LOCUS       sequence-387094-        5750 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proG1090::BetI-VP16_19St).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5750)
  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 5750)
  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: JAB2495
FEATURES             Location/Qualifiers
     source          1..5750
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(86..338)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(358..909)
                     /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(951..1130)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     1447..1467
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        1456..1502
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             2115..2348
                     /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             2349..2369
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2425..2601
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(2683..2703)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2683..2701)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2722..2738)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2722..2738)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2735..2757)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2746..2762
                     /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(2770..2776,2777..2794,2795..2800))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2815..2836
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2953..2970)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3075..3099
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(3190..3778)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3270..3289)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3869..4657)
                     /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      5115..5550
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(5742..5750,1..16)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 atattgtggt gtaaacaaat tgacgcttag acaacttaat aacacattgc ggacgttttt
       61 aatgtaatca gcttgcatgc cggtcgatct agtaacatag atgacaccgc gcgcgataat
      121 ttatcctagt ttgcgcgcta tattttgttt tctatcgcgt attaaatgta taattgcggg
      181 actctaatca taaaaaccca tctcataaat aacgtcatgc attacatgtt aattattaca
      241 tgcttaacgt aattcaacag aaattatatg ataatcatcg caagaccggc aacaggattc
      301 aatcttaaga aactttattg ccaaatgttt gaacgatctg cttgactcta ggggtcatca
      361 gatttcggtg acgggcagga ccggacgggg cggcaccggc aggctgaagt ccagctgcca
      421 gaaacccacg tcatgccagt tcccgtgctt gaagccggcc gcccgcagca tgccgcgggg
      481 ggcatatccg agcgcctcgt gcatgcgcac gctcgggtcg ttgggcagcc cgatgacagc
      541 gaccacgctc ttgaagccct gtgcctccag ggacttcagc aggtgggtgt agagcgtgga
      601 gcccagtccc gtccgctggt ggcgggggga cacgtacacg gtcgactcgg ccgtccagtc
      661 gtaggcgttg cgtgccttcc agggacccgc gtaggcgatg ccggcgacct cgccgtccac
      721 ctcggcgacg agccagggat agcgctcccg cagacggacg aggtcgtccg tccactcctg
      781 cggttcctgc ggctcggtac ggaagttgac cgtgcttgtc tcgatgtagt ggttgacgat
      841 ggtgcagacc gccggcatgt ccgcctcggt ggcacggcgg atgtcggccg ggcgtcgttc
      901 tgggctcatg gtagatcccc tcgatcgagt tgagagtgaa tatgagactc taattggata
      961 ccgaggggaa tttatggaac gtcagtggag catttttgac aagaaatatt tgctagctga
     1021 tagtgacctt aggcgacttt tgaacgcgca ataatggttt ctgacgtatg tgcttagctc
     1081 attaaactcc agaaacccgc ggctcagtgg ctccttcaac gttgcggttc tgtcagttcc
     1141 aaacgtaaaa cggcttgtcc cgcgtcatcg gcgggggtca taacgtgact cccttaattc
     1201 tccgctcatg atcgataaca tgtcgactat cggacgagct caaagtttaa cgtgtgaaga
     1261 cgtgcttcta atgtagttta aactgaaggc gggaaacgac aatctgatcc aagctcaagc
     1321 taagcttgca tgcctgcagg atatcgtgga tccaagcttg ccacgtgccg ccacgtgccg
     1381 ccacgtgccg ccacgtgcct ctagaggatc catctccact gacgtaaggg atgacgcaca
     1441 atcccactat ccttcgcaag acccttcctc tatataagga agttcatttc atttggagag
     1501 gacacgctgg gatccccacc atgccgaaac tgggtatgca gagcattcgt cgtcgtcagc
     1561 tgattgatgc aaccctggaa gcaattaatg aagttggtat gcatgatgca accattgcac
     1621 agattgcacg tcgtgccggt gttagcaccg gtattattag ccattatttc cgcgataaaa
     1681 acggtctgct ggaagcaacc atgcgtgata ttaccagcca gctgcgtgat gcagttctga
     1741 atcgtctgca tgcactgccg cagggtagcg cagaacagcg tctgcaggca attgttggtg
     1801 gtaattttga tgaaacccag gttagcagcg cagcaatgaa agcatggctg gcattttggg
     1861 caagcagcat gcatcagccg atgctgtatc gtctgcagca ggttagcagt cgtcgtctgc
     1921 tgagcaatct ggttagcgaa tttcgtcgtg aactgcctcg tgaacaggca caagaggcag
