LOCUS       sequence-378263-        9142 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To test for the repressive potential of the conserved peptide uORF 
            from AT1G36730 with the FRT sites around it..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9142)
  AUTHORS   Lloyd JPB, Ly F, Gong P, Pflueger J, Swain T, Pflueger C, Fourie E, 
            Khan MA, Kidd BN, Lister R
  TITLE     Synthetic memory circuits for stable cell reprogramming in plants.
  JOURNAL   Nat Biotechnol. 2022 Jul 4. pii: 10.1038/s41587-022-01383-2. doi: 
            10.1038/s41587-022-01383-2.
  PUBMED    35788565
REFERENCE   2  (bases 1 to 9142)
  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: "Nat 
            Biotechnol. 2022 Jul 4. pii: 10.1038/s41587-022-01383-2. doi: 
            10.1038/s41587-022-01383-2."
COMMENT     Sequence Label: LLP733_L2_pV01_0I_FRT-uORF-rLUC_TCTP-fLUC
FEATURES             Location/Qualifiers
     source          1..9142
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    845..878
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    1106..1139
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     intron          1206..1394
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     polyA_signal    2422..2596
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             2984..4636
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(3050..3068)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     terminator      4656..4856
                     /label=gene 7 terminator
                     /note="transcription terminator for octopine-type Ti 
                     plasmid gene 7"
     misc_feature    4920..4944
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      5032..5742
                     /label=oriV
                     /note="incP origin of replication"
     promoter        5763..5867
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5868..6662
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      6833..7421
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7322..7341
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             7757..8905
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     misc_feature    9051..9075
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 gtgccgaatt cggatccgga gtcgacaaaa tttagaacga acttaattat gatctcaaat
       61 acattgatac atatctcatc tagatctagg ttatcattat gtaagaaagt tttgacgaat
      121 atggcacgac aaaatggcta gactcgatgt aattggtatc tcaactcaac attatactta
      181 taccaaacat tagttagaca aaatttaaac aactattttt tatgtatgca agagtcagca
      241 tatgtataat tgattcagaa tcgttttgac gagttcggat gtagtagtag ccattattta
      301 atgtacatac taatcgtgaa tagtgaatat gatgaaacat tgtatcttat tgtataaata
      361 tccataaaca catcatgaaa gacactttct ttcacggtct gaattaatta tgatacaatt
      421 ctaatagaaa acgaattaaa ttacgttgaa ttgtatgaaa tctaattgaa caagccaacc
      481 acgacgacga ctaacgttgc ctggattgac tcggtttaag ttaaccacta aaaaaacgga
      541 gctgtcatgt aacacgcgga tcgagcaggt cacagtcatg aagccatcaa agcaaaagaa
      601 ctaatccaag ggctgagatg attaattagt ttaaaaatta gttaacacga gggaaaaggc
      661 tgtctgacag ccaggtcacg ttatctttac ctgtggtcga aatgattcgt gtctgtcgat
      721 tttaattatt tttttgaaag gccgaaaata aagttgtaag agataaaccc gcctatataa
      781 attcatatat tttcctctcc gctttgaata ctgtattttt acaacaatta ccaacaacaa
      841 caaagaagtt cctattctct agaaagtata ggaacttcaa taaagaatct tttatgtctg
