LOCUS       sequence-387097-        5507 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proG1090::BetI-NLS_19St).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5507)
  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 5507)
  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: JAB2519
FEATURES             Location/Qualifiers
     source          1..5507
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(87..339)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(359..910)
                     /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(952..1131)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     1448..1468
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        1457..1503
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             2107..2127
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2183..2359
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(2441..2461)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2441..2459)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2480..2496)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2480..2496)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2493..2515)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2504..2520
                     /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(2528..2534,2535..2552,2553..2558))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2573..2594
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2711..2728)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2833..2857
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(2948..3536)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3028..3047)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3627..4415)
                     /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      4873..5308
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(5500..5507,1..17)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 tatattgtgg tgtaaacaaa ttgacgctta gacaacttaa taacatattg cggacgtttt
       61 taatgtaatc agcttgcatg ccggtcgatc tagtaacata gatgacaccg cgcgcgataa
      121 tttatcctag tttgcgcgct atattttgtt ttctatcgcg tattaaatgt ataattgcgg
      181 gactctaatc ataaaaaccc atctcataaa taacgtcatg cattacatgt taattattac
      241 atgcttaacg taattcaaca gaaattatat gataatcatc gcaagaccgg caacaggatt
      301 caatcttaag aaactttatt gccaaatgtt tgaacgatct gcttgactct aggggtcatc
      361 agatttcggt gacgggcagg accggacggg gcggcaccgg caggctgaag tccagctgcc
      421 agaaacccac gtcatgccag ttcccgtgct tgaagccggc cgcccgcagc atgccgcggg
      481 gggcatatcc gagcgcctcg tgcatgcgca cgctcgggtc gttgggcagc ccgatgacag
      541 cgaccacgct cttgaagccc tgtgcctcca gggacttcag caggtgggtg tagagcgtgg
      601 agcccagtcc cgtccgctgg tggcgggggg acacgtacac ggtcgactcg gccgtccagt
      661 cgtaggcgtt gcgtgccttc cagggacccg cgtaggcgat gccggcgacc tcgccgtcca
      721 cctcggcgac gagccaggga tagcgctccc gcagacggac gaggtcgtcc gtccactcct
      781 gcggttcctg cggctcggta cggaagttga ccgtgcttgt ctcgatgtag tggttgacga
      841 tggtgcagac cgccggcatg tccgcctcgg tggcacggcg gatgtcggcc gggcgtcgtt
      901 ctgggctcat ggtagatccc ctcgatcgag ttgagagtga atatgagact ctaattggat
      961 accgagggga atttatggaa cgtcagtgga gcatttttga caagaaatat ttgctagctg
     1021 atagtgacct taggcgactt ttgaacgcgc aataatggtt tctgacgtat gtgcttagct
     1081 cattaaactc cagaaacccg cggctcagtg gctccttcaa cgttgcggtt ctgtcagttc
     1141 caaacgtaaa acggcttgtc ccgcgtcatc ggcgggggtc ataacgtgac tcccttaatt
     1201 ctccgctcat gatcgataac atgtcgacta tcggacgagc tcaaagttta acgtgtgaag
     1261 acgtgcttct aatgtagttt aaactgaagg cgggaaacga caatctgatc caagctcaag
     1321 ctaagcttgc atgcctgcag gatatcgtgg atccaagctt gccacgtgcc gccacgtgcc
     1381 gccacgtgcc gccacgtgcc tctagaggat ccatctccac tgacgtaagg gatgacgcac
     1441 aatcccacta tccttcgcaa gacccttcct ctatataagg aagttcattt catttggaga
     1501 ggacacgctg ggatccccac catgccgaaa ctgggtatgc agagcattcg tcgtcgtcag
     1561 ctgattgatg caaccctgga agcaattaat gaagttggta tgcatgatgc aaccattgca
     1621 cagattgcac gtcgtgccgg tgttagcacc ggtattatta gccattattt ccgcgataaa
     1681 aacggtctgc tggaagcaac catgcgtgat attaccagcc agctgcgtga tgcagttctg
     1741 aatcgtctgc atgcactgcc gcagggtagc gcagaacagc gtctgcaggc aattgttggt
     1801 ggtaattttg atgaaaccca ggttagcagc gcagcaatga aagcatggct ggcattttgg
     1861 gcaagcagca tgcatcagcc gatgctgtat cgtctgcagc aggttagcag tcgtcgtctg
