LOCUS       sequence-387079-        5717 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proG1090::AmtR-ERF4(RD)-19St).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5717)
  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 5717)
  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: JAB2282
FEATURES             Location/Qualifiers
     source          1..5717
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    6..30
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(100..352)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(372..923)
                     /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(965..1144)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     1461..1481
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        1470..1516
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             2330..2350
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2406..2582
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(2664..2684)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2664..2682)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2703..2719)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2703..2719)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2716..2738)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2727..2743
                     /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(2751..2757,2758..2775,2776..2781))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2796..2817
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2934..2951)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3056..3080
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(3171..3759)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3251..3270)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3850..4638)
                     /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      5096..5531
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 gctggtggca ggatatattg tggtgtaaac aaattgacgc ttagacaact taataacaca
       61 ttgcggacgt ttttaatgta atcagcttgc atgccggtcg atctagtaac atagatgaca
      121 ccgcgcgcga taatttatcc tagtttgcgc gctatatttt gttttctatc gcgtattaaa
      181 tgtataattg cgggactcta atcataaaaa cccatctcat aaataacgtc atgcattaca
      241 tgttaattat tacatgctta acgtaattca acagaaatta tatgataatc atcgcaagac
      301 cggcaacagg attcaatctt aagaaacttt attgccaaat gtttgaacga tctgcttgac
      361 tctaggggtc atcagatttc ggtgacgggc aggaccggac ggggcggcac cggcaggctg
      421 aagtccagct gccagaaacc cacgtcatgc cagttcccgt gcttgaagcc ggccgcccgc
      481 agcatgccgc ggggggcata tccgagcgcc tcgtgcatgc gcacgctcgg gtcgttgggc
      541 agcccgatga cagcgaccac gctcttgaag ccctgtgcct ccagggactt cagcaggtgg
      601 gtgtagagcg tggagcccag tcccgtccgc tggtggcggg gggacacgta cacggtcgac
      661 tcggccgtcc agtcgtaggc gttgcgtgcc ttccagggac ccgcgtaggc gatgccggcg
      721 acctcgccgt ccacctcggc gacgagccag ggatagcgct cccgcagacg gacgaggtcg
      781 tccgtccact cctgcggttc ctgcggctcg gtacggaagt tgaccgtgct tgtctcgatg
      841 tagtggttga cgatggtgca gaccgccggc atgtccgcct cggtggcacg gcggatgtcg
      901 gccgggcgtc gttctgggct catggtagat cccctcgatc gagttgagag tgaatatgag
      961 actctaattg gataccgagg ggaatttatg gaacgtcagt ggagcatttt tgacaagaaa
     1021 tatttgctag ctgatagtga ccttaggcga cttttgaacg cgcaataatg gtttctgacg
     1081 tatgtgctta gctcattaaa ctccagaaac ccgcggctca gtggctcctt caacgttgcg
     1141 gttctgtcag ttccaaacgt aaaacggctt gtcccgcgtc atcggcgggg gtcataacgt
     1201 gactccctta attctccgct catgatcgat aacatgtcga ctatcggacg agctcaaagt
     1261 ttaacgtgtg aagacgtgct tctaatgtag tttaaactga aggcgggaaa cgacaatctg
     1321 atccaagctc aagctaagct tgcatgcctg caggatatcg tggatccaag cttgccacgt
     1381 gccgccacgt gccgccacgt gccgccacgt gcctctagag gatccatctc cactgacgta
     1441 agggatgacg cacaatccca ctatccttcg caagaccctt cctctatata aggaagttca
     1501 tttcatttgg agaggacacg ctgggatccc caccatggct ggcgccgtgg gcagacccag
     1561 aagatctgct cctcggagag ccggcaagaa cccccgggaa gagattctgg atgccagcgc
     1621 cgagctgttc accagacagg gctttgccac caccagcacc caccagattg ccgacgctgt
     1681 gggcatcaga caggccagcc tgtactacca cttccccagc aagaccgaga tcttcctgac
     1741 cctgctgaaa agcaccgtgg aaccctccac cgtgctggcc gaggatctgt ctaccctgga
     1801 cgccggaccc gaaatgagac tgtgggctat cgtggccagc gaagtgcggc tgctgctgag
     1861 caccaagtgg aacgtgggcc ggctgtacca gctgcccatc gtgggctctg aggaattcgc
     1921 cgagtaccac agccagcgcg aggccctgac caacgtgttc agagatctgg ccaccgagat
