LOCUS       sequence-393579-        5675 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (6XPhlFO-3XAmtRO::TetR-NLS-19St).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5675)
  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 5675)
  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: JAB3477
FEATURES             Location/Qualifiers
     source          1..5675
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    14..38
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(108..360)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(380..931)
                     /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(973..1152)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             2296..2316
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2372..2548
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(2630..2650)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2630..2648)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2669..2685)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2669..2685)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2682..2704)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2693..2709
                     /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(2717..2723,2724..2741,2742..2747))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2762..2783
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2900..2917)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3022..3046
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(3137..3725)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3217..3236)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3816..4604)
                     /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      5062..5497
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 gttggctggc tggtggcagg atatattgtg gtgtaaacaa attgacgctt agacaactta
       61 ataacacatt gcggacgttt ttaatgtaat cagcttgcat gccggtcgat ctagtaacat
      121 agatgacacc gcgcgcgata atttatccta gtttgcgcgc tatattttgt tttctatcgc
      181 gtattaaatg tataattgcg ggactctaat cataaaaacc catctcataa ataacgtcat
      241 gcattacatg ttaattatta catgcttaac gtaattcaac agaaattata tgataatcat
      301 cgcaagaccg gcaacaggat tcaatcttaa gaaactttat tgccaaatgt ttgaacgatc
      361 tgcttgactc taggggtcat cagatttcgg tgacgggcag gaccggacgg ggcggcaccg
      421 gcaggctgaa gtccagctgc cagaaaccca cgtcatgcca gttcccgtgc ttgaagccgg
      481 ccgcccgcag catgccgcgg ggggcatatc cgagcgcctc gtgcatgcgc acgctcgggt
      541 cgttgggcag cccgatgaca gcgaccacgc tcttgaagcc ctgtgcctcc agggacttca
      601 gcaggtgggt gtagagcgtg gagcccagtc ccgtccgctg gtggcggggg gacacgtaca
      661 cggtcgactc ggccgtccag tcgtaggcgt tgcgtgcctt ccagggaccc gcgtaggcga
      721 tgccggcgac ctcgccgtcc acctcggcga cgagccaggg atagcgctcc cgcagacgga
      781 cgaggtcgtc cgtccactcc tgcggttcct gcggctcggt acggaagttg accgtgcttg
      841 tctcgatgta gtggttgacg atggtgcaga ccgccggcat gtccgcctcg gtggcacggc
      901 ggatgtcggc cgggcgtcgt tctgggctca tggtagatcc cctcgatcga gttgagagtg
      961 aatatgagac tctaattgga taccgagggg aatttatgga acgtcagtgg agcatttttg
     1021 acaagaaata tttgctagct gatagtgacc ttaggcgact tttgaacgcg caataatggt
     1081 ttctgacgta tgtgcttagc tcattaaact ccagaaaccc gcggctcagt ggctccttca
     1141 acgttgcggt tctgtcagtt ccaaacgtaa aacggcttgt cccgcgtcat cggcgggggt
     1201 cataacgtga ctcccttaat tctccgctca tgatcgataa catgtcgact atcggacgag
     1261 ctcaaagttt aacgtgtgaa gacgtgcttc taatgataga aaagttggct gcggccgcga
     1321 aatgatacga aacgtaccgt atcgttaagg tgaatgatac gaaacgtacc gtatcgttaa
     1381 ggtcaatgat acgaaacgta ccgtatcgtt aaggtcaatg atacgaaacg taccgtatcg
     1441 ttaaggtgta tgatacgaaa cgtaccgtat cgttaaggta gatgatacga aacgtaccgt
     1501 atcgttaagg tcgggtctaa gctagctagg ctagcgcaag tggattgatg tgatatctcc
     1561 actgacgtaa gggatgacgc acaatcccac tatcttctat cgatctatag ataatctata
     1621 taattctatc gatctataga taatcgttct atcgatctat agataataga caccatgagc
     1681 cggctggaca agagcaaagt gatcaacagc gccctggaac tgctcaacga agtgggcatc
     1741 gagggcctga ccacccggaa gctggctcag aaactgggcg tggaacagcc caccctgtac
     1801 tggcacgtga agaacaagag agccctgctg gacgccctgg ccatcgagat gctggaccgg
     1861 caccacaccc acttttgccc cctggaaggc gagagctggc aggatttcct gcggaacaac
     1921 gccaagagct tcagatgcgc tctgctgtcc caccgggatg gcgccaaagt gcacctgggc
     1981 accagaccta ccgagaagca gtacgagaca ctggaaaacc agctggcctt cctgtgccag
