LOCUS       sequence-129830-        3897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry clone containing 3AT. Necessary to complete the 
            transcriptional reporters by providing the poly-A tail. 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 3897)
  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 3897)
  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: p2R3a-3AT
FEATURES             Location/Qualifiers
     source          1..3897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(126..144)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(149..165)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(246..264)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(247..264)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(282..298)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(295..317)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    306..322
                     /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(330..336,337..354,355..360))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    375..396
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(513..530)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(684..1272)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(764..783)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1443..2234,2235..2303))
                     /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     2066..2085
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2304..2408)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2486..2941)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2623..2644)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2835..2854
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3067..3089
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3082..3098
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3105..3123
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3154..3163
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3158..3167
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     3184..3200
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3271..3394)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(join(3890..3897,1..88))
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ttatacaaag ttggcattat aaaaaagcat tgcttatcaa tttgttgcaa cgaacaggtc
       61 actatcagtc aaaataaaat cattatttgg agctccatgg tagcgttaac gcggccgcga
      121 tatcccctat agtgagtcgt attacatggt catagctgtt tcctgaatca ctagtgaatt
      181 cgcggccgcc tgcaggtcga ccatatggga gagctcccaa cgcgttggat gcatagcttg
      241 agtattctat agtgtcacct aaatagcttg gcgtaatcat ggtcatagct gtttcctgtg
      301 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
      361 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
      421 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
      481 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
      541 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
      601 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
      661 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
      721 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
      781 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
      841 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
      901 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
      961 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     1021 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     1081 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     1141 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1201 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1261 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     1321 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     1381 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     1441 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     1501 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     1561 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     1621 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     1681 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     1741 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     1801 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     1861 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     1921 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     1981 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     2041 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     2101 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     2161 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     2221 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     2281 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     2341 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     2401 gttccgcgca catttccccg aaaagtgcca cctgatgcgg tgtgaaatac cgcacagatg
     2461 cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg
     2521 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     2581 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     2641 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     2701 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     2761 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     2821 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     2881 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     2941 tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     3001 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     3061 ggttttccca gtcacgacgt tgtaaaacga cggccagtga attgtaatac gactcactat
     3121 agggcgaatt gggcccgacg tcgcatgctc ccggccgcca tggcggccgc gggaattcga
     3181 tttgtaaaac gacggccagt cttaagctcg ggccctgcag ctctagagct cgaattctac
     3241 aggtcactaa taccatctaa gtagttggtt catagtgact gcatatgttg tgttttacag
     3301 tattatgtag tctgtttttt atgcaaaatc taatttaata tattgatatt tatatcattt
     3361 tacgtttctc gttcaacttt cttgtacaaa gtggaataac aggcctccca gctttcgtcc
     3421 gtatcatcgg tttcgacaac gttcgtcaag ttcaatgcat cagtttcatt gcccacacac
     3481 cagaatccta ctaagtttga gtattatggc attggaaaag ctgttttctt ctatcatttg
     3541 ttctgcttgt aatttactgt gttctttcag tttttgtttt cggacatcaa aatgcaaatg
     3601 gatggataag agttaataaa tgatatggtc cttttgttca ttctcaaatt attattatct
     3661 gttgttttta ctttaatggg ttgaatttaa gtaagaaagg aactaacagt gtgatattaa
     3721 ggtgcaatgt tagacatata aaacagtctt tcacctctct ttggttatgt cttgaattgg
     3781 tttgtttctt cacttatctg tgtaatcaag tttactatga gtctatgatc aagtaattat
     3841 gcaatcaagt taagtacagt ataggctttt tgtgtcgagg gtttaaacac aacttta
//