      901 aacaggcttc tctctttcaa tcatctatcc ggcttttagg agagtactgt cctatatcta
      961 ggaaaaagtc cactcataaa gttggtggtt tgcgttttga ggaggacatc acgagctgta
     1021 ttggatttca gtttatgcaa ctgagtgatt atcactgcct cttctgacat cccttgtttg
     1081 tttacgtttg ttgaatcaag actctgaagt tcctattctc tagaaagtat aggaacttcc
     1141 aacaaacaac attacaatta ctatttacaa ttacaatgac ttcaaaggtg tatgatcctg
     1201 agcaggtaag tttctgcttc tacctttgat atatatataa taattatcat taattagtag
     1261 taatataata tttcaaatat ttttttcaaa ataaaagaat gtagtatata gcaattgctt
     1321 ttctgtagtt tataagtgtg tatattttaa tttataactt ttctaatata tgaccaaaat
     1381 ttgttgatgt gcagagaaaa agaatgataa ctggtccaca atggtgggca agatgtaaac
     1441 agatgaacgt gcttgattct ttcattaatt actacgattc agaaaagcac gctgagaatg
     1501 cagttatctt tcttcacgga aacgctgcat cttcatattt gtggagacat gttgtgcctc
     1561 acattgaacc agtggctagg tgtattatcc cagatcttat cggaatggga aagtctggta
     1621 aatctggaaa tggttcttac agacttttgg atcattataa gtacctcaca gcttggttcg
     1681 aacttctcaa cttacctaag aagattattt tcgttggaca cgattggggt gcttgcttgg
     1741 cattccatta ctcatacgaa caccaagata agatcaaagc tatagttcat gcagagagtg
     1801 ttgtggatgt gatagaatct tgggatgagt ggccagatat tgaagaggat attgctttga
     1861 tcaagtctga agagggagaa aagatggttc tcgaaaataa ctttttcgtg gaaactatgc
     1921 ttccttcaaa gattatgaga aaattggaac cagaagagtt tgctgcatat ctcgaacctt
     1981 tcaaggaaaa aggagaggtt agaaggccaa cactcagttg gcctagggaa ataccattag
     2041 ttaagggagg taaacctgat gttgtgcaga tcgtgagaaa ttataacgct taccttaggg
     2101 catcagatga tttgcctaag atgttcatcg agtcagatcc aggatttttc agtaatgcta
     2161 tagttgaagg tgcaaagaaa tttccaaaca ccgagttcgt taaggtgaaa ggattgcatt
     2221 tctcacaaga agatgctcct gatgagatgg gaaagtatat taagtctttt gttgaaagag
     2281 tgctcaaaaa cgagcagggt tctcgatctg atgcttctga agtaacgtca acaagctccc
     2341 agtctgaggt atgcacagtg gaaacgccag gttgtgttgg ttcttaagct tctctagcta
     2401 gagtcgatcg acaagctcga gtttctccat aataatgtgt gagtagttcc cagataaggg
     2461 aattagggtt cctatagggt ttcgctcatg tgttgagcat ataagaaacc cttagtatgt
     2521 atttgtattt gtaaaatact tctatcaata aaatttctaa ttcctaaaac caaaatccag
     2581 tactaaaatc cagatcgctg caagaattca agcttggagc caacactcga atccccaccc
     2641 gttggaccaa acccggctca ttaagcgtcg gttcagattt atttccttta tttaaaaaaa
     2701 aggaaagggt aaaaaataga aaattggaaa cagttaaagc ccaaaattgt aatttaccga
     2761 gaattgtaaa tttacctgaa aaccctacgc tatagtttcg actataaata ccaaacttag
     2821 gacctcactt cagaatcccc tcgtcgctgc gtctctctcc cgcaaccttc gattttcgtt
     2881 tattcgcatc catcggagag agaaaacaat caataagcga cctactacaa acaagtaagt
     2941 aagagaaaaa ccaaaagaag aagagaaaca acaagaagaa gtaatggagg acgccaaaaa
     3001 cataaagaaa ggcccggcgc cattctatcc tctagaggat ggaaccgctg gagagcaact
     3061 gcataaggct atgaagagat acgccctggt tcctggaaca attgctttta cagatgcaca
     3121 tatcgaggtg aacatcacgt acgcggaata cttcgaaatg tccgttcggt tggcagaagc
     3181 tatgaaacga tatgggctga atacaaatca cagaatcgtc gtatgcagtg aaaactctct
     3241 tcaattcttt atgccggtgt tgggcgcgtt atttatcgga gttgcagttg cgcccgcgaa
     3301 cgacatttat aatgaacgtg aattgctcaa cagtatgaac atttcgcagc ctaccgtagt
     3361 gtttgtttcc aaaaaggggt tgcaaaaaat tttgaacgtg caaaaaaaat taccaataat
     3421 ccagaaaatt attatcatgg attctaaaac ggattaccag ggatttcagt cgatgtacac
     3481 gttcgtcaca tctcatctac ctcccggttt taatgaatac gattttgtac cagagtcctt
     3541 tgatcgtgac aaaacaattg cactgataat gaattcctct ggatctactg ggttacctaa
