LOCUS       sequence-387086-        5765 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proG1090::PhlF-ERF2(AD)-19St).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5765)
  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 5765)
  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: JAB2337
FEATURES             Location/Qualifiers
     source          1..5765
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    32..56
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(126..378)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(398..949)
                     /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(991..1170)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     1487..1507
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        1496..1542
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             2404..2424
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2480..2656
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(2738..2758)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2738..2756)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2777..2793)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2777..2793)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2790..2812)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2801..2817
                     /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(2825..2831,2832..2849,2850..2855))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2870..2891
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3008..3025)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3130..3154
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(3245..3833)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3325..3344)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3924..4712)
                     /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      5170..5605
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 gctgccggtc ggggagctgt tggctggctg gtggcaggat atattgtggt gtaaacaaat
       61 tgacgcttag acaacttaat aacacattgc ggacgttttt aatgtaatca gcttgcatgc
      121 cggtcgatct agtaacatag atgacaccgc gcgcgataat ttatcctagt ttgcgcgcta
      181 tattttgttt tctatcgcgt attaaatgta taattgcggg actctaatca taaaaaccca
      241 tctcataaat aacgtcatgc attacatgtt aattattaca tgcttaacgt aattcaacag
      301 aaattatatg ataatcatcg caagaccggc aacaggattc aatcttaaga aactttattg
      361 ccaaatgttt gaacgatctg cttgactcta ggggtcatca gatttcggtg acgggcagga
      421 ccggacgggg cggcaccggc aggctgaagt ccagctgcca gaaacccacg tcatgccagt
      481 tcccgtgctt gaagccggcc gcccgcagca tgccgcgggg ggcatatccg agcgcctcgt
      541 gcatgcgcac gctcgggtcg ttgggcagcc cgatgacagc gaccacgctc ttgaagccct
      601 gtgcctccag ggacttcagc aggtgggtgt agagcgtgga gcccagtccc gtccgctggt
      661 ggcgggggga cacgtacacg gtcgactcgg ccgtccagtc gtaggcgttg cgtgccttcc
      721 agggacccgc gtaggcgatg ccggcgacct cgccgtccac ctcggcgacg agccagggat
      781 agcgctcccg cagacggacg aggtcgtccg tccactcctg cggttcctgc ggctcggtac
      841 ggaagttgac cgtgcttgtc tcgatgtagt ggttgacgat ggtgcagacc gccggcatgt
      901 ccgcctcggt ggcacggcgg atgtcggccg ggcgtcgttc tgggctcatg gtagatcccc
      961 tcgatcgagt tgagagtgaa tatgagactc taattggata ccgaggggaa tttatggaac
     1021 gtcagtggag catttttgac aagaaatatt tgctagctga tagtgacctt aggcgacttt
     1081 tgaacgcgca ataatggttt ctgacgtatg tgcttagctc attaaactcc agaaacccgc
     1141 ggctcagtgg ctccttcaac gttgcggttc tgtcagttcc aaacgtaaaa cggcttgtcc
     1201 cgcgtcatcg gcgggggtca taacgtgact cccttaattc tccgctcatg atcgataaca
     1261 tgtcgactat cggacgagct caaagtttaa cgtgtgaaga cgtgcttcta atgtagttta
     1321 aactgaaggc gggaaacgac aatctgatcc aagctcaagc taagcttgca tgcctgcagg
     1381 atatcgtgga tccaagcttg ccacgtgccg ccacgtgccg ccacgtgccg ccacgtgcct
     1441 ctagaggatc catctccact gacgtaaggg atgacgcaca atcccactat ccttcgcaag
     1501 acccttcctc tatataagga agttcatttc atttggagag gacacgctgg gatccccacc
     1561 atggcccgga cccccagcag aagcagcatc ggatctctga gaagccccca cacccacaag
     1621 gccatcctga ccagcaccat cgagatcctg aaagagtgcg gctacagcgg cctgagcatc
     1681 gagtctgtgg ctagaagggc tggcgcctcc aagcccacca tctaccggtg gtggacaaac
     1741 aaggccgccc tgatcgccga ggtgtacgag aacgagagcg agcaggtccg aaagttcccc
     1801 gacctgggca gcttcaaggc cgatctggac ttcctgctgc ggaacctgtg gaaagtgtgg
     1861 cgggaaacca tctgcggcga ggccttcaga tgcgtgatcg ctgaggccca gctggaccct
     1921 gccacactga cccagctgaa ggaccagttc atggaacggc ggagagagat gcccaagaaa
     1981 ctggtggaaa acgccatcag caacggcgag ctgcccaagg acaccaaccg ggaactgctg