     1921 ctgagcaatc tggttagcga atttcgtcgt gaactgcctc gtgaacaggc acaagaggca
     1981 ggttatggtc tggcagcact gattgatggt ctgtggctgc gtgcagcact gagcggtaaa
     2041 ccgctggata aaacccgtgc aaatagcctg acccgtcatt ttatcaccca gcatctgccg
     2101 accgatccga agaagaagag gaaggtttag tgtcggtacg ctgaaatcac cagtctctct
     2161 ctacaaatct atctctctct attttctcca taaataatgt gtgagtagtt tcccgataag
     2221 ggaaattagg gttcttatag ggtttcgctc atgtgttgag catataagaa acccttagta
     2281 tgtatttgta tttgtaaaat acttctatca ataaaatttc taattcctaa aaccaaaatc
     2341 cagtactaaa atccagatcg atgcttgata ctacgtcttc gggctagagc ggactagttc
     2401 tagagcggcc gccaccgcgg tggagctcca gcttttgttc cctttagtga gggttaattg
     2461 cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     2521 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     2581 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     2641 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2701 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2761 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2821 acatgaaggc cttgacagga tatattggcg ggtaaactaa gtcgctgtat gtgtttgttt
     2881 gagatctcat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2941 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3001 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3061 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3121 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3181 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3241 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3301 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3361 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgtgct ctgctgaagc
     3421 cagttacctt cggaagaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3481 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3541 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3601 ttttggtcat gaattcttgt tagacattat ttgccgacta ccttggtgat ctcgcctttc
     3661 acgtagtgaa caaattcttc caactgatct gcgcgcgagg ccaagcgatc ttcttgtcca
     3721 agataagcct gcctagcttc aagtatgacg ggctgatact gggccggcag gcgctccatt
     3781 gcccagtcgg cagcgacatc cttcggcgcg attttgccgg ttactgcgct gtaccaaatg
     3841 cgggacaacg taagcactac atttcgctca tcgccagccc agtcgggcgg cgagttccat
     3901 agcgttaagg tttcatttag cgcctcaaat agatcctgtt caggaaccgg atcaaagagt
     3961 tcctccgccg ctggacctac caaggcaacg ctatgttctc ttgcttttgt cagcaagata
     4021 gccagatcaa tgtcgatcgt ggctggctcg aagatacctg caagaatgtc attgcgctgc
     4081 cattctccaa attgcagttc gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc
     4141 acaacaatgg tgacttctac agcgcggaga atctcgctct ctccagggga agccgaagtt
     4201 tccaaaaggt cgttgatcaa agctcgccgc gttgtttcat caagccttac ggtcaccgta
     4261 accagcaaat caatatcact gtgtggcttc aggccgccat ccactgcgga gccgtacaaa
     4321 tgtacggcca gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac ctctgatagt
     4381 tgagtcgata cttcggcgat caccgcttcc ctcatgatgt ttaactcctg aattaagccg
     4441 cgccgcgaag cggtgtcggc ttgaatgaat tgttaggcgt catcctgtgc tcccgagaac
     4501 cagtaccagt acatcgctgt ttcgttcgag acttgaggtc tagttttata cgtgaacagg
     4561 tcaatgccgc cgagagtaaa gccacatttt gcgtacaaat tgcaggcagg tacattgttc
     4621 gtttgtgcca tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca
     4681 caacgtggct ttgttgaata aatcgaactt ttgctgagtt gaaggatcag atcacgcatc
     4741 ttcccgacaa cgcagaccgt tccgtggcaa agcaaaagtt caaaatcacc aactggtcca
     4801 cctacaacaa agctctcatc aaccgtggct ccctcacttt ctggctggat gatggggcga
     4861 ttcaggagat ccccatccaa cagcccgccg tcgagcgggc ttttttatcc ccggaagcct
     4921 gtggatagag ggtagttatc cacgtgaaac cgctaatgcc ccgcaaagcc ttgattcacg
     4981 gggctttccg gcccgctcca aaaactatcc acgtgaaatc gctaatcagg gtacgtgaaa
     5041 tcgctaatcg gagtacgtga aatcgctaat aaggtcacgt gaaatcgcta atcaaaaagg
     5101 cacgtgagaa cgctaatagc cctttcagat caacagcttg caaacacccc tcgctccggc
     5161 aagtagttac agcaagtagt atgttcaatt agcttttcaa ttatgaatat atatatcaat
     5221 tattggtcgc ccttggcttg tggacaatgc gctacgcgca ccggctccgc ccgtggacaa
     5281 ccgcaagcgg ttgcccaccg tcgagcgcct ttgcccacaa cccggcggcc ggccgcaaca
     5341 gatcgtttta taaatttttt tttttgaaaa agaaaaagcc cgaaaggcgg caacctctcg
     5401 ggcttctgga tttccgatcc ccggaattag actctggctg cctgtatcga gtggtgattt
     5461 tgtgccgagc tgccggtcgg ggagctgttg gctggctggt ggcagga
//
