LOCUS       sequence-123124-        7908 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GUS overexpression in planta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7908)
  AUTHORS   Rubio-Somoza I, Zhou CM, Confraria A, Martinho C, von Born P, 
            Baena-Gonzalez E, Wang JW, Weigel D
  TITLE     Temporal control of leaf complexity by miRNA-regulated licensing of 
            protein complexes.
  JOURNAL   Curr Biol. 2014 Nov 17;24(22):2714-9. doi: 
            10.1016/j.cub.2014.09.058. Epub 2014 Nov 6.
  PUBMED    25448000
REFERENCE   2  (bases 1 to 7908)
  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"; doi: 
            "10.1016/j.cub.2014.09.058"; journalName: "Curr Biol"; date: 
            "2014-11-17- 17"; volume: "24"; issue: "22"; pages: "2714-9"
COMMENT     Sequence Label: pPM180
FEATURES             Location/Qualifiers
     source          1..7908
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    542..564
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     terminator      complement(588..840)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(860..1411)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     promoter        complement(1453..1632)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     1869..1891
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1883..1900
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1884..1900
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1910..1929
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1910..1928
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1954..1970
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2507..2852
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2795..2815
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     protein_bind    2877..2901
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     2916..2932
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2953..4764
                     /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"
     protein_bind    complement(4840..4864)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      4871..5512
                     /label=E9 terminator
                     /note="terminator and polyadenylation signal from the pea 
                     rbcS-E9 gene"
     promoter        complement(5534..5552)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5573..5589)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5573..5589)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5586..5608)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5597..5613
                     /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(5621..5627,5628..5646,5647..5652))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5667..5688
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5805..5822)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5927..5951
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(6042..6630)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6122..6141)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6801..7589)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      join(7880..7908,1..407)
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 tttttatccc cggaagcctg tggatagagg gtagttatcc acgtgaaacc gctaatgccc
       61 cgcaaagcct tgattcacgg ggctttccgg cccgctccaa aaactatcca cgtgaaatcg
      121 ctaatcaggg tacgtgaaat cgctaatcgg agtacgtgaa atcgctaata aggtcacgtg
      181 aaatcgctaa tcaaaaaggc acgtgagaac gctaatagcc ctttcagatc aacagcttgc
      241 aaacacccct cgctccggca agtagttaca gcaagtagta tgttcaatta gcttttcaat
      301 tatgaatata tatatcaatt attggtcgcc cttggcttgt ggacaatgcg ctacgcgcac
      361 cggctccgcc cgtggacaac cgcaagcggt tgcccaccgt cgagcgccag cgcctttgcc
      421 cacaacccgg cggccggccg caacagatcg ttttataaat tttttttttt gaaaaagaaa
      481 aagcccgaaa ggcggcaacc tctcgggctt ctggatttcc gatccccgga attagagatc
      541 ttggcaggat atattgtggt gtaacgttat cagcttgcat gccggtcgat ctagtaacat
      601 agatgacacc gcgcgcgata atttatccta gtttgcgcgc tatattttgt tttctatcgc
      661 gtattaaatg tataattgcg ggactctaat cataaaaacc catctcataa ataacgtcat
      721 gcattacatg ttaattatta catgcttaac gtaattcaac agaaattata tgataatcat
      781 cgcaagaccg gcaacaggat tcaatcttaa gaaactttat tgccaaatgt ttgaacgatc
