LOCUS       sequence-387029-        5720 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proG1090::QacR-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: JAB2165
FEATURES             Location/Qualifiers
     source          1..5720
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    21..45
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(115..367)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(387..938)
                     /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(980..1159)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     1476..1496
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        1485..1531
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             2114..2347
                     /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             2348..2368
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2424..2600
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(2682..2702)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2682..2700)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2721..2737)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2721..2737)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2734..2756)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2745..2761
                     /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(2769..2775,2776..2793,2794..2799))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2814..2835
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2952..2969)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3074..3098
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(3189..3777)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3269..3288)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3868..4656)
                     /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      5114..5549
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 gggagctgtt ggctggctgg tggcaggata tattgtggtg taaacaaatt gacgcttaga
       61 caacttaata acacattgcg gacgttttta atgtaatcag cttgcatgcc ggtcgatcta
      121 gtaacataga tgacaccgcg cgcgataatt tatcctagtt tgcgcgctat attttgtttt
      181 ctatcgcgta ttaaatgtat aattgcggga ctctaatcat aaaaacccat ctcataaata
      241 acgtcatgca ttacatgtta attattacat gcttaacgta attcaacaga aattatatga
      301 taatcatcgc aagaccggca acaggattca atcttaagaa actttattgc caaatgtttg
      361 aacgatctgc ttgactctag gggtcatcag atttcggtga cgggcaggac cggacggggc
      421 ggcaccggca ggctgaagtc cagctgccag aaacccacgt catgccagtt cccgtgcttg
      481 aagccggccg cccgcagcat gccgcggggg gcatatccga gcgcctcgtg catgcgcacg
      541 ctcgggtcgt tgggcagccc gatgacagcg accacgctct tgaagccctg tgcctccagg
      601 gacttcagca ggtgggtgta gagcgtggag cccagtcccg tccgctggtg gcggggggac
      661 acgtacacgg tcgactcggc cgtccagtcg taggcgttgc gtgccttcca gggacccgcg
      721 taggcgatgc cggcgacctc gccgtccacc tcggcgacga gccagggata gcgctcccgc
      781 agacggacga ggtcgtccgt ccactcctgc ggttcctgcg gctcggtacg gaagttgacc
      841 gtgcttgtct cgatgtagtg gttgacgatg gtgcagaccg ccggcatgtc cgcctcggtg
      901 gcacggcgga tgtcggccgg gcgtcgttct gggctcatgg tagatcccct cgatcgagtt
      961 gagagtgaat atgagactct aattggatac cgaggggaat ttatggaacg tcagtggagc
     1021 atttttgaca agaaatattt gctagctgat agtgacctta ggcgactttt gaacgcgcaa
     1081 taatggtttc tgacgtatgt gcttagctca ttaaactcca gaaacccgcg gctcagtggc
     1141 tccttcaacg ttgcggttct gtcagttcca aacgtaaaac ggcttgtccc gcgtcatcgg
     1201 cgggggtcat aacgtgactc ccttaattct ccgctcatga tcgataacat gtcgactatc
     1261 ggacgagctc aaagtttaac gtgtgaagac gtgcttctaa tgtagtttaa actgaaggcg
     1321 ggaaacgaca atctgatcca agctcaagct aagcttgcat gcctgcagga tatcgtggat
     1381 ccaagcttgc cacgtgccgc cacgtgccgc cacgtgccgc cacgtgcctc tagaggatcc
     1441 atctccactg acgtaaggga tgacgcacaa tcccactatc cttcgcaaga cccttcctct
     1501 atataaggaa gttcatttca tttggagagg acacgctggg atccccacca tgaacctgaa
     1561 ggacaagatc ctgggcgtgg ccaaagagct gttcatcaag aacggctaca acgccaccac
     1621 caccggcgag atcgtgaagc tgagcgagag cagcaagggc aacctgtact accacttcaa
     1681 gaccaaagag aacctgttcc tcgagatcct gaacatcgag gaatccaagt ggcaggaaca
     1741 gtggaaaaaa gaacagatca agtgcaagac aaaccgcgag aagttctacc tgtacaacga
     1801 gctgagcctg accaccgagt actactaccc cctgcagaac gccatcatcg agttctacac
     1861 agagtactat aagaccaaca gcatcaacga gaagatgaac aagctcgaga acaagtacat
     1921 cgacgcctac cacgtgatct tcaaagaggg caatctgaac ggcgagtggt gcatcaatga