     3601 gggtgtggcc cttccgcata gaactgcctg cgtcagattc tcgcatgcca gagatcctat
     3661 ttttggcaat caaatcattc cggatactgc gattttaagt gttgttccat tccatcacgg
     3721 ttttggaatg tttactacac tcggatattt gatatgtgga tttcgagtcg tcttaatgta
     3781 tagatttgaa gaagagctgt ttttacgatc ccttcaggat tacaaaattc aaagtgcgtt
     3841 gctagtacca accctatttt cattcttcgc caaaagcact ctgattgaca aatacgattt
     3901 atctaattta cacgaaattg cttctggggg cgcacctctt tcgaaagaag tcggggaagc
     3961 ggttgcaaaa cgcttccatc ttccagggat acgacaagga tatgggctca ctgagactac
     4021 atcagctatt ctgattacac ccgaggggga tgataaaccg ggcgcggtcg gtaaagttgt
     4081 tccatttttt gaagcgaagg ttgtggatct ggataccggg aaaacgctgg gcgttaatca
     4141 gagaggcgaa ttatgtgtca gaggacctat gattatgtcc ggttatgtaa acaatccgga
     4201 agcgaccaac gccttgattg acaaggatgg atggctacat tctggagaca tagcttactg
     4261 ggacgaggac gaacacttct tcatagttga ccgcttgaag tctttaatta aatacaaagg
     4321 atatcaggtg gcccccgctg aattggaatc gatattgtta caacacccca acatcttcga
     4381 cgcgggcgtg gcaggtctgc ccgacgatga cgccggtgaa cttcccgccg ccgttgttgt
     4441 tttggagcac ggaaagacga tgacggaaaa agagatcgtg gattacgtcg ccagtcaagt
     4501 aacaaccgcg aaaaagttgc gcggaggagt tgtgtttgtg gacgaagtac cgaaaggtct
     4561 taccggaaaa ctcgacgcaa gaaaaatcag agagatcctc ataaaggcca agaagggcgg
     4621 aaagtccaaa ttgtaagctt gatcccccgt cgacagctag ctatatcatc aatttatgta
     4681 ttacacataa tatcgcactc agtctttcat ctacggcaat gtaccagctg atataatcag
     4741 ttattgaaat atttctgaat ttaaacttgc atcaataaat ttatgttttt gcttggacta
     4801 taatacctga cttgttattt tatcaataaa tatttaaact atatttcttt caagatcgct
     4861 actagaggat gcacatgtga ccgagggaca cgaagtgatc cgtttaaact atcagtgttt
     4921 gacaggatat attggcgggt aaacctaaga gaaaagagcg tttattagaa taatcggata
     4981 tttaaaaggg cgtgaaaagg tttatccgtt cgtccatttg tatgtgccag ccgcctttgc
     5041 gacgctcacc gggctggttg ccctcgccgc tgggctggcg gccgtctatg gccctgcaaa
     5101 cgcgccagaa acgccgtcga agccgtgtgc gagacaccgc ggccgccggc gttgtggata
     5161 cctcgcggaa aacttggccc tcactgacag atgaggggcg gacgttgaca cttgaggggc
     5221 cgactcaccc ggcgcggcgt tgacagatga ggggcaggct cgatttcggc cggcgacgtg
     5281 gagctggcca gcctcgcaaa tcggcgaaaa cgcctgattt tacgcgagtt tcccacagat
     5341 gatgtggaca agcctgggga taagtgccct gcggtattga cacttgaggg gcgcgactac
     5401 tgacagatga ggggcgcgat ccttgacact tgaggggcag agtgctgaca gatgaggggc
     5461 gcacctattg acatttgagg ggctgtccac aggcagaaaa tccagcattt gcaagggttt
     5521 ccgcccgttt ttcggccacc gctaacctgt cttttaacct gcttttaaac caatatttat
     5581 aaaccttgtt tttaaccagg gctgcgccct gtgcgcgtga ccgcgcacgc cgaagggggg
     5641 tgccccccct tctcgaaccc tcccggcccg ctaacgcggg cctcccatcc ccccaggggc
     5701 tgcgcccctc ggccgcgaac ggcctcaccc caaaaatggc agcgctggcc aattcccgag
     5761 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     5821 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg gctaaaatga
     5881 gaatatcacc ggaattgaaa aaactgatcg aaaaataccg ctgcgtaaaa gatacggaag
     5941 gaatgtctcc tgctaaggta tataagctgg tgggagaaaa tgaaaaccta tatttaaaaa
     6001 tgacggacag ccggtataaa gggaccacct atgatgtgga acgggaaaag gacatgatgc
     6061 tatggctgga aggaaagctg cctgttccaa aggtcctgca ctttgaacgg catgatggct
     6121 ggagcaatct gctcatgagt gaggccgatg gcgtcctttg ctcggaagag tatgaagatg
     6181 aacaaagccc tgaaaagatt atcgagctgt atgcggagtg catcaggctc tttcactcca
     6241 tcgacatatc ggattgtccc tatacgaata gcttagacag ccgcttagcc gaattggatt