      841 tgcttgactc taggggtcat cagatttcgg tgacgggcag gaccggacgg ggcggcaccg
      901 gcaggctgaa gtccagctgc cagaaaccca cgtcatgcca gttcccgtgc ttgaagccgg
      961 ccgcccgcag catgccgcgg ggggcatatc cgagcgcctc gtgcatgcgc acgctcgggt
     1021 cgttgggcag cccgatgaca gcgaccacgc tcttgaagcc ctgtgcctcc agggacttca
     1081 gcaggtgggt gtagagcgtg gagcccagtc ccgtccgctg gtggcggggg gagacgtaca
     1141 cggtggactc ggccgtccag tcgtaggcgt tgcgtgcctt ccagggaccc gcgtaggcga
     1201 tgccggcgac ctcgccgtcc acctcggcga cgagccaggg atagcgctcc cgcagacgga
     1261 cgaggtcgtc cgtccactcc tgcggttcct gcggctcggt acggaagttg accgtgcttg
     1321 tctcgatgta gtggttgacg atggtgcaga ccgccggcat gtccgcctcg gtggcacggc
     1381 ggatgtcggc cgggcgtcgt tctgggctca tggtagatcc cctcgatcga gttgagagtg
     1441 aatatgagac tctaattgga taccgagggg aatttatgga acgtcagtgg agcatttttg
     1501 acaagaaata tttgctagct gatagtgacc ttaggcgact tttgaacgcg caataatggt
     1561 ttctgacgta tgtgcttagc tcattaaact ccagaaaccc gcggctcagt ggctccttca
     1621 acgttgcggt tctgtcagtt ccaaacgtaa aacggcttgt cccgcgtcat cggcgggggt
     1681 cataacgtga ctcccttaat tctccgctca tgtatcgata acattaacgt ttacaatttc
     1741 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     1801 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     1861 agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt aatacgactc
     1921 actatagggc gaattgggta ccgggccccc cctcgaggtc gacggtatcg ataagcttga
     1981 tatcgaattc ctgcagcccg ggggatccat tcggtcccca gattagcctt ttcaatttca
     2041 gaaagaatgc taacccacag atggttagag aggcttacgc agcaggtctc atcaagacga
     2101 tctacccgag caataatctc caggaaatca aataccttcc caagaaggtt aaagatgcag
     2161 tcaaaagatt caggactaac tgcatcaaga acacagagaa agatatattt ctcaagatca
     2221 gaagtactat tccagtatgg acgattcaag gcttgcttca caaaccaagg caagtaatag
     2281 agattggagt ctctaaaaag gtagttccca ctgaatcaaa ggccatggag tcaaagattc
     2341 aaatagagga cctaacagaa ctcgccgtaa agactggcga acagttcata cagagtctct
     2401 tacgactcaa tgacaagaag aaaatcttcg tcaacatggt ggagcacgac acacttgtct
     2461 actccaaaaa tatcaaagat acagtctcag aagaccaaag ggcaattgag acttttcaac
     2521 aaagggtaat atccggaaac ctcctcggat tccattgccc agctatctgt cactttattg
     2581 tgaagatagt ggaaaaggaa ggtggctcct acaaatgcca tcattgcgat aaaggaaagg
     2641 ccatcgttga agatgcctct gccgacagtg gtcccaaaga tggaccccca cccacgagga
     2701 gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa gcaagtggat tgatgtgata
     2761 tctccactga cgtaagggat gacgcacaat cccactatcc ttcgcaagac ccttcctcta
     2821 tataaggaag ttcatttcat ttggagagaa cacgggggac gagcttctag aggatcacaa
     2881 gtttgtacaa aaaagcaggc tttcgggccc cccctcgagg tcgacggtat cgataagctt
     2941 ggatccgggc ccatggtccg tcctgtagaa accccaaccc gtgaaatcaa aaaactcgac
     3001 ggcctgtggg cattcagtct ggatcgcgaa aactgtggaa ttgatcagcg ttggtgggaa
     3061 agcgcgttac aagaaagccg ggcaattgct gtgccaggca gttttaacga tcagttcgcc
     3121 gatgcagata ttcgtaatta tgcgggcaac gtctggtatc agcgcgaagt ctttataccg
     3181 aaaggttggg caggccagcg tatcgtgctg cgtttcgatg cggtcactca ttacggcaaa
     3241 gtgtgggtca ataatcagga agtgatggag catcagggcg gctatacgcc atttgaagcc
     3301 gatgtcacgc cgtatgttat tgccgggaaa agtgtacgta tcaccgtttg tgtgaacaac
     3361 gaactgaact ggcagactat cccgccggga atggtgatta ccgacgaaaa cggcaagaaa
     3421 aagcagtctt acttccatga tttctttaac tatgccggaa tccatcgcag cgtaatgctc
     3481 tacaccacgc cgaacacctg ggtggacgat atcaccgtgg tgacgcatgt cgcgcaagac
     3541 tgtaaccacg cgtctgttga ctggcaggtg gtggccaatg gtgatgtcag cgttgaactg
     3601 cgtgatgcgg atcaacaggt ggttgcaact ggacaaggca ctagcgggac tttgcaagtg
     3661 gtgaatccgc acctctggca accgggtgaa ggttatctct atgaactgtg cgtcacagcc
     3721 aaaagccaga cagagtgtga tatctacccg cttcgcgtcg gcatccggtc agtggcagtg
     3781 aagggcgaac agttcctgat taaccacaaa ccgttctact ttactggctt tggtcgtcat
     3841 gaagatgcgg acttgcgtgg caaaggattc gataacgtgc tgatggtgca cgaccacgca
     3901 ttaatggact ggattggggc caactcctac cgtacctcgc attaccctta cgctgaagag
     3961 atgctcgact gggcagatga acatggcatc gtggtgattg atgaaactgc tgctgtcggc
     4021 tttaacctct ctttaggcat tggtttcgaa gcgggcaaca agccgaaaga actgtacagc
     4081 gaagaggcag tcaacgggga aactcagcaa gcgcacttac aggcgattaa agagctgata
     4141 gcgcgtgaca aaaaccaccc aagcgtggtg atgtggagta ttgccaacga accggatacc
     4201 cgtccgcaag gtgcacggga atatttcgcg ccactggcgg aagcaacgcg taaactcgac
