LOCUS       sequence-387112-        5561 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proG1090::McbR-NLS-19St).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5561)
  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 5561)
  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: JAB3017
FEATURES             Location/Qualifiers
     source          1..5561
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    27..51
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(121..373)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(393..944)
                     /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(986..1165)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     1482..1502
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        1491..1537
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             2195..2215
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2271..2447
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(2529..2549)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2529..2547)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2568..2584)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2568..2584)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2581..2603)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2592..2608
                     /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(2616..2622,2623..2640,2641..2646))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2661..2682
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2799..2816)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2921..2945
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(3036..3624)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3116..3135)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3715..4503)
                     /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      4961..5396
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 cggtcgggga gctgttggct ggctggtggc aggatatatt gtggtgtaaa caaattgacg
       61 cttagacaac ttaataacac attgcggacg tttttaatgt aatcagcttg catgccggtc
      121 gatctagtaa catagatgac accgcgcgcg ataatttatc ctagtttgcg cgctatattt
      181 tgttttctat cgcgtattaa atgtataatt gcgggactct aatcataaaa acccatctca
      241 taaataacgt catgcattac atgttaatta ttacatgctt aacgtaattc aacagaaatt
      301 atatgataat catcgcaaga ccggcaacag gattcaatct taagaaactt tattgccaaa
      361 tgtttgaacg atctgcttga ctctaggggt catcagattt cggtgacggg caggaccgga
      421 cggggcggca ccggcaggct gaagtccagc tgccagaaac ccacgtcatg ccagttcccg
      481 tgcttgaagc cggccgcccg cagcatgccg cggggggcat atccgagcgc ctcgtgcatg
      541 cgcacgctcg ggtcgttggg cagcccgatg acagcgacca cgctcttgaa gccctgtgcc
      601 tccagggact tcagcaggtg ggtgtagagc gtggagccca gtcccgtccg ctggtggcgg
      661 ggggacacgt acacggtcga ctcggccgtc cagtcgtagg cgttgcgtgc cttccaggga
      721 cccgcgtagg cgatgccggc gacctcgccg tccacctcgg cgacgagcca gggatagcgc
      781 tcccgcagac ggacgaggtc gtccgtccac tcctgcggtt cctgcggctc ggtacggaag
      841 ttgaccgtgc ttgtctcgat gtagtggttg acgatggtgc agaccgccgg catgtccgcc
      901 tcggtggcac ggcggatgtc ggccgggcgt cgttctgggc tcatggtaga tcccctcgat
      961 cgagttgaga gtgaatatga gactctaatt ggataccgag gggaatttat ggaacgtcag
     1021 tggagcattt ttgacaagaa atatttgcta gctgatagtg accttaggcg acttttgaac
     1081 gcgcaataat ggtttctgac gtatgtgctt agctcattaa actccagaaa cccgcggctc
     1141 agtggctcct tcaacgttgc ggttctgtca gttccaaacg taaaacggct tgtcccgcgt
     1201 catcggcggg ggtcataacg tgactccctt aattctccgc tcatgatcga taacatgtcg
     1261 actatcggac gagctcaaag tttaacgtgt gaagacgtgc ttctaatgta gtttaaactg
     1321 aaggcgggaa acgacaatct gatccaagct caagctaagc ttgcatgcct gcaggatatc
     1381 gtggatccaa gcttgccacg tgccgccacg tgccgccacg tgccgccacg tgcctctaga
     1441 ggatccatct ccactgacgt aagggatgac gcacaatccc actatccttc gcaagaccct
     1501 tcctctatat aaggaagttc atttcatttg gagaggacac gctgggatcc ccaccatggc
     1561 cgccagcgcc agcggcaaga gcaaaacaag cgctggcgcc aaccggcgga gaaacagacc
     1621 cagccctaga cagagactgc tggacagcgc caccaacctg ttcaccaccg agggcatcag
     1681 agtgatcggc atcgaccgga tcctgcgcga ggccgatgtg gccaaggcca gcctgtacag
     1741 cctgttcggc agcaaggatg ccctcgtgat cgcctacctg gaaaacctgg accagctgtg
     1801 gcgcgaagcc tggcgggaaa gaaccgtggg catgaaggac cccgaggaca agattatcgc
     1861 cttcttcgac cagtgcatcg aggaagaacc cgagaaggac ttccggggca gccacttcca
