LOCUS       sequence-387027-        5720 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proG1090::LmrA-VP16-19St).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5720)
  AUTHORS   Brophy JAN, Magallon KJ, Duan L, Zhong V, Ramachandran P, Kniazev K,
            Dinneny JR
  TITLE     Synthetic genetic circuits as a means of reprogramming plant roots.
  JOURNAL   Science. 2022 Aug 12;377(6607):747-751. doi: 
            10.1126/science.abo4326. Epub 2022 Aug 11.
  PUBMED    35951698
REFERENCE   2  (bases 1 to 5720)
  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: 
            "10.1126/science.abo4326"; journalName: "Science"; date: 
            "2022-08-12- 12"; volume: "377"; issue: "6607"; pages: "747-751"
COMMENT     Sequence Label: JAB2164
FEATURES             Location/Qualifiers
     source          1..5720
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    8..32
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(102..354)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(374..925)
                     /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(967..1146)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     1463..1483
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        1472..1518
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             2101..2334
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDTADFEFEQMFTDALGIDEYGG"
     CDS             2335..2355
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2411..2587
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(2669..2689)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2669..2687)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2708..2724)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2708..2724)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2721..2743)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2732..2748
                     /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(2756..2762,2763..2780,2781..2786))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2801..2822
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2939..2956)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3061..3085
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(3176..3764)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3256..3275)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3855..4643)
                     /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      5101..5536
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 tggctggtgg caggatatat tgtggtgtaa acaaattgac gcttagacaa cttaataaca
       61 cattgcggac gtttttaatg taatcagctt gcatgccggt cgatctagta acatagatga
      121 caccgcgcgc gataatttat cctagtttgc gcgctatatt ttgttttcta tcgcgtatta
      181 aatgtataat tgcgggactc taatcataaa aacccatctc ataaataacg tcatgcatta
      241 catgttaatt attacatgct taacgtaatt caacagaaat tatatgataa tcatcgcaag
      301 accggcaaca ggattcaatc ttaagaaact ttattgccaa atgtttgaac gatctgcttg
      361 actctagggg tcatcagatt tcggtgacgg gcaggaccgg acggggcggc accggcaggc
      421 tgaagtccag ctgccagaaa cccacgtcat gccagttccc gtgcttgaag ccggccgccc
      481 gcagcatgcc gcggggggca tatccgagcg cctcgtgcat gcgcacgctc gggtcgttgg
      541 gcagcccgat gacagcgacc acgctcttga agccctgtgc ctccagggac ttcagcaggt
      601 gggtgtagag cgtggagccc agtcccgtcc gctggtggcg gggggacacg tacacggtcg
      661 actcggccgt ccagtcgtag gcgttgcgtg ccttccaggg acccgcgtag gcgatgccgg
      721 cgacctcgcc gtccacctcg gcgacgagcc agggatagcg ctcccgcaga cggacgaggt
      781 cgtccgtcca ctcctgcggt tcctgcggct cggtacggaa gttgaccgtg cttgtctcga
      841 tgtagtggtt gacgatggtg cagaccgccg gcatgtccgc ctcggtggca cggcggatgt
      901 cggccgggcg tcgttctggg ctcatggtag atcccctcga tcgagttgag agtgaatatg
      961 agactctaat tggataccga ggggaattta tggaacgtca gtggagcatt tttgacaaga
     1021 aatatttgct agctgatagt gaccttaggc gacttttgaa cgcgcaataa tggtttctga
     1081 cgtatgtgct tagctcatta aactccagaa acccgcggct cagtggctcc ttcaacgttg
     1141 cggttctgtc agttccaaac gtaaaacggc ttgtcccgcg tcatcggcgg gggtcataac
     1201 gtgactccct taattctccg ctcatgatcg ataacatgtc gactatcgga cgagctcaaa
     1261 gtttaacgtg tgaagacgtg cttctaatgt agtttaaact gaaggcggga aacgacaatc
     1321 tgatccaagc tcaagctaag cttgcatgcc tgcaggatat cgtggatcca agcttgccac
     1381 gtgccgccac gtgccgccac gtgccgccac gtgcctctag aggatccatc tccactgacg
     1441 taagggatga cgcacaatcc cactatcctt cgcaagaccc ttcctctata taaggaagtt
     1501 catttcattt ggagaggaca cgctgggatc cccaccatga gctacggcga cagcagagag
     1561 aagatcctga gcgccgccac ccggctgttc cagctgcaag gctactacgg caccggcctg
     1621 aaccagatca tcaaagagag cggagccccc aagggcagcc tgtactacca cttccctggc
     1681 ggcaaagagc agctggccat cgaggccgtg aacgagatga aggagtacat ccggcagaaa
     1741 atcgccgact gcatggaagc ctgcaccgac cccgccgagg gcatccaggc ctttctgaaa