     4261 ccgacgcgtc cgatcacctg cgtcaatgta atgttctgcg acgctcacac cgataccatc
     4321 agcgatctct ttgatgtgct gtgcctgaac cgttattacg gatggtatgt ccaaagcggc
     4381 gatttggaaa cggcagagaa ggtactggaa aaagaacttc tggcctggca ggagaaactg
     4441 catcagccga ttatcatcac cgaatacggc gtggatacgt tagccgggct gcactcaatg
     4501 tacaccgaca tgtggagtga agagtatcag tgtgcatggc tggatatgta tcaccgcgtc
     4561 tttgatcgcg tcagcgccgt cgtcggtgaa caggtatgga atttcgccga ttttgcgacc
     4621 tcgcaaggca tattgcgcgt tggcggtaac aagaaaggga tcttcactcg cgaccgcaaa
     4681 ccgaagtcgg cggcttttct gctgcaaaaa cgctggactg gcatgaactt cggtgaaaaa
     4741 ccgcagcagg gaggcaaaca atgaatcaac aactctcctg gcgcaccatc gtcggctaca
     4801 gcctcgggaa ttgtaccgcc gccaccgcgg tggatctaga cccagctttc ttgtacaaag
     4861 tggtgatcct agctttcgtt cgtatcatcg gtttcgacaa cgttcgtcaa gttcaatgca
     4921 tcagtttcat tgcgcacaca ccagaatcct actgagtttg agtattatgg cattgggaaa
     4981 actgtttttc ttgtaccatt tgttgtgctt gtaatttact gtgtttttta ttcggttttc
     5041 gctatcgaac tgtgaaatgg aaatggatgg agaagagtta atgaatgata tggtcctttt
     5101 gttcattctc aaattaatat tatttgtttt ttctcttatt tgttgtgtgt tgaatttgaa
     5161 attataagag atatgcaaac attttgtttt gagtaaaaat gtgtcaaatc gtggcctcta
     5221 atgaccgaag ttaatatgag gagtaaaaca cttgtagttg taccattatg cttattcact
     5281 aggcaacaaa tatattttca gacctagaaa agctgcaaat gttactgaat acaagtatgt
     5341 cctcttgtgt tttagacatt tatgaacttt cctttatgta attttccaga atccttgtca
     5401 gattctaatc attgctttat aattatagtt atactcatgg atttgtagtt gagtatgaaa
     5461 atatttttta atgcatttta tgacttgcca attgattgac aacatgcatc aatcggagct
     5521 ccagcttttg ttccctttag tgagggttaa ttccgagctt ggcgtaatca tggtcatagc
     5581 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaaghc
     5641 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     5701 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     5761 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5821 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5881 ttatccacag aatcagggga taacgcagga aagaacatga aggccttgac aggatatatt
     5941 ggcgggtaaa ctaagtcgct gtatgtgttt gtttgagatc tcatgtgagc aaaaggccag
     6001 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6061 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6121 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6181 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     6241 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6301 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6361 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6421 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6481 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaag aagagttggt
     6541 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6601 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6661 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6721 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6781 gtgtaacatt ggtctagtga ttatttgccg actaccttgg tgatctcgcc tttcacgtag
     6841 tgaacaaatt cttccaactg atctgcgcgc gaggccaagc gatcttcttg tccaagataa
     6901 gcctgcctag cttcaagtat gacgggctga tactgggccg gcaggcgctc cattgcccag
     6961 tcggcagcga catccttcgg cgcgattttg ccggttactg cgctgtacca aatgcgggac
     7021 aacgtaagca ctacatttcg ctcatcgcca gcccagtcgg gcggcgagtt ccatagcgtt
     7081 aaggtttcat ttagcgcctc aaatagatcc tgttcaggaa ccggatcaaa gagttcctcc
     7141 gccgctggac ctaccaaggc aacgctatgt tctcttgctt ttgtcagcaa gatagccaga
     7201 tcaatgtcga tcgtggctgg ctcgaagata cctgcaagaa tgtcattgcg ctgccattct
     7261 ccaaattgca gttcgcgctt agctggataa cgccacggaa tgatgtcgtc gtgcacaaca
     7321 atggtgactt ctacagcgcg gagaatctcg ctctctccag gggaagccga agtttccaaa
     7381 aggtcgttga tcaaagctcg ccgcgttgtt tcatcaagcc ttacggtcac cgtaaccagc
     7441 aaatcaatat cactgtgtgg cttcaggccg ccatccactg cggagccgta caaatgtacg
     7501 gccagcaacg tcggttcgag atggcgctcg atgacgccaa ctacctctga tagttgagtc
     7561 gatacttcgg cgatcaccgc ttccctcata acaccccttg tattactgtt tatgtaagca
     7621 gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt
     7681 tgagacacaa cgtggctttg ttgaataaat cgaacttttg ctgagttgaa ggatcagatc
     7741 acgcatcttc ccgacaacgc agaccgttcc gtggcaaagc aaaagttcaa aatcaccaac
     7801 tggtccacct acaacaaagc tctcatcaac cgtggctccc tcactttctg gctggatgat
     7861 ggggcgattc aggcgatccc catccaacag cccgccgtcg agcgggct
//