     1921 gaacgccgcc tccgagtacc ccagacccga aaccgatagc gagaagggca ttgtggccgc
     1981 cgtgctggaa cacagagagt ggtgccacaa gaccctgacc gacctgctga ccgagaagaa
     2041 cggctaccct ggcaccacac aggccaacca gctgctggtg tttctggatg gcggcctggc
     2101 cggcagcaga ctggtgcaca atatcagccc cctggaaacc gccagagatc tggccagaca
     2161 gctgctctct gccccccctg ccgactactc tatcccgaag aagaagagga aggtttagtg
     2221 tcggtacgct gaaatcacca gtctctctct acaaatctat ctctctctat tttctccata
     2281 aataatgtgt gagtagtttc ccgataaggg aaattagggt tcttataggg tttcgctcat
     2341 gtgttgagca tataagaaac ccttagtatg tatttgtatt tgtaaaatac ttctatcaat
     2401 aaaatttcta attcctaaaa ccaaaatcca gtactaaaat ccagatcgat gcttgatact
     2461 acgtcttcgg gctagagcgg actagttcta gagcggccgc caccgcggtg gagctccagc
     2521 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
     2581 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     2641 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     2701 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     2761 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     2821 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     2881 acagaatcag gggataacgc aggaaagaac atgaaggcct tgacaggata tattggcggg
     2941 taaactaagt cgctgtatgt gtttgtttga gatctcatgt gagcaaaagg ccagcaaaag
     3001 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3061 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3121 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3181 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3241 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3301 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3361 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3421 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3481 gtatttggta tctgtgctct gctgaagcca gttaccttcg gaagaagagt tggtagctct
     3541 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3601 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3661 cagtggaacg aaaactcacg ttaagggatt ttggtcatga attcttgtta gacattattt
     3721 gccgactacc ttggtgatct cgcctttcac gtagtgaaca aattcttcca actgatctgc
     3781 gcgcgaggcc aagcgatctt cttgtccaag ataagcctgc ctagcttcaa gtatgacggg
     3841 ctgatactgg gccggcaggc gctccattgc ccagtcggca gcgacatcct tcggcgcgat
     3901 tttgccggtt actgcgctgt accaaatgcg ggacaacgta agcactacat ttcgctcatc
     3961 gccagcccag tcgggcggcg agttccatag cgttaaggtt tcatttagcg cctcaaatag
     4021 atcctgttca ggaaccggat caaagagttc ctccgccgct ggacctacca aggcaacgct
     4081 atgttctctt gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa
     4141 gatacctgca agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg
     4201 ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg acttctacag cgcggagaat
     4261 ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt
     4321 tgtttcatca agccttacgg tcaccgtaac cagcaaatca atatcactgt gtggcttcag
     4381 gccgccatcc actgcggagc cgtacaaatg tacggccagc aacgtcggtt cgagatggcg
     4441 ctcgatgacg ccaactacct ctgatagttg agtcgatact tcggcgatca ccgcttccct
     4501 catgatgttt aactcctgaa ttaagccgcg ccgcgaagcg gtgtcggctt gaatgaattg
     4561 ttaggcgtca tcctgtgctc ccgagaacca gtaccagtac atcgctgttt cgttcgagac
     4621 ttgaggtcta gttttatacg tgaacaggtc aatgccgccg agagtaaagc cacattttgc
     4681 gtacaaattg caggcaggta cattgttcgt ttgtgccatg atgatatatt tttatcttgt
     4741 gcaatgtaac atcagagatt ttgagacaca acgtggcttt gttgaataaa tcgaactttt
     4801 gctgagttga aggatcagat cacgcatctt cccgacaacg cagaccgttc cgtggcaaag
     4861 caaaagttca aaatcaccaa ctggtccacc tacaacaaag ctctcatcaa ccgtggctcc
     4921 ctcactttct ggctggatga tggggcgatt caggagatcc ccatccaaca gcccgccgtc
     4981 gagcgggctt ttttatcccc ggaagcctgt ggatagaggg tagttatcca cgtgaaaccg
     5041 ctaatgcccc gcaaagcctt gattcacggg gctttccggc ccgctccaaa aactatccac
     5101 gtgaaatcgc taatcagggt acgtgaaatc gctaatcgga gtacgtgaaa tcgctaataa
     5161 ggtcacgtga aatcgctaat caaaaaggca cgtgagaacg ctaatagccc tttcagatca
     5221 acagcttgca aacacccctc gctccggcaa gtagttacag caagtagtat gttcaattag
     5281 cttttcaatt atgaatatat atatcaatta ttggtcgccc ttggcttgtg gacaatgcgc
     5341 tacgcgcacc ggctccgccc gtggacaacc gcaagcggtt gcccaccgtc gagcgccttt
     5401 gcccacaacc cggcggccgg ccgcaacaga tcgttttata aatttttttt tttgaaaaag
     5461 aaaaagcccg aaaggcggca acctctcggg cttctggatt tccgatcccc ggaattagac
     5521 tctggctgcc tgtatcgagt ggtgattttg tgccgagctg c
//
