaacc
      601 acgcgtctgt tgactggcag gtggtggcca atggtgatgt cagcgttgaa ctgcgtgatg
      661 cggatcaaca ggtggttgca actggacaag gcactagcgg gactttgcaa gtggtgaatc
      721 cgcacctctg gcaaccgggt gaaggttatc tctatgaact gtgcgtcaca gccaaaagcc
      781 agacagagtg tgatatctac ccgcttcgcg tcggcatccg gtcagtggca gtgaagggcg
      841 aacagttcct gattaaccac aaaccgttct actttactgg ctttggtcgt catgaagatg
      901 cggacttgcg tggcaaagga ttcgataacg tgctgatggt gcacgaccac gcattaatgg
      961 actggattgg ggccaactcc taccgtacct cgcattaccc ttacgctgaa gagatgctcg
     1021 actgggcaga tgaacatggc atcgtggtga ttgatgaaac tgctgctgtc ggctttaacc
     1081 tctctttagg cattggtttc gaagcgggca acaagccgaa agaactgtac agcgaagagg
     1141 cagtcaacgg ggaaactcag caagcgcact tacaggcgat taaagagctg atagcgcgtg
     1201 acaaaaacca cccaagcgtg gtgatgtgga gtattgccaa cgaaccggat acccgtccgc
     1261 aaggtgcacg ggaatatttc gcgccactgg cggaagcaac gcgtaaactc gacccgacgc
     1321 gtccgatcac ctgcgtcaat gtaatgttct gcgacgctca caccgatacc atcagcgatc
     1381 tctttgatgt gctgtgcctg aaccgttatt acggatggta tgtccaaagc ggcgatttgg
     1441 aaacggcaga gaaggtactg gaaaaagaac ttctggcctg gcaggagaaa ctgcatcagc
     1501 cgattatcat caccgaatac ggcgtggata cgttagccgg gctgcactca atgtacaccg
     1561 acatgtggag tgaagagtat cagtgtgcat ggctggatat gtatcaccgc gtctttgatc
     1621 gcgtcagcgc cgtcgtcggt gaacaggtat ggaatttcgc cgattttgcg acctcgcaag
     1681 gcatattgcg cgttggcggt aacaagaaag ggatcttcac tcgcgaccgc aaaccgaagt
     1741 cggcggcttt tctgctgcaa aaacgctgga ctggcatgaa cttcggtgaa aaaccgcagc
     1801 agggaggcaa acaatgagct tcgagaccac tcatcgccca atcactagtg aattcgcggc
     1861 cgcctgcagg tcgaccatat gggagagctc ccaacgcgtt ggatgcatag cttgagtatt
     1921 ctatagtgtc acctaaatag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     1981 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     2041 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     2101 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     2161 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     2221 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     2281 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2341 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2401 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2461 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2521 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2581 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2641 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2701 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2761 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2821 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2881 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2941 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3001 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3061 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     3121 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     3181 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3241 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3301 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     3361 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     3421 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     3481 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     3541 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     3601 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3661 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3721 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     3781 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     3841 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     3901 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     3961 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     4021 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     4081 cgcacatttc cccgaaaagt gccacctgat gcggtgtgaa ataccgcaca gatgcgtaag
     4141 gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     4201 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     4261 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     4321 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     4381 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     4441 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     4501 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     4561 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt
     4621 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     4681 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     4741 cccagtcacg acgttgtaaa acgacggcca gtgaattgta atacgactca ctatagggcg
     4801 aattgggccc gacgtcgcat gctcccggcc gccatggcgg ccgcgggaat tcgattgggc
     4861 gatgagtggt
//