     1981 tgtgggcgac gaccccagag ccgaactgcc cttccacatc accatgagcg tgatcgagat
     2041 gcggcggaac gacggcaaga tccctagccc tctgagcgcc gactctctgc ccgagacagc
     2101 cattatgctg gctgacgcct ccctggctgt gctgggagca cctctgcctg ccgacagagt
     2161 ggaaaagaca ctggaactga tcaagcaggc cgacgccaag ggaggaggtg cttgtggtgc
     2221 ccaaagcgac tctgattcgt catcggtcgt tgatttcgaa ggtgggatgg agaagagatc
     2281 tcagctgtta gatctagatc ttaatttgcc tcctccatcg gaacaggccc cgaagaagaa
     2341 gaggaaggtt tgatgtcggt acgctgaaat caccagtctc tctctacaaa tctatctctc
     2401 tctattttct ccataaataa tgtgtgagta gtttcccgat aagggaaatt agggttctta
     2461 tagggtttcg ctcatgtgtt gagcatataa gaaaccctta gtatgtattt gtatttgtaa
     2521 aatacttcta tcaataaaat ttctaattcc taaaaccaaa atccagtact aaaatccaga
     2581 tcgatgcttg atactacgtc ttcgggctag agcggactag ttctagagcg gccgccaccg
     2641 cggtggagct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca
     2701 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     2761 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     2821 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     2881 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     2941 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3001 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgaa ggccttgaca
     3061 ggatatattg gcgggtaaac taagtcgctg tatgtgtttg tttgagatct catgtgagca
     3121 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3181 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3241 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3301 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3361 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3421 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3481 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3541 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3601 tacactagaa gaacagtatt tggtatctgt gctctgctga agccagttac cttcggaaga
     3661 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3721 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3781 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgaattct
     3841 tgttagacat tatttgccga ctaccttggt gatctcgcct ttcacgtagt gaacaaattc
     3901 ttccaactga tctgcgcgcg aggccaagcg atcttcttgt ccaagataag cctgcctagc
     3961 ttcaagtatg acgggctgat actgggccgg caggcgctcc attgcccagt cggcagcgac
     4021 atccttcggc gcgattttgc cggttactgc gctgtaccaa atgcgggaca acgtaagcac
     4081 tacatttcgc tcatcgccag cccagtcggg cggcgagttc catagcgtta aggtttcatt
     4141 tagcgcctca aatagatcct gttcaggaac cggatcaaag agttcctccg ccgctggacc
     4201 taccaaggca acgctatgtt ctcttgcttt tgtcagcaag atagccagat caatgtcgat
     4261 cgtggctggc tcgaagatac ctgcaagaat gtcattgcgc tgccattctc caaattgcag
     4321 ttcgcgctta gctggataac gccacggaat gatgtcgtcg tgcacaacaa tggtgacttc
     4381 tacagcgcgg agaatctcgc tctctccagg ggaagccgaa gtttccaaaa ggtcgttgat
     4441 caaagctcgc cgcgttgttt catcaagcct tacggtcacc gtaaccagca aatcaatatc
     4501 actgtgtggc ttcaggccgc catccactgc ggagccgtac aaatgtacgg ccagcaacgt
     4561 cggttcgaga tggcgctcga tgacgccaac tacctctgat agttgagtcg atacttcggc
     4621 gatcaccgct tccctcatga tgtttaactc ctgaattaag ccgcgccgcg aagcggtgtc
     4681 ggcttgaatg aattgttagg cgtcatcctg tgctcccgag aaccagtacc agtacatcgc
     4741 tgtttcgttc gagacttgag gtctagtttt atacgtgaac aggtcaatgc cgccgagagt
     4801 aaagccacat tttgcgtaca aattgcaggc aggtacattg ttcgtttgtg ccatgatgat
     4861 atatttttat cttgtgcaat gtaacatcag agattttgag acacaacgtg gctttgttga
     4921 ataaatcgaa cttttgctga gttgaaggat cagatcacgc atcttcccga caacgcagac
     4981 cgttccgtgg caaagcaaaa gttcaaaatc accaactggt ccacctacaa caaagctctc
     5041 atcaaccgtg gctccctcac tttctggctg gatgatgggg cgattcagga gatccccatc
     5101 caacagcccg ccgtcgagcg ggctttttta tccccggaag cctgtggata gagggtagtt
     5161 atccacgtga aaccgctaat gccccgcaaa gccttgattc acggggcttt ccggcccgct
     5221 ccaaaaacta tccacgtgaa atcgctaatc agggtacgtg aaatcgctaa tcggagtacg
     5281 tgaaatcgct aataaggtca cgtgaaatcg ctaatcaaaa aggcacgtga gaacgctaat
     5341 agccctttca gatcaacagc ttgcaaacac ccctcgctcc ggcaagtagt tacagcaagt
     5401 agtatgttca attagctttt caattatgaa tatatatatc aattattggt cgcccttggc
     5461 ttgtggacaa tgcgctacgc gcaccggctc cgcccgtgga caaccgcaag cggttgccca
     5521 ccgtcgagcg cctttgccca caacccggcg gccggccgca acagatcgtt ttataaattt
     5581 ttttttttga aaaagaaaaa gcccgaaagg cggcaacctc tcgggcttct ggatttccga
     5641 tccccggaat tagactctgg ctgcctgtat cgagtggtga ttttgtgccg agctgccggt
     5701 cggggagctg ttggctg
//
