LOCUS       sequence-377981-        8577 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To be a control plasmid that has no Rluc repression (replaced by 
            turboGFP) and should reflect true off state of the circuit..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8577)
  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 8577)
  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: LLP726_L2_pV01_0I_TMV-tGFP_TCTP-fLUC
FEATURES             Location/Qualifiers
     source          1..8577
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    824..879
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             complement(1569..1586)
                     /codon_start=1
                     /product="HQHQHQ metal affinity tag"
                     /label=HQ tag
                     /note="The HQ tag binds to nickel and other metal ions much
                     like the 6xHis tag, but may allow elution at lower 
                     concentrations of imidazole (Godat et al., 2008)."
                     /translation="HQHQHQ"
     polyA_signal    1857..2031
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             2419..4071
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(2485..2503)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     terminator      4091..4291
                     /label=gene 7 terminator
                     /note="transcription terminator for octopine-type Ti 
                     plasmid gene 7"
     misc_feature    4355..4379
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      4467..5177
                     /label=oriV
                     /note="incP origin of replication"
     promoter        5198..5302
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5303..6097
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      6268..6856
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6757..6776
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             7192..8340
                     /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    8486..8510
                     /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 caaacaacaa acaacattac aattactatt tacaattaca atgagaggat ctggatctga
      901 gtctgatgag tctggacttc ctgctatgga aatcgagtgt agaatcactg gaacccttaa
      961 cggtgttgag ttcgagcttg ttggaggtgg tgagggaact cctgagcagg gaagaatgac
     1021 taacaagatg aagtctacca agggtgctct taccttctct ccataccttc tttctcacgt
     1081 tatgggatac ggattctacc acttcggaac ttacccatct ggatacgaga accctttcct
     1141 tcatgctatc aacaacggtg gatacaccaa cactaggatc gagaagtacg aggatggtgg
     1201 tgttcttcac gttagcttct cttacagata cgaggctgga agagtgatcg gagatttcaa
     1261 ggttatggga actggattcc ctgaggattc tgttatcttc accgacaaga tcatcaggtc
     1321 taacgctact gttgagcatc ttcatcctat gggagataac gatctcgatg gatctttcac
     1381 cagaaccttc tcacttagag atggtggtta ctactcttct gtggtggatt ctcacatgca
     1441 cttcaagtct gctatccacc cttctatcct tcaaaacggt ggacctatgt tcgctttcag
     1501 aagagttgag gaagatcact ctaacaccga gcttggaatc gttgagtacc aacatgcttt
     1561 caagacccct gatgctgatg ctggtgagga attcgaagag caagcaagga aagctaaggt
     1621 gaacaatgag aaaaagacgg aaatagtgaa accagagagt tgtagcaatg aaggagatgt
     1681 caaggatctg aaaagaaagg actctgagga tggaaacgag ggtgaggaag aagaagcttc
     1741 ttcgaaaccg aaaaagccaa aagttgctct ttctcatctt caggacattg acgacacaga
     1801 agctgatcaa gaagaagagt aagcttctct agctagagtc gatcgacaag ctcgagtttc
     1861 tccataataa tgtgtgagta gttcccagat aagggaatta gggttcctat agggtttcgc
     1921 tcatgtgttg agcatataag aaacccttag tatgtatttg tatttgtaaa atacttctat
     1981 caataaaatt tctaattcct aaaaccaaaa tccagtacta aaatccagat cgctgcaaga
     2041 attcaagctt ggagccaaca ctcgaatccc cacccgttgg accaaacccg gctcattaag
     2101 cgtcggttca gatttatttc ctttatttaa aaaaaaggaa agggtaaaaa atagaaaatt
     2161 ggaaacagtt aaagcccaaa attgtaattt accgagaatt gtaaatttac ctgaaaaccc
