LOCUS       sequence-387083-        5759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proG1090::AmtR-ZAT10(RD)-19St).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5759)
  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 5759)
  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: JAB2283
FEATURES             Location/Qualifiers
     source          1..5759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(94..346)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(366..917)
                     /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(959..1138)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     1455..1475
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        1464..1510
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             2366..2386
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2442..2618
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(2700..2720)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2700..2718)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2739..2755)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2739..2755)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2752..2774)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2763..2779
                     /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(2787..2793,2794..2811,2812..2817))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2832..2853
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2970..2987)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3092..3116
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(3207..3795)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3287..3306)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3886..4674)
                     /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      5132..5567
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(5759,1..24)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 ggcaggatat attgtggtgt aaacaaattg acgcttagac aacttaataa cacattgcgg
       61 acgtttttaa tgtaatcagc ttgcatgccg gtcgatctag taacatagat gacaccgcgc
      121 gcgataattt atcctagttt gcgcgctata ttttgttttc tatcgcgtat taaatgtata
      181 attgcgggac tctaatcata aaaacccatc tcataaataa cgtcatgcat tacatgttaa
      241 ttattacatg cttaacgtaa ttcaacagaa attatatgat aatcatcgca agaccggcaa
      301 caggattcaa tcttaagaaa ctttattgcc aaatgtttga acgatctgct tgactctagg
      361 ggtcatcaga tttcggtgac gggcaggacc ggacggggcg gcaccggcag gctgaagtcc
      421 agctgccaga aacccacgtc atgccagttc ccgtgcttga agccggccgc ccgcagcatg
      481 ccgcgggggg catatccgag cgcctcgtgc atgcgcacgc tcgggtcgtt gggcagcccg
      541 atgacagcga ccacgctctt gaagccctgt gcctccaggg acttcagcag gtgggtgtag
      601 agcgtggagc ccagtcccgt ccgctggtgg cggggggaca cgtacacggt cgactcggcc
      661 gtccagtcgt aggcgttgcg tgccttccag ggacccgcgt aggcgatgcc ggcgacctcg
      721 ccgtccacct cggcgacgag ccagggatag cgctcccgca gacggacgag gtcgtccgtc
      781 cactcctgcg gttcctgcgg ctcggtacgg aagttgaccg tgcttgtctc gatgtagtgg
      841 ttgacgatgg tgcagaccgc cggcatgtcc gcctcggtgg cacggcggat gtcggccggg
      901 cgtcgttctg ggctcatggt agatcccctc gatcgagttg agagtgaata tgagactcta
      961 attggatacc gaggggaatt tatggaacgt cagtggagca tttttgacaa gaaatatttg
     1021 ctagctgata gtgaccttag gcgacttttg aacgcgcaat aatggtttct gacgtatgtg
     1081 cttagctcat taaactccag aaacccgcgg ctcagtggct ccttcaacgt tgcggttctg
     1141 tcagttccaa acgtaaaacg gcttgtcccg cgtcatcggc gggggtcata acgtgactcc
     1201 cttaattctc cgctcatgat cgataacatg tcgactatcg gacgagctca aagtttaacg
     1261 tgtgaagacg tgcttctaat gtagtttaaa ctgaaggcgg gaaacgacaa tctgatccaa
     1321 gctcaagcta agcttgcatg cctgcaggat atcgtggatc caagcttgcc acgtgccgcc
     1381 acgtgccgcc acgtgccgcc acgtgcctct agaggatcca tctccactga cgtaagggat
     1441 gacgcacaat cccactatcc ttcgcaagac ccttcctcta tataaggaag ttcatttcat
     1501 ttggagagga cacgctggga tccccaccat ggctggcgcc gtgggcagac ccagaagatc
     1561 tgctcctcgg agagccggca agaacccccg ggaagagatt ctggatgcca gcgccgagct
     1621 gttcaccaga cagggctttg ccaccaccag cacccaccag attgccgacg ctgtgggcat
     1681 cagacaggcc agcctgtact accacttccc cagcaagacc gagatcttcc tgaccctgct
     1741 gaaaagcacc gtggaaccct ccaccgtgct ggccgaggat ctgtctaccc tggacgccgg
     1801 acccgaaatg agactgtggg ctatcgtggc cagcgaagtg cggctgctgc tgagcaccaa
     1861 gtggaacgtg ggccggctgt accagctgcc catcgtgggc tctgaggaat tcgccgagta
     1921 ccacagccag cgcgaggccc tgaccaacgt gttcagagat ctggccaccg agattgtggg
