LOCUS       sequence-393421-        5477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proG1090::IcaR-NLS_19St).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5477)
  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 5477)
  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: JAB2488
FEATURES             Location/Qualifiers
     source          1..5477
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    12..36
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(106..358)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(378..929)
                     /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(971..1150)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     1467..1487
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        1476..1522
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             2096..2116
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2172..2348
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(2430..2450)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2430..2448)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2469..2485)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2469..2485)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2482..2504)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2493..2509
                     /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(2517..2523,2524..2541,2542..2547))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2562..2583
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2700..2717)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2822..2846
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(2937..3525)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3017..3036)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3616..4404)
                     /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      4862..5297
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 tggctggctg gtggcaggat atattgtggt gtaaacaaat tgacgcttag acaacttaat
       61 aacacattgc ggacgttttt aatgtaatca gcttgcatgc cggtcgatct agtaacatag
      121 atgacaccgc gcgcgataat ttatcctagt ttgcgcgcta tattttgttt tctatcgcgt
      181 attaaatgta taattgcggg actctaatca taaaaaccca tctcataaat aacgtcatgc
      241 attacatgtt aattattaca tgcttaacgt aattcaacag aaattatatg ataatcatcg
      301 caagaccggc aacaggattc aatcttaaga aactttattg ccaaatgttt gaacgatctg
      361 cttgactcta ggggtcatca gatttcggtg acgggcagga ccggacgggg cggcaccggc
      421 aggctgaagt ccagctgcca gaaacccacg tcatgccagt tcccgtgctt gaagccggcc
      481 gcccgcagca tgccgcgggg ggcatatccg agcgcctcgt gcatgcgcac gctcgggtcg
      541 ttgggcagcc cgatgacagc gaccacgctc ttgaagccct gtgcctccag ggacttcagc
      601 aggtgggtgt agagcgtgga gcccagtccc gtccgctggt ggcgggggga cacgtacacg
      661 gtcgactcgg ccgtccagtc gtaggcgttg cgtgccttcc agggacccgc gtaggcgatg
      721 ccggcgacct cgccgtccac ctcggcgacg agccagggat agcgctcccg cagacggacg
      781 aggtcgtccg tccactcctg cggttcctgc ggctcggtac ggaagttgac cgtgcttgtc
      841 tcgatgtagt ggttgacgat ggtgcagacc gccggcatgt ccgcctcggt ggcacggcgg
      901 atgtcggccg ggcgtcgttc tgggctcatg gtagatcccc tcgatcgagt tgagagtgaa
      961 tatgagactc taattggata ccgaggggaa tttatggaac gtcagtggag catttttgac
     1021 aagaaatatt tgctagctga tagtgacctt aggcgacttt tgaacgcgca ataatggttt
     1081 ctgacgtatg tgcttagctc attaaactcc agaaacccgc ggctcagtgg ctccttcaac
     1141 gttgcggttc tgtcagttcc aaacgtaaaa cggcttgtcc cgcgtcatcg gcgggggtca
     1201 taacgtgact cccttaattc tccgctcatg atcgataaca tgtcgactat cggacgagct
     1261 caaagtttaa cgtgtgaaga cgtgcttcta atgtagttta aactgaaggc gggaaacgac
     1321 aatctgatcc aagctcaagc taagcttgca tgcctgcagg atatcgtgga tccaagcttg
     1381 ccacgtgccg ccacgtgccg ccacgtgccg ccacgtgcct ctagaggatc catctccact
     1441 gacgtaaggg atgacgcaca atcccactat ccttcgcaag acccttcctc tatataagga
     1501 agttcatttc atttggagag gacacgctgg gatccccacc atgaaggaca agatcatcga
     1561 caacgccatc accctgttca gcgagaaggg ctacgacggc accacactgg acgacatcag
     1621 caagagcgtg aacatcaaga aggccagcct gtactaccac tacgacaaca aagaggaaat
     1681 ctaccggaag tccgtcgaga actgcttcaa ctacttcatc gactttctgc tgcggaacca
     1741 cgacgataac tacagcatcg acggcctgta ccagttcctg ttcaagttta tcttcgacgt
     1801 ggacgagcgg tacatcaagc tgtacgtgca gctgagcagc gcccctgagg ccctgaacag