     2041 cagggcttca gcctggaaaa tgccctgtac gccctgagcg ccgtgggcca ctttacactg
     2101 ggctgcgtgc tggaagatca ggaacaccag gtcgccaaag aggaaagaga gacacccacc
     2161 accgacagca tgccccctct gctgagacag gccattgagc tgttcgatca tcagggcgcc
     2221 gagcccgcct ttctgttcgg cctcgagctg atcatctgtg gcctcgaaaa acagctgaag
     2281 tgcgagtccg gcagcccgaa gaagaagagg aaggtttagt gtcggtacgc tgaaatcacc
     2341 agtctctctc tacaaatcta tctctctcta ttttctccat aaataatgtg tgagtagttt
     2401 cccgataagg gaaattaggg ttcttatagg gtttcgctca tgtgttgagc atataagaaa
     2461 cccttagtat gtatttgtat ttgtaaaata cttctatcaa taaaatttct aattcctaaa
     2521 accaaaatcc agtactaaaa tccagatcga tgcttgatac tacgtcttcg ggctagagcg
     2581 gactagttct agagcggccg ccaccgcggt ggagctccag cttttgttcc ctttagtgag
     2641 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     2701 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     2761 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     2821 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     2881 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2941 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3001 caggaaagaa catgaaggcc ttgacaggat atattggcgg gtaaactaag tcgctgtatg
     3061 tgtttgtttg agatctcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3121 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3181 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3241 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3301 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3361 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3421 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3481 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3541 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgtgctc
     3601 tgctgaagcc agttaccttc ggaagaagag ttggtagctc ttgatccggc aaacaaacca
     3661 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3721 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3781 gttaagggat tttggtcatg aattcttgtt agacattatt tgccgactac cttggtgatc
     3841 tcgcctttca cgtagtgaac aaattcttcc aactgatctg cgcgcgaggc caagcgatct
     3901 tcttgtccaa gataagcctg cctagcttca agtatgacgg gctgatactg ggccggcagg
     3961 cgctccattg cccagtcggc agcgacatcc ttcggcgcga ttttgccggt tactgcgctg
     4021 taccaaatgc gggacaacgt aagcactaca tttcgctcat cgccagccca gtcgggcggc
     4081 gagttccata gcgttaaggt ttcatttagc gcctcaaata gatcctgttc aggaaccgga
     4141 tcaaagagtt cctccgccgc tggacctacc aaggcaacgc tatgttctct tgcttttgtc
     4201 agcaagatag ccagatcaat gtcgatcgtg gctggctcga agatacctgc aagaatgtca
     4261 ttgcgctgcc attctccaaa ttgcagttcg cgcttagctg gataacgcca cggaatgatg
     4321 tcgtcgtgca caacaatggt gacttctaca gcgcggagaa tctcgctctc tccaggggaa
     4381 gccgaagttt ccaaaaggtc gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg
     4441 gtcaccgtaa ccagcaaatc aatatcactg tgtggcttca ggccgccatc cactgcggag
     4501 ccgtacaaat gtacggccag caacgtcggt tcgagatggc gctcgatgac gccaactacc
     4561 tctgatagtt gagtcgatac ttcggcgatc accgcttccc tcatgatgtt taactcctga
     4621 attaagccgc gccgcgaagc ggtgtcggct tgaatgaatt gttaggcgtc atcctgtgct
     4681 cccgagaacc agtaccagta catcgctgtt tcgttcgaga cttgaggtct agttttatac
     4741 gtgaacaggt caatgccgcc gagagtaaag ccacattttg cgtacaaatt gcaggcaggt
     4801 acattgttcg tttgtgccat gatgatatat ttttatcttg tgcaatgtaa catcagagat
     4861 tttgagacac aacgtggctt tgttgaataa atcgaacttt tgctgagttg aaggatcaga
     4921 tcacgcatct tcccgacaac gcagaccgtt ccgtggcaaa gcaaaagttc aaaatcacca
     4981 actggtccac ctacaacaaa gctctcatca accgtggctc cctcactttc tggctggatg
     5041 atggggcgat tcaggagatc cccatccaac agcccgccgt cgagcgggct tttttatccc
     5101 cggaagcctg tggatagagg gtagttatcc acgtgaaacc gctaatgccc cgcaaagcct
     5161 tgattcacgg ggctttccgg cccgctccaa aaactatcca cgtgaaatcg ctaatcaggg
     5221 tacgtgaaat cgctaatcgg agtacgtgaa atcgctaata aggtcacgtg aaatcgctaa
     5281 tcaaaaaggc acgtgagaac gctaatagcc ctttcagatc aacagcttgc aaacacccct
     5341 cgctccggca agtagttaca gcaagtagta tgttcaatta gcttttcaat tatgaatata
     5401 tatatcaatt attggtcgcc cttggcttgt ggacaatgcg ctacgcgcac cggctccgcc
     5461 cgtggacaac cgcaagcggt tgcccaccgt cgagcgcctt tgcccacaac ccggcggccg
     5521 gccgcaacag atcgttttat aaattttttt ttttgaaaaa gaaaaagccc gaaaggcggc
     5581 aacctctcgg gcttctggat ttccgatccc cggaattaga ctctggctgc ctgtatcgag
     5641 tggtgatttt gtgccgagct gccggtcggg gagct
//