     1981 gttatggtct ggcagcactg attgatggtc tgtggctgcg tgcagcactg agcggtaaac
     2041 cgctggataa aacccgtgca aatagcctga cccgtcattt tatcacccag catctgccga
     2101 ccgatggagg aggtgccccc ccgaccgacg tcagcctggg ggacgagctc cacttagacg
     2161 gcgaggacgt ggcgatggcg catgccgacg cgctagacga tttcgatctg gacatgttgg
     2221 gggacgggga ttccccgggg ccgggattta ccccccacga ctccgccccc tacggcgctc
     2281 tggatacggc cgacttcgag tttgagcaga tgtttaccga tgcccttgga attgacgagt
     2341 acggtgggcc gaagaagaag aggaaggttt agtgtcggta cgctgaaatc accagtctct
     2401 ctctacaaat ctatctctct ctattttctc cataaataat gtgtgagtag tttcccgata
     2461 agggaaatta gggttcttat agggtttcgc tcatgtgttg agcatataag aaacccttag
     2521 tatgtatttg tatttgtaaa atacttctat caataaaatt tctaattcct aaaaccaaaa
     2581 tccagtacta aaatccagat cgatgcttga tactacgtct tcgggctaga gcggactagt
     2641 tctagagcgg ccgccaccgc ggtggagctc cagcttttgt tccctttagt gagggttaat
     2701 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     2761 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     2821 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     2881 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     2941 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3001 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3061 gaacatgaag gccttgacag gatatattgg cgggtaaact aagtcgctgt atgtgtttgt
     3121 ttgagatctc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3181 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3241 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3301 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3361 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3421 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3481 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3541 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3601 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgtg ctctgctgaa
     3661 gccagttacc ttcggaagaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3721 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3781 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3841 gattttggtc atgaattctt gttagacatt atttgccgac taccttggtg atctcgcctt
     3901 tcacgtagtg aacaaattct tccaactgat ctgcgcgcga ggccaagcga tcttcttgtc
     3961 caagataagc ctgcctagct tcaagtatga cgggctgata ctgggccggc aggcgctcca
     4021 ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc ggttactgcg ctgtaccaaa
     4081 tgcgggacaa cgtaagcact acatttcgct catcgccagc ccagtcgggc ggcgagttcc
     4141 atagcgttaa ggtttcattt agcgcctcaa atagatcctg ttcaggaacc ggatcaaaga
     4201 gttcctccgc cgctggacct accaaggcaa cgctatgttc tcttgctttt gtcagcaaga
     4261 tagccagatc aatgtcgatc gtggctggct cgaagatacc tgcaagaatg tcattgcgct
     4321 gccattctcc aaattgcagt tcgcgcttag ctggataacg ccacggaatg atgtcgtcgt
     4381 gcacaacaat ggtgacttct acagcgcgga gaatctcgct ctctccaggg gaagccgaag
     4441 tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc atcaagcctt acggtcaccg
     4501 taaccagcaa atcaatatca ctgtgtggct tcaggccgcc atccactgcg gagccgtaca
     4561 aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat gacgccaact acctctgata
     4621 gttgagtcga tacttcggcg atcaccgctt ccctcatgat gtttaactcc tgaattaagc
     4681 cgcgccgcga agcggtgtcg gcttgaatga attgttaggc gtcatcctgt gctcccgaga
     4741 accagtacca gtacatcgct gtttcgttcg agacttgagg tctagtttta tacgtgaaca
     4801 ggtcaatgcc gccgagagta aagccacatt ttgcgtacaa attgcaggca ggtacattgt
     4861 tcgtttgtgc catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga
     4921 cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc agatcacgca
     4981 tcttcccgac aacgcagacc gttccgtggc aaagcaaaag ttcaaaatca ccaactggtc
     5041 cacctacaac aaagctctca tcaaccgtgg ctccctcact ttctggctgg atgatggggc
     5101 gattcaggag atccccatcc aacagcccgc cgtcgagcgg gcttttttat ccccggaagc
     5161 ctgtggatag agggtagtta tccacgtgaa accgctaatg ccccgcaaag ccttgattca
     5221 cggggctttc cggcccgctc caaaaactat ccacgtgaaa tcgctaatca gggtacgtga
     5281 aatcgctaat cggagtacgt gaaatcgcta ataaggtcac gtgaaatcgc taatcaaaaa
     5341 ggcacgtgag aacgctaata gccctttcag atcaacagct tgcaaacacc cctcgctccg
     5401 gcaagtagtt acagcaagta gtatgttcaa ttagcttttc aattatgaat atatatatca
     5461 attattggtc gcccttggct tgtggacaat gcgctacgcg caccggctcc gcccgtggac
     5521 aaccgcaagc ggttgcccac cgtcgagcgc ctttgcccac aacccggcgg ccggccgcaa
     5581 cagatcgttt tataaatttt tttttttgaa aaagaaaaag cccgaaaggc ggcaacctct
     5641 cgggcttctg gatttccgat ccccggaatt agactctggc tgcctgtatc gagtggtgat
     5701 tttgtgccga gctgccggtc ggggagctgt tggctggctg gtggcaggat
//
