LOCUS       sequence-389485-        5543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proG1090::TetR-NLS_19St).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5543)
  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 5543)
  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: JAB2514
FEATURES             Location/Qualifiers
     source          1..5543
                     /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             2151..2171
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2227..2403
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(2485..2505)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2485..2503)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2524..2540)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2524..2540)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2537..2559)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2548..2564
                     /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(2572..2578,2579..2596,2597..2602))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2617..2638
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2755..2772)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2877..2901
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(2992..3580)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3072..3091)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3671..4459)
                     /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      4917..5352
                     /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 tgagccggct ggacaagagc aaagtgatca
     1561 acagcgccct ggaactgctc aacgaagtgg gcatcgaggg cctgaccacc cggaagctgg
     1621 ctcagaaact gggcgtggaa cagcccaccc tgtactggca cgtgaagaac aagagagccc
     1681 tgctggacgc cctggccatc gagatgctgg accggcacca cacccacttt tgccccctgg
     1741 aaggcgagag ctggcaggat ttcctgcgga acaacgccaa gagcttcaga tgcgctctgc
     1801 tgtcccaccg ggatggcgcc aaagtgcacc tgggcaccag acctaccgag aagcagtacg
     1861 agacactgga aaaccagctg gccttcctgt gccagcaggg cttcagcctg gaaaatgccc
     1921 tgtacgccct gagcgccgtg ggccacttta cactgggctg cgtgctggaa gatcaggaac
     1981 accaggtcgc caaagaggaa agagagacac ccaccaccga cagcatgccc cctctgctga
     2041 gacaggccat tgagctgttc gatcatcagg gcgccgagcc cgcctttctg ttcggcctcg
     2101 agctgatcat ctgtggcctc gaaaaacagc tgaagtgcga gtccggcagc ccgaagaaga
     2161 agaggaaggt ttagtgtcgg tacgctgaaa tcaccagtct ctctctacaa atctatctct
     2221 ctctattttc tccataaata atgtgtgagt agtttcccga taagggaaat tagggttctt
     2281 atagggtttc gctcatgtgt tgagcatata agaaaccctt agtatgtatt tgtatttgta
     2341 aaatacttct atcaataaaa tttctaattc ctaaaaccaa aatccagtac taaaatccag
     2401 atcgatgctt gatactacgt cttcgggcta gagcggacta gttctagagc ggccgccacc
     2461 gcggtggagc tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
     2521 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     2581 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     2641 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     2701 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     2761 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2821 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatga aggccttgac
     2881 aggatatatt ggcgggtaaa ctaagtcgct gtatgtgttt gtttgagatc tcatgtgagc
     2941 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3001 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3061 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3121 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3181 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3241 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3301 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3361 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3421 ctacactaga agaacagtat ttggtatctg tgctctgctg aagccagtta ccttcggaag
     3481 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3541 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3601 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgaattc
     3661 ttgttagaca ttatttgccg actaccttgg tgatctcgcc tttcacgtag tgaacaaatt
     3721 cttccaactg atctgcgcgc gaggccaagc gatcttcttg tccaagataa gcctgcctag
     3781 cttcaagtat gacgggctga tactgggccg gcaggcgctc cattgcccag tcggcagcga
     3841 catccttcgg cgcgattttg ccggttactg cgctgtacca aatgcgggac aacgtaagca
     3901 ctacatttcg ctcatcgcca gcccagtcgg gcggcgagtt ccatagcgtt aaggtttcat
     3961 ttagcgcctc aaatagatcc tgttcaggaa ccggatcaaa gagttcctcc gccgctggac
     4021 ctaccaaggc aacgctatgt tctcttgctt ttgtcagcaa gatagccaga tcaatgtcga
     4081 tcgtggctgg ctcgaagata cctgcaagaa tgtcattgcg ctgccattct ccaaattgca
     4141 gttcgcgctt agctggataa cgccacggaa tgatgtcgtc gtgcacaaca atggtgactt
     4201 ctacagcgcg gagaatctcg ctctctccag gggaagccga agtttccaaa aggtcgttga
     4261 tcaaagctcg ccgcgttgtt tcatcaagcc ttacggtcac cgtaaccagc aaatcaatat
     4321 cactgtgtgg cttcaggccg ccatccactg cggagccgta caaatgtacg gccagcaacg
     4381 tcggttcgag atggcgctcg atgacgccaa ctacctctga tagttgagtc gatacttcgg
     4441 cgatcaccgc ttccctcatg atgtttaact cctgaattaa gccgcgccgc gaagcggtgt
     4501 cggcttgaat gaattgttag gcgtcatcct gtgctcccga gaaccagtac cagtacatcg
     4561 ctgtttcgtt cgagacttga ggtctagttt tatacgtgaa caggtcaatg ccgccgagag
     4621 taaagccaca ttttgcgtac aaattgcagg caggtacatt gttcgtttgt gccatgatga
     4681 tatattttta tcttgtgcaa tgtaacatca gagattttga gacacaacgt ggctttgttg
     4741 aataaatcga acttttgctg agttgaagga tcagatcacg catcttcccg acaacgcaga
     4801 ccgttccgtg gcaaagcaaa agttcaaaat caccaactgg tccacctaca acaaagctct
     4861 catcaaccgt ggctccctca ctttctggct ggatgatggg gcgattcagg agatccccat
     4921 ccaacagccc gccgtcgagc gggctttttt atccccggaa gcctgtggat agagggtagt
     4981 tatccacgtg aaaccgctaa tgccccgcaa agccttgatt cacggggctt tccggcccgc
     5041 tccaaaaact atccacgtga aatcgctaat cagggtacgt gaaatcgcta atcggagtac
     5101 gtgaaatcgc taataaggtc acgtgaaatc gctaatcaaa aaggcacgtg agaacgctaa
     5161 tagccctttc agatcaacag cttgcaaaca cccctcgctc cggcaagtag ttacagcaag
     5221 tagtatgttc aattagcttt tcaattatga atatatatat caattattgg tcgcccttgg
     5281 cttgtggaca atgcgctacg cgcaccggct ccgcccgtgg acaaccgcaa gcggttgccc
     5341 accgtcgagc gcctttgccc acaacccggc ggccggccgc aacagatcgt tttataaatt
     5401 tttttttttg aaaaagaaaa agcccgaaag gcggcaacct ctcgggcttc tggatttccg
     5461 atccccggaa ttagactctg gctgcctgta tcgagtggtg attttgtgcc gagctgccgg
     5521 tcggggagct gttggctggc tgg
//
