LOCUS       sequence-387078-        5831 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proG1090::AmtR-ERF2(AD)-19St).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5831)
  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 5831)
  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: JAB2281
FEATURES             Location/Qualifiers
     source          1..5831
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..25
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(95..347)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(367..918)
                     /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(960..1139)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     1456..1476
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        1465..1511
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             2439..2459
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2515..2691
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(2773..2793)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2773..2791)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2812..2828)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2812..2828)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2825..2847)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2836..2852
                     /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(2860..2866,2867..2884,2885..2890))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2905..2926
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3043..3060)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3165..3189
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(3280..3868)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3360..3379)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3959..4747)
                     /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      5205..5640
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 tggcaggata tattgtggtg taaacaaatt gacgcttaga caacttaata acacattgcg
       61 gacgttttta atgtaatcag cttgcatgcc ggtcgatcta gtaacataga tgacaccgcg
      121 cgcgataatt tatcctagtt tgcgcgctat attttgtttt ctatcgcgta ttaaatgtat
      181 aattgcggga ctctaatcat aaaaacccat ctcataaata acgtcatgca ttacatgtta
      241 attattacat gcttaacgta attcaacaga aattatatga taatcatcgc aagaccggca
      301 acaggattca atcttaagaa actttattgc caaatgtttg aacgatctgc ttgactctag
      361 gggtcatcag atttcggtga cgggcaggac cggacggggc ggcaccggca ggctgaagtc
      421 cagctgccag aaacccacgt catgccagtt cccgtgcttg aagccggccg cccgcagcat
      481 gccgcggggg gcatatccga gcgcctcgtg catgcgcacg ctcgggtcgt tgggcagccc
      541 gatgacagcg accacgctct tgaagccctg tgcctccagg gacttcagca ggtgggtgta
      601 gagcgtggag cccagtcccg tccgctggtg gcggggggac acgtacacgg tcgactcggc
      661 cgtccagtcg taggcgttgc gtgccttcca gggacccgcg taggcgatgc cggcgacctc
      721 gccgtccacc tcggcgacga gccagggata gcgctcccgc agacggacga ggtcgtccgt
      781 ccactcctgc ggttcctgcg gctcggtacg gaagttgacc gtgcttgtct cgatgtagtg
      841 gttgacgatg gtgcagaccg ccggcatgtc cgcctcggtg gcacggcgga tgtcggccgg
      901 gcgtcgttct gggctcatgg tagatcccct cgatcgagtt gagagtgaat atgagactct
      961 aattggatac cgaggggaat ttatggaacg tcagtggagc atttttgaca agaaatattt
     1021 gctagctgat agtgacctta ggcgactttt gaacgcgcaa taatggtttc tgacgtatgt
     1081 gcttagctca ttaaactcca gaaacccgcg gctcagtggc tccttcaacg ttgcggttct
     1141 gtcagttcca aacgtaaaac ggcttgtccc gcgtcatcgg cgggggtcat aacgtgactc
     1201 ccttaattct ccgctcatga tcgataacat gtcgactatc ggacgagctc aaagtttaac
     1261 gtgtgaagac gtgcttctaa tgtagtttaa actgaaggcg ggaaacgaca atctgatcca
     1321 agctcaagct aagcttgcat gcctgcagga tatcgtggat ccaagcttgc cacgtgccgc
     1381 cacgtgccgc cacgtgccgc cacgtgcctc tagaggatcc atctccactg acgtaaggga
     1441 tgacgcacaa tcccactatc cttcgcaaga cccttcctct atataaggaa gttcatttca
     1501 tttggagagg acacgctggg atccccacca tggctggcgc cgtgggcaga cccagaagat
     1561 ctgctcctcg gagagccggc aagaaccccc gggaagagat tctggatgcc agcgccgagc
     1621 tgttcaccag acagggcttt gccaccacca gcacccacca gattgccgac gctgtgggca
     1681 tcagacaggc cagcctgtac taccacttcc ccagcaagac cgagatcttc ctgaccctgc
     1741 tgaaaagcac cgtggaaccc tccaccgtgc tggccgagga tctgtctacc ctggacgccg
     1801 gacccgaaat gagactgtgg gctatcgtgg ccagcgaagt gcggctgctg ctgagcacca
     1861 agtggaacgt gggccggctg taccagctgc ccatcgtggg ctctgaggaa ttcgccgagt
     1921 accacagcca gcgcgaggcc ctgaccaacg tgttcagaga tctggccacc gagattgtgg
     1981 gcgacgaccc cagagccgaa ctgcccttcc acatcaccat gagcgtgatc gagatgcggc