     6301 acttactgaa taacgatctg gccgatgtgg attgcgaaaa ctgggaagag gacactccat
     6361 ttaaagatcc gcgcgagctg tatgattttt taaagacgga aaagcccgaa gaggaacttg
     6421 tcttttccca cggcgacctg ggagacagca acatctttgt gaaagatggc aaagtaagtg
     6481 gctttattga tcttgggaga agcggcaggg cggacaagtg gtatgacatt gccttctgcg
     6541 tccggtcgct cagggaggat atcggggaag aacagtatgt cgagctattt tttgacttac
     6601 tggggatcaa gcctgattgg gagaaaataa aatattatat tttactggat gaattgtttt
     6661 agctgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     6721 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6781 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6841 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6901 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6961 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     7021 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     7081 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     7141 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     7201 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     7261 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     7321 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     7381 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     7441 ttttacggtt cctggcagat cctagatgtg gcgcaacgat gccggcgaca agcaggagcg
     7501 caccgacttc ttccgcatca agtgttttgg ctctcaggcc gaggcccacg gcaagtattt
     7561 gggcaagggg tcgctggtat tcgtgcaggg caagattcgg aataccaagt acgagaagga
     7621 cggccagacg gtctacggga ccgacttcat tgccgataag gtggattatc tggacaccaa
     7681 ggcaccaggc gggtcaaatc aggaataagg gcacattgcc ccggcgtgag tcggggcaat
     7741 cccgcaagga gggtgaatga atcggacgtt tgaccggaag gcatacaggc aagaactgat
     7801 cgacgcgggg ttttccgccg aggatgccga aaccatcgca agccgcaccg tcatgcgtgc
     7861 gccccgcgaa accttccagt ccgtcggctc gatggtccag caagctacgg ccaagatcga
     7921 gcgcgacagc gtgcaactgg ctccccctgc cctgcccgcg ccatcggccg ccgtggagcg
     7981 ttcgcgtcgt cttgaacagg aggcggcagg tttggcgaag tcgatgacca tcgacacgcg
     8041 aggaactatg acgaccaaga agcgaaaaac cgccggcgag gacctggcaa aacaggtcag
     8101 cgaggccaag caggccgcgt tgctgaaaca cacgaagcag cagatcaagg aaatgcagct
     8161 ttccttgttc gatattgcgc cgtggccgga cacgatgcga gcgatgccaa acgacacggc
     8221 ccgctctgcc ctgttcacca cgcgcaacaa gaaaatcccg cgcgaggcgc tgcaaaacaa
     8281 ggtcattttc cacgtcaaca aggacgtgaa gatcacctac accggcgtcg agctgcgggc
     8341 cgacgatgac gaactggtgt ggcagcaggt gttggagtac gcgaagcgca cccctatcgg
     8401 cgagccgatc accttcacgt tctacgagct ttgccaggac ctgggctggt cgatcaatgg
     8461 ccggtattac acgaaggccg aggaatgcct gtcgcgccta caggcgacgg cgatgggctt
     8521 cacgtccgac cgcgttgggc acctggaatc ggtgtcgctg ctgcaccgct tccgcgtcct
     8581 ggaccgtggc aagaaaacgt cccgttgcca ggtcctgatc gacgaggaaa tcgtcgtgct
     8641 gtttgctggc gaccactaca cgaaattcat atgggagaag taccgcaagc tgtcgccgac
     8701 ggcccgacgg atgttcgact atttcagctc gcaccgggag ccgtacccgc tcaagctgga
     8761 aaccttccgc ctcatgtgcg gatcggattc cacccgcgtg aagaagtggc gcgagcaggt
     8821 cggcgaagcc tgcgaagagt tgcgaggcag cggcctggtg gaacacgcct gggtcaatga
     8881 tgacctggtg cattgcaaac gctagggcct tgtggggtca gttccggctg ggggttcagc
     8941 agcccctgct cggatctgtt ggaccggaca gtagtcatgg ttgatgggct gcctgtatcg
     9001 agtggtgatt ttgtgccgag ctgccggtcg gggagctgtt ggctggctgg tggcaggata
     9061 tattgtggtg taaacaaatt gacgcttaga caacttaata acacattgcg gacgttttta
     9121 atgtactggg gttgaacact ct
//