     1861 cgagatcaag caccatctgc aggaaatcaa caccaccctg cacgacgagc tgatcaagta
     1921 ctacgacccc acccacattg ctctgaacaa agaagatttc atcaacctga tcctgctgtt
     1981 cctggaaacc tggtacttcc gggccagctt ctcccagaaa ttcggcgtga tcgaggacag
     2041 caagaaccgg ttcaaggacc aggtgtacag cctgctgaac gtgttcctga agaagccgaa
     2101 gaagaagagg aaggtttagt gtcggtacgc tgaaatcacc agtctctctc tacaaatcta
     2161 tctctctcta ttttctccat aaataatgtg tgagtagttt cccgataagg gaaattaggg
     2221 ttcttatagg gtttcgctca tgtgttgagc atataagaaa cccttagtat gtatttgtat
     2281 ttgtaaaata cttctatcaa taaaatttct aattcctaaa accaaaatcc agtactaaaa
     2341 tccagatcga tgcttgatac tacgtcttcg ggctagagcg gactagttct agagcggccg
     2401 ccaccgcggt ggagctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
     2461 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     2521 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     2581 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     2641 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     2701 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     2761 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgaaggcc
     2821 ttgacaggat atattggcgg gtaaactaag tcgctgtatg tgtttgtttg agatctcatg
     2881 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2941 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3001 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3061 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3121 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3181 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3241 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3301 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3361 tacggctaca ctagaagaac agtatttggt atctgtgctc tgctgaagcc agttaccttc
     3421 ggaagaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3481 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3541 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3601 aattcttgtt agacattatt tgccgactac cttggtgatc tcgcctttca cgtagtgaac
     3661 aaattcttcc aactgatctg cgcgcgaggc caagcgatct tcttgtccaa gataagcctg
     3721 cctagcttca agtatgacgg gctgatactg ggccggcagg cgctccattg cccagtcggc
     3781 agcgacatcc ttcggcgcga ttttgccggt tactgcgctg taccaaatgc gggacaacgt
     3841 aagcactaca tttcgctcat cgccagccca gtcgggcggc gagttccata gcgttaaggt
     3901 ttcatttagc gcctcaaata gatcctgttc aggaaccgga tcaaagagtt cctccgccgc
     3961 tggacctacc aaggcaacgc tatgttctct tgcttttgtc agcaagatag ccagatcaat
     4021 gtcgatcgtg gctggctcga agatacctgc aagaatgtca ttgcgctgcc attctccaaa
     4081 ttgcagttcg cgcttagctg gataacgcca cggaatgatg tcgtcgtgca caacaatggt
     4141 gacttctaca gcgcggagaa tctcgctctc tccaggggaa gccgaagttt ccaaaaggtc
     4201 gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa ccagcaaatc
     4261 aatatcactg tgtggcttca ggccgccatc cactgcggag ccgtacaaat gtacggccag
     4321 caacgtcggt tcgagatggc gctcgatgac gccaactacc tctgatagtt gagtcgatac
     4381 ttcggcgatc accgcttccc tcatgatgtt taactcctga attaagccgc gccgcgaagc
     4441 ggtgtcggct tgaatgaatt gttaggcgtc atcctgtgct cccgagaacc agtaccagta
     4501 catcgctgtt tcgttcgaga cttgaggtct agttttatac gtgaacaggt caatgccgcc
     4561 gagagtaaag ccacattttg cgtacaaatt gcaggcaggt acattgttcg tttgtgccat
     4621 gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtggctt
     4681 tgttgaataa atcgaacttt tgctgagttg aaggatcaga tcacgcatct tcccgacaac
     4741 gcagaccgtt ccgtggcaaa gcaaaagttc aaaatcacca actggtccac ctacaacaaa
     4801 gctctcatca accgtggctc cctcactttc tggctggatg atggggcgat tcaggagatc
     4861 cccatccaac agcccgccgt cgagcgggct tttttatccc cggaagcctg tggatagagg
     4921 gtagttatcc acgtgaaacc gctaatgccc cgcaaagcct tgattcacgg ggctttccgg
     4981 cccgctccaa aaactatcca cgtgaaatcg ctaatcaggg tacgtgaaat cgctaatcgg
     5041 agtacgtgaa atcgctaata aggtcacgtg aaatcgctaa tcaaaaaggc acgtgagaac
     5101 gctaatagcc ctttcagatc aacagcttgc aaacacccct cgctccggca agtagttaca
     5161 gcaagtagta tgttcaatta gcttttcaat tatgaatata tatatcaatt attggtcgcc
     5221 cttggcttgt ggacaatgcg ctacgcgcac cggctccgcc cgtggacaac cgcaagcggt
     5281 tgcccaccgt cgagcgcctt tgcccacaac ccggcggccg gccgcaacag atcgttttat
     5341 aaattttttt ttttgaaaaa gaaaaagccc gaaaggcggc aacctctcgg gcttctggat
     5401 ttccgatccc cggaattaga ctctggctgc ctgtatcgag tggtgatttt gtgccgagct
     5461 gccggtcggg gagctgt
//