     1981 cgacgacccc agagccgaac tgcccttcca catcaccatg agcgtgatcg agatgcggcg
     2041 gaacgacggc aagatcccta gccctctgag cgccgactct ctgcccgaga cagccattat
     2101 gctggctgac gcctccctgg ctgtgctggg agcacctctg cctgccgaca gagtggaaaa
     2161 gacactggaa ctgatcaagc aggccgacgc caagggagga ggtgaaggtg cggggtccac
     2221 tagccacgtt agcagtagcc accgtgggtt tgacctcaac atccctccga tccctgaatt
     2281 ctcgatggtc aacggagacg acgaagtcat gagccctatg ccggcgaaga agcctcggtt
     2341 tgactttccg gtcaaacttc aacttccgaa gaagaagagg aaggtttagt gtcggtacgc
     2401 tgaaatcacc agtctctctc tacaaatcta tctctctcta ttttctccat aaataatgtg
     2461 tgagtagttt cccgataagg gaaattaggg ttcttatagg gtttcgctca tgtgttgagc
     2521 atataagaaa cccttagtat gtatttgtat ttgtaaaata cttctatcaa taaaatttct
     2581 aattcctaaa accaaaatcc agtactaaaa tccagatcga tgcttgatac tacgtcttcg
     2641 ggctagagcg gactagttct agagcggccg ccaccgcggt ggagctccag cttttgttcc
     2701 ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
     2761 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     2821 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     2881 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     2941 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3001 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3061 ggggataacg caggaaagaa catgaaggcc ttgacaggat atattggcgg gtaaactaag
     3121 tcgctgtatg tgtttgtttg agatctcatg tgagcaaaag gccagcaaaa ggccaggaac
     3181 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3241 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3301 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3361 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3421 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3481 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3541 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3601 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3661 atctgtgctc tgctgaagcc agttaccttc ggaagaagag ttggtagctc ttgatccggc
     3721 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3781 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3841 gaaaactcac gttaagggat tttggtcatg aattcttgtt agacattatt tgccgactac
     3901 cttggtgatc tcgcctttca cgtagtgaac aaattcttcc aactgatctg cgcgcgaggc
     3961 caagcgatct tcttgtccaa gataagcctg cctagcttca agtatgacgg gctgatactg
     4021 ggccggcagg cgctccattg cccagtcggc agcgacatcc ttcggcgcga ttttgccggt
     4081 tactgcgctg taccaaatgc gggacaacgt aagcactaca tttcgctcat cgccagccca
     4141 gtcgggcggc gagttccata gcgttaaggt ttcatttagc gcctcaaata gatcctgttc
     4201 aggaaccgga tcaaagagtt cctccgccgc tggacctacc aaggcaacgc tatgttctct
     4261 tgcttttgtc agcaagatag ccagatcaat gtcgatcgtg gctggctcga agatacctgc
     4321 aagaatgtca ttgcgctgcc attctccaaa ttgcagttcg cgcttagctg gataacgcca
     4381 cggaatgatg tcgtcgtgca caacaatggt gacttctaca gcgcggagaa tctcgctctc
     4441 tccaggggaa gccgaagttt ccaaaaggtc gttgatcaaa gctcgccgcg ttgtttcatc
     4501 aagccttacg gtcaccgtaa ccagcaaatc aatatcactg tgtggcttca ggccgccatc
     4561 cactgcggag ccgtacaaat gtacggccag caacgtcggt tcgagatggc gctcgatgac
     4621 gccaactacc tctgatagtt gagtcgatac ttcggcgatc accgcttccc tcatgatgtt
     4681 taactcctga attaagccgc gccgcgaagc ggtgtcggct tgaatgaatt gttaggcgtc
     4741 atcctgtgct cccgagaacc agtaccagta catcgctgtt tcgttcgaga cttgaggtct
     4801 agttttatac gtgaacaggt caatgccgcc gagagtaaag ccacattttg cgtacaaatt
     4861 gcaggcaggt acattgttcg tttgtgccat gatgatatat ttttatcttg tgcaatgtaa
     4921 catcagagat tttgagacac aacgtggctt tgttgaataa atcgaacttt tgctgagttg
     4981 aaggatcaga tcacgcatct tcccgacaac gcagaccgtt ccgtggcaaa gcaaaagttc
     5041 aaaatcacca actggtccac ctacaacaaa gctctcatca accgtggctc cctcactttc
     5101 tggctggatg atggggcgat tcaggagatc cccatccaac agcccgccgt cgagcgggct
     5161 tttttatccc cggaagcctg tggatagagg gtagttatcc acgtgaaacc gctaatgccc
     5221 cgcaaagcct tgattcacgg ggctttccgg cccgctccaa aaactatcca cgtgaaatcg
     5281 ctaatcaggg tacgtgaaat cgctaatcgg agtacgtgaa atcgctaata aggtcacgtg
     5341 aaatcgctaa tcaaaaaggc acgtgagaac gctaatagcc ctttcagatc aacagcttgc
     5401 aaacacccct cgctccggca agtagttaca gcaagtagta tgttcaatta gcttttcaat
     5461 tatgaatata tatatcaatt attggtcgcc cttggcttgt ggacaatgcg ctacgcgcac
     5521 cggctccgcc cgtggacaac cgcaagcggt tgcccaccgt cgagcgcctt tgcccacaac
     5581 ccggcggccg gccgcaacag atcgttttat aaattttttt ttttgaaaaa gaaaaagccc
     5641 gaaaggcggc aacctctcgg gcttctggat ttccgatccc cggaattaga ctctggctgc
     5701 ctgtatcgag tggtgatttt gtgccgagct gccggtcggg gagctgttgg ctggctggt
//