     1981 cgtgaacgcc gtgtccaaga tcgccgccaa tgccgtgaat ggcatcgtga ccttcaccca
     2041 cgagcagaac attaacgagc ggatcaagct catgaacaaa ttcagccaga tcttcctgaa
     2101 cgggctgagc aaggcccccc cgaccgacgt cagcctgggg gacgagctcc acttagacgg
     2161 cgaggacgtg gcgatggcgc atgccgacgc gctagacgat ttcgatctgg acatgttggg
     2221 ggacggggat tccccggggc cgggatttac cccccacgac tccgccccct acggcgctct
     2281 ggatacggcc gacttcgagt ttgagcagat gtttaccgat gcccttggaa ttgacgagta
     2341 cggtgggccg aagaagaaga ggaaggttta gtgtcggtac gctgaaatca ccagtctctc
     2401 tctacaaatc tatctctctc tattttctcc ataaataatg tgtgagtagt ttcccgataa
     2461 gggaaattag ggttcttata gggtttcgct catgtgttga gcatataaga aacccttagt
     2521 atgtatttgt atttgtaaaa tacttctatc aataaaattt ctaattccta aaaccaaaat
     2581 ccagtactaa aatccagatc gatgcttgat actacgtctt cgggctagag cggactagtt
     2641 ctagagcggc cgccaccgcg gtggagctcc agcttttgtt ccctttagtg agggttaatt
     2701 gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     2761 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     2821 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     2881 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     2941 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3001 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3061 aacatgaagg ccttgacagg atatattggc gggtaaacta agtcgctgta tgtgtttgtt
     3121 tgagatctca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3181 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3241 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3301 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3361 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3421 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3481 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3541 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3601 tggtggccta actacggcta cactagaaga acagtatttg gtatctgtgc tctgctgaag
     3661 ccagttacct tcggaagaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3721 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3781 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3841 attttggtca tgaattcttg ttagacatta tttgccgact accttggtga tctcgccttt
     3901 cacgtagtga acaaattctt ccaactgatc tgcgcgcgag gccaagcgat cttcttgtcc
     3961 aagataagcc tgcctagctt caagtatgac gggctgatac tgggccggca ggcgctccat
     4021 tgcccagtcg gcagcgacat ccttcggcgc gattttgccg gttactgcgc tgtaccaaat
     4081 gcgggacaac gtaagcacta catttcgctc atcgccagcc cagtcgggcg gcgagttcca
     4141 tagcgttaag gtttcattta gcgcctcaaa tagatcctgt tcaggaaccg gatcaaagag
     4201 ttcctccgcc gctggaccta ccaaggcaac gctatgttct cttgcttttg tcagcaagat
     4261 agccagatca atgtcgatcg tggctggctc gaagatacct gcaagaatgt cattgcgctg
     4321 ccattctcca aattgcagtt cgcgcttagc tggataacgc cacggaatga tgtcgtcgtg
     4381 cacaacaatg gtgacttcta cagcgcggag aatctcgctc tctccagggg aagccgaagt
     4441 ttccaaaagg tcgttgatca aagctcgccg cgttgtttca tcaagcctta cggtcaccgt
     4501 aaccagcaaa tcaatatcac tgtgtggctt caggccgcca tccactgcgg agccgtacaa
     4561 atgtacggcc agcaacgtcg gttcgagatg gcgctcgatg acgccaacta cctctgatag
     4621 ttgagtcgat acttcggcga tcaccgcttc cctcatgatg tttaactcct gaattaagcc
     4681 gcgccgcgaa gcggtgtcgg cttgaatgaa ttgttaggcg tcatcctgtg ctcccgagaa
     4741 ccagtaccag tacatcgctg tttcgttcga gacttgaggt ctagttttat acgtgaacag
     4801 gtcaatgccg ccgagagtaa agccacattt tgcgtacaaa ttgcaggcag gtacattgtt
     4861 cgtttgtgcc atgatgatat atttttatct tgtgcaatgt aacatcagag attttgagac
     4921 acaacgtggc tttgttgaat aaatcgaact tttgctgagt tgaaggatca gatcacgcat
     4981 cttcccgaca acgcagaccg ttccgtggca aagcaaaagt tcaaaatcac caactggtcc
     5041 acctacaaca aagctctcat caaccgtggc tccctcactt tctggctgga tgatggggcg
     5101 attcaggaga tccccatcca acagcccgcc gtcgagcggg cttttttatc cccggaagcc
     5161 tgtggataga gggtagttat ccacgtgaaa ccgctaatgc cccgcaaagc cttgattcac
     5221 ggggctttcc ggcccgctcc aaaaactatc cacgtgaaat cgctaatcag ggtacgtgaa
     5281 atcgctaatc ggagtacgtg aaatcgctaa taaggtcacg tgaaatcgct aatcaaaaag
     5341 gcacgtgaga acgctaatag ccctttcaga tcaacagctt gcaaacaccc ctcgctccgg
     5401 caagtagtta cagcaagtag tatgttcaat tagcttttca attatgaata tatatatcaa
     5461 ttattggtcg cccttggctt gtggacaatg cgctacgcgc accggctccg cccgtggaca
     5521 accgcaagcg gttgcccacc gtcgagcgcc tttgcccaca acccggcggc cggccgcaac
     5581 agatcgtttt ataaattttt ttttttgaaa aagaaaaagc ccgaaaggcg gcaacctctc
     5641 gggcttctgg atttccgatc cccggaatta gactctggct gcctgtatcg agtggtgatt
     5701 ttgtgccgag ctgccggtcg
//