     2221 tacgctatag tttcgactat aaataccaaa cttaggacct cacttcagaa tcccctcgtc
     2281 gctgcgtctc tctcccgcaa ccttcgattt tcgtttattc gcatccatcg gagagagaaa
     2341 acaatcaata agcgacctac tacaaacaag taagtaagag aaaaaccaaa agaagaagag
     2401 aaacaacaag aagaagtaat ggaggacgcc aaaaacataa agaaaggccc ggcgccattc
     2461 tatcctctag aggatggaac cgctggagag caactgcata aggctatgaa gagatacgcc
     2521 ctggttcctg gaacaattgc ttttacagat gcacatatcg aggtgaacat cacgtacgcg
     2581 gaatacttcg aaatgtccgt tcggttggca gaagctatga aacgatatgg gctgaataca
     2641 aatcacagaa tcgtcgtatg cagtgaaaac tctcttcaat tctttatgcc ggtgttgggc
     2701 gcgttattta tcggagttgc agttgcgccc gcgaacgaca tttataatga acgtgaattg
     2761 ctcaacagta tgaacatttc gcagcctacc gtagtgtttg tttccaaaaa ggggttgcaa
     2821 aaaattttga acgtgcaaaa aaaattacca ataatccaga aaattattat catggattct
     2881 aaaacggatt accagggatt tcagtcgatg tacacgttcg tcacatctca tctacctccc
     2941 ggttttaatg aatacgattt tgtaccagag tcctttgatc gtgacaaaac aattgcactg
     3001 ataatgaatt cctctggatc tactgggtta cctaagggtg tggcccttcc gcatagaact
     3061 gcctgcgtca gattctcgca tgccagagat cctatttttg gcaatcaaat cattccggat
     3121 actgcgattt taagtgttgt tccattccat cacggttttg gaatgtttac tacactcgga
     3181 tatttgatat gtggatttcg agtcgtctta atgtatagat ttgaagaaga gctgttttta
     3241 cgatcccttc aggattacaa aattcaaagt gcgttgctag taccaaccct attttcattc
     3301 ttcgccaaaa gcactctgat tgacaaatac gatttatcta atttacacga aattgcttct
     3361 gggggcgcac ctctttcgaa agaagtcggg gaagcggttg caaaacgctt ccatcttcca
     3421 gggatacgac aaggatatgg gctcactgag actacatcag ctattctgat tacacccgag
     3481 ggggatgata aaccgggcgc ggtcggtaaa gttgttccat tttttgaagc gaaggttgtg
     3541 gatctggata ccgggaaaac gctgggcgtt aatcagagag gcgaattatg tgtcagagga
     3601 cctatgatta tgtccggtta tgtaaacaat ccggaagcga ccaacgcctt gattgacaag
     3661 gatggatggc tacattctgg agacatagct tactgggacg aggacgaaca cttcttcata
     3721 gttgaccgct tgaagtcttt aattaaatac aaaggatatc aggtggcccc cgctgaattg
     3781 gaatcgatat tgttacaaca ccccaacatc ttcgacgcgg gcgtggcagg tctgcccgac
     3841 gatgacgccg gtgaacttcc cgccgccgtt gttgttttgg agcacggaaa gacgatgacg
     3901 gaaaaagaga tcgtggatta cgtcgccagt caagtaacaa ccgcgaaaaa gttgcgcgga
     3961 ggagttgtgt ttgtggacga agtaccgaaa ggtcttaccg gaaaactcga cgcaagaaaa
     4021 atcagagaga tcctcataaa ggccaagaag ggcggaaagt ccaaattgta agcttgatcc
     4081 cccgtcgaca gctagctata tcatcaattt atgtattaca cataatatcg cactcagtct
     4141 ttcatctacg gcaatgtacc agctgatata atcagttatt gaaatatttc tgaatttaaa
     4201 cttgcatcaa taaatttatg tttttgcttg gactataata cctgacttgt tattttatca
     4261 ataaatattt aaactatatt tctttcaaga tcgctactag aggatgcaca tgtgaccgag
     4321 ggacacgaag tgatccgttt aaactatcag tgtttgacag gatatattgg cgggtaaacc
     4381 taagagaaaa gagcgtttat tagaataatc ggatatttaa aagggcgtga aaaggtttat
     4441 ccgttcgtcc atttgtatgt gccagccgcc tttgcgacgc tcaccgggct ggttgccctc
     4501 gccgctgggc tggcggccgt ctatggccct gcaaacgcgc cagaaacgcc gtcgaagccg
     4561 tgtgcgagac accgcggccg ccggcgttgt ggatacctcg cggaaaactt ggccctcact
     4621 gacagatgag gggcggacgt tgacacttga ggggccgact cacccggcgc ggcgttgaca
     4681 gatgaggggc aggctcgatt tcggccggcg acgtggagct ggccagcctc gcaaatcggc
     4741 gaaaacgcct gattttacgc gagtttccca cagatgatgt ggacaagcct ggggataagt
     4801 gccctgcggt attgacactt gaggggcgcg actactgaca gatgaggggc gcgatccttg
     4861 acacttgagg ggcagagtgc tgacagatga ggggcgcacc tattgacatt tgaggggctg
     4921 tccacaggca gaaaatccag catttgcaag ggtttccgcc cgtttttcgg ccaccgctaa
     4981 cctgtctttt aacctgcttt taaaccaata tttataaacc ttgtttttaa ccagggctgc
     5041 gccctgtgcg cgtgaccgcg cacgccgaag gggggtgccc ccccttctcg aaccctcccg
     5101 gcccgctaac gcgggcctcc catcccccca ggggctgcgc ccctcggccg cgaacggcct
     5161 caccccaaaa atggcagcgc tggccaattc ccgagtgcgc ggaaccccta tttgtttatt
     5221 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5281 ataatattga aaaaggaaga gtatggctaa aatgagaata tcaccggaat tgaaaaaact