     2041 ggaacgacgg caagatccct agccctctga gcgccgactc tctgcccgag acagccatta
     2101 tgctggctga cgcctccctg gctgtgctgg gagcacctct gcctgccgac agagtggaaa
     2161 agacactgga actgatcaag caggccgacg ccaagggagg aggtgaatcc gactacgctt
     2221 tgttggagtc gataacacgt cacttgctag gaggaggagg agagaacgag ctgcgactca
     2281 atgagtcaac accgagttcg tgtttcacag agagttgggg aggtttgcca ttgaaagaga
     2341 atgattcaga ggacatgttg gtgtacggac tcctcaaaga tgccttccat tttgacacgt
     2401 catcatcgga cttgagctgt ctttttgatt ttccggcgcc gaagaagaag aggaaggttt
     2461 agtgtcggta cgctgaaatc accagtctct ctctacaaat ctatctctct ctattttctc
     2521 cataaataat gtgtgagtag tttcccgata agggaaatta gggttcttat agggtttcgc
     2581 tcatgtgttg agcatataag aaacccttag tatgtatttg tatttgtaaa atacttctat
     2641 caataaaatt tctaattcct aaaaccaaaa tccagtacta aaatccagat cgatgcttga
     2701 tactacgtct tcgggctaga gcggactagt tctagagcgg ccgccaccgc ggtggagctc
     2761 cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct
     2821 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     2881 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     2941 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3001 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3061 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3121 atccacagaa tcaggggata acgcaggaaa gaacatgaag gccttgacag gatatattgg
     3181 cgggtaaact aagtcgctgt atgtgtttgt ttgagatctc atgtgagcaa aaggccagca
     3241 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3301 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3361 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3421 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3481 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3541 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3601 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3661 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3721 aacagtattt ggtatctgtg ctctgctgaa gccagttacc ttcggaagaa gagttggtag
     3781 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3841 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3901 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgaattctt gttagacatt
     3961 atttgccgac taccttggtg atctcgcctt tcacgtagtg aacaaattct tccaactgat
     4021 ctgcgcgcga ggccaagcga tcttcttgtc caagataagc ctgcctagct tcaagtatga
     4081 cgggctgata ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg
     4141 cgattttgcc ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct
     4201 catcgccagc ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa
     4261 atagatcctg ttcaggaacc ggatcaaaga gttcctccgc cgctggacct accaaggcaa
     4321 cgctatgttc tcttgctttt gtcagcaaga tagccagatc aatgtcgatc gtggctggct
     4381 cgaagatacc tgcaagaatg tcattgcgct gccattctcc aaattgcagt tcgcgcttag
     4441 ctggataacg ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga
     4501 gaatctcgct ctctccaggg gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc
     4561 gcgttgtttc atcaagcctt acggtcaccg taaccagcaa atcaatatca ctgtgtggct
     4621 tcaggccgcc atccactgcg gagccgtaca aatgtacggc cagcaacgtc ggttcgagat
     4681 ggcgctcgat gacgccaact acctctgata gttgagtcga tacttcggcg atcaccgctt
     4741 ccctcatgat gtttaactcc tgaattaagc cgcgccgcga agcggtgtcg gcttgaatga
     4801 attgttaggc gtcatcctgt gctcccgaga accagtacca gtacatcgct gtttcgttcg
     4861 agacttgagg tctagtttta tacgtgaaca ggtcaatgcc gccgagagta aagccacatt
     4921 ttgcgtacaa attgcaggca ggtacattgt tcgtttgtgc catgatgata tatttttatc
     4981 ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttgttgaa taaatcgaac
     5041 ttttgctgag ttgaaggatc agatcacgca tcttcccgac aacgcagacc gttccgtggc
     5101 aaagcaaaag ttcaaaatca ccaactggtc cacctacaac aaagctctca tcaaccgtgg
     5161 ctccctcact ttctggctgg atgatggggc gattcaggag atccccatcc aacagcccgc
     5221 cgtcgagcgg gcttttttat ccccggaagc ctgtggatag agggtagtta tccacgtgaa
     5281 accgctaatg ccccgcaaag ccttgattca cggggctttc cggcccgctc caaaaactat
     5341 ccacgtgaaa tcgctaatca gggtacgtga aatcgctaat cggagtacgt gaaatcgcta
     5401 ataaggtcac gtgaaatcgc taatcaaaaa ggcacgtgag aacgctaata gccctttcag
     5461 atcaacagct tgcaaacacc cctcgctccg gcaagtagtt acagcaagta gtatgttcaa
     5521 ttagcttttc aattatgaat atatatatca attattggtc gcccttggct tgtggacaat
     5581 gcgctacgcg caccggctcc gcccgtggac aaccgcaagc ggttgcccac cgtcgagcgc
     5641 ctttgcccac aacccggcgg ccggccgcaa cagatcgttt tataaatttt tttttttgaa
     5701 aaagaaaaag cccgaaaggc ggcaacctct cgggcttctg gatttccgat ccccggaatt
     5761 agactctggc tgcctgtatc gagtggtgat tttgtgccga gctgccggtc ggggagctgt
     5821 tggctggctg g
//