     2041 ctggatatga tcttcggctt ctgctggtac agactgctga ccgagcagct gaccgtggaa
     2101 caggatatcg aggaattcac ctttctgctg atcaacggcg tgtgccccgg cacccagaga
     2161 ggaggaggtg aatccgacta cgctttgttg gagtcgataa cacgtcactt gctaggagga
     2221 ggaggagaga acgagctgcg actcaatgag tcaacaccga gttcgtgttt cacagagagt
     2281 tggggaggtt tgccattgaa agagaatgat tcagaggaca tgttggtgta cggactcctc
     2341 aaagatgcct tccattttga cacgtcatca tcggacttga gctgtctttt tgattttccg
     2401 gcgccgaaga agaagaggaa ggtttagtgt cggtacgctg aaatcaccag tctctctcta
     2461 caaatctatc tctctctatt ttctccataa ataatgtgtg agtagtttcc cgataaggga
     2521 aattagggtt cttatagggt ttcgctcatg tgttgagcat ataagaaacc cttagtatgt
     2581 atttgtattt gtaaaatact tctatcaata aaatttctaa ttcctaaaac caaaatccag
     2641 tactaaaatc cagatcgatg cttgatacta cgtcttcggg ctagagcgga ctagttctag
     2701 agcggccgcc accgcggtgg agctccagct tttgttccct ttagtgaggg ttaattgcgc
     2761 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     2821 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     2881 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     2941 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     3001 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3061 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3121 tgaaggcctt gacaggatat attggcgggt aaactaagtc gctgtatgtg tttgtttgag
     3181 atctcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3241 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3301 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3361 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3421 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3481 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3541 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3601 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3661 ggcctaacta cggctacact agaagaacag tatttggtat ctgtgctctg ctgaagccag
     3721 ttaccttcgg aagaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3781 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3841 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3901 tggtcatgaa ttcttgttag acattatttg ccgactacct tggtgatctc gcctttcacg
     3961 tagtgaacaa attcttccaa ctgatctgcg cgcgaggcca agcgatcttc ttgtccaaga
     4021 taagcctgcc tagcttcaag tatgacgggc tgatactggg ccggcaggcg ctccattgcc
     4081 cagtcggcag cgacatcctt cggcgcgatt ttgccggtta ctgcgctgta ccaaatgcgg
     4141 gacaacgtaa gcactacatt tcgctcatcg ccagcccagt cgggcggcga gttccatagc
     4201 gttaaggttt catttagcgc ctcaaataga tcctgttcag gaaccggatc aaagagttcc
     4261 tccgccgctg gacctaccaa ggcaacgcta tgttctcttg cttttgtcag caagatagcc
     4321 agatcaatgt cgatcgtggc tggctcgaag atacctgcaa gaatgtcatt gcgctgccat
     4381 tctccaaatt gcagttcgcg cttagctgga taacgccacg gaatgatgtc gtcgtgcaca
     4441 acaatggtga cttctacagc gcggagaatc tcgctctctc caggggaagc cgaagtttcc
     4501 aaaaggtcgt tgatcaaagc tcgccgcgtt gtttcatcaa gccttacggt caccgtaacc
     4561 agcaaatcaa tatcactgtg tggcttcagg ccgccatcca ctgcggagcc gtacaaatgt
     4621 acggccagca acgtcggttc gagatggcgc tcgatgacgc caactacctc tgatagttga
     4681 gtcgatactt cggcgatcac cgcttccctc atgatgttta actcctgaat taagccgcgc
     4741 cgcgaagcgg tgtcggcttg aatgaattgt taggcgtcat cctgtgctcc cgagaaccag
     4801 taccagtaca tcgctgtttc gttcgagact tgaggtctag ttttatacgt gaacaggtca
     4861 atgccgccga gagtaaagcc acattttgcg tacaaattgc aggcaggtac attgttcgtt
     4921 tgtgccatga tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacaa
     4981 cgtggctttg ttgaataaat cgaacttttg ctgagttgaa ggatcagatc acgcatcttc
     5041 ccgacaacgc agaccgttcc gtggcaaagc aaaagttcaa aatcaccaac tggtccacct
     5101 acaacaaagc tctcatcaac cgtggctccc tcactttctg gctggatgat ggggcgattc
     5161 aggagatccc catccaacag cccgccgtcg agcgggcttt tttatccccg gaagcctgtg
     5221 gatagagggt agttatccac gtgaaaccgc taatgccccg caaagccttg attcacgggg
     5281 ctttccggcc cgctccaaaa actatccacg tgaaatcgct aatcagggta cgtgaaatcg
     5341 ctaatcggag tacgtgaaat cgctaataag gtcacgtgaa atcgctaatc aaaaaggcac
     5401 gtgagaacgc taatagccct ttcagatcaa cagcttgcaa acacccctcg ctccggcaag
     5461 tagttacagc aagtagtatg ttcaattagc ttttcaatta tgaatatata tatcaattat
     5521 tggtcgccct tggcttgtgg acaatgcgct acgcgcaccg gctccgcccg tggacaaccg
     5581 caagcggttg cccaccgtcg agcgcctttg cccacaaccc ggcggccggc cgcaacagat
     5641 cgttttataa attttttttt ttgaaaaaga aaaagcccga aaggcggcaa cctctcgggc
     5701 ttctggattt ccgatccccg gaattagact ctggctgcct gtatcgagtg gtgattttgt
     5761 gccga
//