     5341 gatcgaaaaa taccgctgcg taaaagatac ggaaggaatg tctcctgcta aggtatataa
     5401 gctggtggga gaaaatgaaa acctatattt aaaaatgacg gacagccggt ataaagggac
     5461 cacctatgat gtggaacggg aaaaggacat gatgctatgg ctggaaggaa agctgcctgt
     5521 tccaaaggtc ctgcactttg aacggcatga tggctggagc aatctgctca tgagtgaggc
     5581 cgatggcgtc ctttgctcgg aagagtatga agatgaacaa agccctgaaa agattatcga
     5641 gctgtatgcg gagtgcatca ggctctttca ctccatcgac atatcggatt gtccctatac
     5701 gaatagctta gacagccgct tagccgaatt ggattactta ctgaataacg atctggccga
     5761 tgtggattgc gaaaactggg aagaggacac tccatttaaa gatccgcgcg agctgtatga
     5821 ttttttaaag acggaaaagc ccgaagagga acttgtcttt tcccacggcg acctgggaga
     5881 cagcaacatc tttgtgaaag atggcaaagt aagtggcttt attgatcttg ggagaagcgg
     5941 cagggcggac aagtggtatg acattgcctt ctgcgtccgg tcgctcaggg aggatatcgg
     6001 ggaagaacag tatgtcgagc tattttttga cttactgggg atcaagcctg attgggagaa
     6061 aataaaatat tatattttac tggatgaatt gttttagctg tcagaccaag tttactcata
     6121 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     6181 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6241 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6301 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6361 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6421 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6481 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6541 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6601 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6661 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6721 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6781 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6841 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg cagatcctag
     6901 atgtggcgca acgatgccgg cgacaagcag gagcgcaccg acttcttccg catcaagtgt
     6961 tttggctctc aggccgaggc ccacggcaag tatttgggca aggggtcgct ggtattcgtg
     7021 cagggcaaga ttcggaatac caagtacgag aaggacggcc agacggtcta cgggaccgac
     7081 ttcattgccg ataaggtgga ttatctggac accaaggcac caggcgggtc aaatcaggaa
     7141 taagggcaca ttgccccggc gtgagtcggg gcaatcccgc aaggagggtg aatgaatcgg
     7201 acgtttgacc ggaaggcata caggcaagaa ctgatcgacg cggggttttc cgccgaggat
     7261 gccgaaacca tcgcaagccg caccgtcatg cgtgcgcccc gcgaaacctt ccagtccgtc
     7321 ggctcgatgg tccagcaagc tacggccaag atcgagcgcg acagcgtgca actggctccc
     7381 cctgccctgc ccgcgccatc ggccgccgtg gagcgttcgc gtcgtcttga acaggaggcg
     7441 gcaggtttgg cgaagtcgat gaccatcgac acgcgaggaa ctatgacgac caagaagcga
     7501 aaaaccgccg gcgaggacct ggcaaaacag gtcagcgagg ccaagcaggc cgcgttgctg
     7561 aaacacacga agcagcagat caaggaaatg cagctttcct tgttcgatat tgcgccgtgg
     7621 ccggacacga tgcgagcgat gccaaacgac acggcccgct ctgccctgtt caccacgcgc
     7681 aacaagaaaa tcccgcgcga ggcgctgcaa aacaaggtca ttttccacgt caacaaggac
     7741 gtgaagatca cctacaccgg cgtcgagctg cgggccgacg atgacgaact ggtgtggcag
     7801 caggtgttgg agtacgcgaa gcgcacccct atcggcgagc cgatcacctt cacgttctac
     7861 gagctttgcc aggacctggg ctggtcgatc aatggccggt attacacgaa ggccgaggaa
     7921 tgcctgtcgc gcctacaggc gacggcgatg ggcttcacgt ccgaccgcgt tgggcacctg
     7981 gaatcggtgt cgctgctgca ccgcttccgc gtcctggacc gtggcaagaa aacgtcccgt
     8041 tgccaggtcc tgatcgacga ggaaatcgtc gtgctgtttg ctggcgacca ctacacgaaa
     8101 ttcatatggg agaagtaccg caagctgtcg ccgacggccc gacggatgtt cgactatttc
     8161 agctcgcacc gggagccgta cccgctcaag ctggaaacct tccgcctcat gtgcggatcg
     8221 gattccaccc gcgtgaagaa gtggcgcgag caggtcggcg aagcctgcga agagttgcga
     8281 ggcagcggcc tggtggaaca cgcctgggtc aatgatgacc tggtgcattg caaacgctag
     8341 ggccttgtgg ggtcagttcc ggctgggggt tcagcagccc ctgctcggat ctgttggacc
     8401 ggacagtagt catggttgat gggctgcctg tatcgagtgg tgattttgtg ccgagctgcc
     8461 ggtcggggag ctgttggctg gctggtggca ggatatattg tggtgtaaac aaattgacgc
     8521 ttagacaact taataacaca ttgcggacgt ttttaatgta ctggggttga acactct
//
