LOCUS       sequence-134161-        5122 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry clone containing the GUS enzyme. Can be used to construct 
            transcriptional reporters. For use in plants and compatible with the
            MultiSite Gateway system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5122)
  AUTHORS   Siligato R, Wang X, Yadav SR, Lehesranta S, Ma G, Ursache R, Sevilem
            I, Zhang J, Gorte M, Prasad K, Wrzaczek M, Heidstra R, Murphy A, 
            Scheres B, Mahonen AP
  TITLE     MultiSite Gateway compatible cell type-specific gene inducible 
            system for plants.
  JOURNAL   Plant Physiol. 2015 Dec 7. pii: pp.01246.2015.
  PUBMED    26644504
REFERENCE   2  (bases 1 to 5122)
  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. 2015 Dec 7. pii: pp.01246.2015."
COMMENT     Sequence Label: p221a-GUS
FEATURES             Location/Qualifiers
     source          1..5122
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(108..126)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(131..147)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(228..246)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(229..246)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(264..280)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(277..299)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    288..304
                     /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(312..318,319..336,337..342))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    357..378
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(495..512)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(666..1254)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(746..765)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1425..2216,2217..2285))
                     /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     2048..2067
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2286..2390)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2468..2923)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2605..2626)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2817..2836
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3049..3071
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3064..3080
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3087..3105
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3136..3145
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3140..3149
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     3166..3182
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3198..3297
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3300..5111
                     /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"
     protein_bind    complement(join(5113..5122,1..90))
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt
       61 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
      121 gtattacatg gtcatagctg tttcctgaat cactagtgaa ttcgcggccg cctgcaggtc
      181 gaccatatgg gagagctccc aacgcgttgg atgcatagct tgagtattct atagtgtcac
      241 ctaaatagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
      301 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
      361 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
      421 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
      481 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
      541 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
      601 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
      661 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
      721 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
      781 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
      841 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
      901 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
      961 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     1021 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     1081 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     1141 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     1201 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     1261 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     1321 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     1381 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     1441 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     1501 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     1561 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     1621 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     1681 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     1741 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     1801 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     1861 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     1921 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     1981 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     2041 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     2101 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     2161 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     2221 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     2281 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     2341 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     2401 cgaaaagtgc cacctgatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     2461 catcaggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     2521 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     2581 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     2641 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     2701 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     2761 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     2821 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     2881 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtccattcg ccattcaggc
     2941 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     3001 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     3061 gttgtaaaac gacggccagt gaattgtaat acgactcact atagggcgaa ttgggcccga
     3121 cgtcgcatgc tcccggccgc catggcggcc gcgggaattc gatttgtaaa acgacggcca
     3181 gtcttaagct cgggccccaa ataatgattt tattttgact gatagtgacc tgttcgttgc
     3241 aacaaattga tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa gcaggctgta
     3301 tgttacgtcc tgtagaaacc ccaacccgtg aaatcaaaaa actcgacggc ctgtgggcat
     3361 tcagtctgga tcgcgaaaac tgtggaattg atcagcgttg gtgggaaagc gcgttacaag
     3421 aaagccgggc aattgctgtg ccaggcagtt ttaacgatca gttcgccgat gcagatattc
     3481 gtaattatgc gggcaacgtc tggtatcagc gcgaagtctt tataccgaaa ggttgggcag
     3541 gccagcgtat cgtgctgcgt ttcgatgcgg tcactcatta cggcaaagtg tgggtcaata
     3601 atcaggaagt gatggagcat cagggcggct atacgccatt tgaagccgat gtcacgccgt
     3661 atgttattgc cgggaaaagt gtacgtatca ccgtttgtgt gaacaacgaa ctgaactggc
     3721 agactatccc gccgggaatg gtgattaccg acgaaaacgg caagaaaaag cagtcttact
     3781 tccatgattt ctttaactat gccggaatcc atcgcagcgt aatgctctac accacgccga
     3841 acacctgggt ggacgatatc accgtggtga cgcatgtcgc gcaagactgt aaccacgcgt
     3901 ctgttgactg gcaggtggtg gccaatggtg atgtcagcgt tgaactgcgt gatgcggatc
     3961 aacaggtggt tgcaactgga caaggcacta gcgggacttt gcaagtggtg aatccgcacc
     4021 tctggcaacc gggtgaaggt tatctctatg aactgtgcgt cacagccaaa agccagacag
     4081 agtgtgatat ctacccgctt cgcgtcggca tccggtcagt ggcagtgaag ggcgaacagt
     4141 tcctgattaa ccacaaaccg ttctacttta ctggctttgg tcgtcatgaa gatgcggact
     4201 tacgtggcaa aggattcgat aacgtgctga tggtgcacga ccacgcatta atggactgga
     4261 ttggggccaa ctcctaccgt acctcgcatt acccttacgc tgaagagatg ctcgactggg
     4321 cagatgaaca tggcatcgtg gtgattgatg aaactgctgc tgtcggcttt aacctctctt
     4381 taggcattgg tttcgaagcg ggcaacaagc cgaaagaact gtacagcgaa gaggcagtca
     4441 acggggaaac tcagcaagcg cacttacagg cgattaaaga gctgatagcg cgtgacaaaa
     4501 accacccaag cgtggtgatg tggagtattg ccaacgaacc ggatacccgt ccgcaaggtg
     4561 cacgggaata tttcgcgcca ctggcggaag caacgcgtaa actcgacccg acgcgtccga
     4621 tcacctgcgt caatgtaatg ttctgcgacg ctcacaccga taccatcagc gatctctttg
     4681 atgtgctgtg cctgaaccgt tattacggat ggtatgtcca aagcggcgat ttggaaacgg
     4741 cagagaaggt actggaaaaa gaacttctgg cctggcagga gaaactgcat cagccgatta
     4801 tcatcaccga atacggcgtg gatacgttag ccgggctgca ctcaatgtac accgacatgt
     4861 ggagtgaaga gtatcagtgt gcatggctgg atatgtatca ccgcgtcttt gatcgcgtca
     4921 gcgccgtcgt cggtgaacag gtatggaatt tcgccgattt tgcgacctcg caaggcatat
     4981 tgcgcgttgg cggtaacaag aaagggatct tcactcgcga ccgcaaaccg aagtcggcgg
     5041 cttttctgct gcaaaaacgc tggactggca tgaacttcgg tgaaaaaccg cagcagggag
     5101 gcaaacaatg acacccagct tt
//