     1801 gagctgagct gccagttctc ctgcaccgag gacatcgagg gcctgcctgt gggactgctg
     1861 gccgccgaga caagcctgaa gtccgagccc ctgagagagg cctgccacga ggcctacaaa
     1921 gaatgggcca gcgtgtacga ggaaaagctg cggcagaccg gctgcagcga gagcagagcc
     1981 aaagaggcca gcaccgtcgt caacgccatg atcgagggcg gcatcctgct gagcctgacc
     2041 gccaagaaca gcacccccct gctgcacatc agcagctgca tccccgacct gctgaagaga
     2101 gcccccccga ccgacgtcag cctgggggac gagctccact tagacggcga ggacgtggcg
     2161 atggcgcatg ccgacgcgct agacgatttc gatctggaca tgttggggga cggggattcc
     2221 ccggggccgg gatttacccc ccacgactcc gccccctacg gcgctctgga tacggccgac
     2281 ttcgagtttg agcagatgtt taccgatgcc cttggaattg acgagtacgg tgggccgaag
     2341 aagaagagga aggtttagtg tcggtacgct gaaatcacca gtctctctct acaaatctat
     2401 ctctctctat tttctccata aataatgtgt gagtagtttc ccgataaggg aaattagggt
     2461 tcttataggg tttcgctcat gtgttgagca tataagaaac ccttagtatg tatttgtatt
     2521 tgtaaaatac ttctatcaat aaaatttcta attcctaaaa ccaaaatcca gtactaaaat
     2581 ccagatcgat gcttgatact acgtcttcgg gctagagcgg actagttcta gagcggccgc
     2641 caccgcggtg gagctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt
     2701 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     2761 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     2821 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     2881 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     2941 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3001 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgaaggcct
     3061 tgacaggata tattggcggg taaactaagt cgctgtatgt gtttgtttga gatctcatgt
     3121 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3181 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3241 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3301 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3361 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3421 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3481 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3541 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3601 acggctacac tagaagaaca gtatttggta tctgtgctct gctgaagcca gttaccttcg
     3661 gaagaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3721 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3781 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3841 attcttgtta gacattattt gccgactacc ttggtgatct cgcctttcac gtagtgaaca
     3901 aattcttcca actgatctgc gcgcgaggcc aagcgatctt cttgtccaag ataagcctgc
     3961 ctagcttcaa gtatgacggg ctgatactgg gccggcaggc gctccattgc ccagtcggca
     4021 gcgacatcct tcggcgcgat tttgccggtt actgcgctgt accaaatgcg ggacaacgta
     4081 agcactacat ttcgctcatc gccagcccag tcgggcggcg agttccatag cgttaaggtt
     4141 tcatttagcg cctcaaatag atcctgttca ggaaccggat caaagagttc ctccgccgct
     4201 ggacctacca aggcaacgct atgttctctt gcttttgtca gcaagatagc cagatcaatg
     4261 tcgatcgtgg ctggctcgaa gatacctgca agaatgtcat tgcgctgcca ttctccaaat
     4321 tgcagttcgc gcttagctgg ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg
     4381 acttctacag cgcggagaat ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg
     4441 ttgatcaaag ctcgccgcgt tgtttcatca agccttacgg tcaccgtaac cagcaaatca
     4501 atatcactgt gtggcttcag gccgccatcc actgcggagc cgtacaaatg tacggccagc
     4561 aacgtcggtt cgagatggcg ctcgatgacg ccaactacct ctgatagttg agtcgatact
     4621 tcggcgatca ccgcttccct catgatgttt aactcctgaa ttaagccgcg ccgcgaagcg
     4681 gtgtcggctt gaatgaattg ttaggcgtca tcctgtgctc ccgagaacca gtaccagtac
     4741 atcgctgttt cgttcgagac ttgaggtcta gttttatacg tgaacaggtc aatgccgccg
     4801 agagtaaagc cacattttgc gtacaaattg caggcaggta cattgttcgt ttgtgccatg
     4861 atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacaca acgtggcttt
     4921 gttgaataaa tcgaactttt gctgagttga aggatcagat cacgcatctt cccgacaacg
     4981 cagaccgttc cgtggcaaag caaaagttca aaatcaccaa ctggtccacc tacaacaaag
     5041 ctctcatcaa ccgtggctcc ctcactttct ggctggatga tggggcgatt caggagatcc
     5101 ccatccaaca gcccgccgtc gagcgggctt ttttatcccc ggaagcctgt ggatagaggg
     5161 tagttatcca cgtgaaaccg ctaatgcccc gcaaagcctt gattcacggg gctttccggc
     5221 ccgctccaaa aactatccac gtgaaatcgc taatcagggt acgtgaaatc gctaatcgga
     5281 gtacgtgaaa tcgctaataa ggtcacgtga aatcgctaat caaaaaggca cgtgagaacg
     5341 ctaatagccc tttcagatca acagcttgca aacacccctc gctccggcaa gtagttacag
     5401 caagtagtat gttcaattag cttttcaatt atgaatatat atatcaatta ttggtcgccc
     5461 ttggcttgtg gacaatgcgc tacgcgcacc ggctccgccc gtggacaacc gcaagcggtt
     5521 gcccaccgtc gagcgccttt gcccacaacc cggcggccgg ccgcaacaga tcgttttata
     5581 aatttttttt tttgaaaaag aaaaagcccg aaaggcggca acctctcggg cttctggatt
     5641 tccgatcccc ggaattagac tctggctgcc tgtatcgagt ggtgattttg tgccgagctg
     5701 ccggtcgggg agctgttggc
//
