LOCUS       sequence-393422-        5487 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proG1090::QacR-NLS_19St).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5487)
  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 5487)
  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: JAB2497
FEATURES             Location/Qualifiers
     source          1..5487
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    5..29
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(99..351)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(371..922)
                     /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(964..1143)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     1460..1480
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        1469..1515
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             2098..2118
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2174..2350
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(2432..2452)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2432..2450)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2471..2487)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2471..2487)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2484..2506)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2495..2511
                     /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(2519..2525,2526..2543,2544..2549))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2564..2585
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2702..2719)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2824..2848
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(2939..3527)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3019..3038)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3618..4406)
                     /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      4864..5299
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 ctggtggcag gatatattgt ggtgtaaaca aattgacgct tagacaactt aataacacat
       61 tgcggacgtt tttaatgtaa tcagcttgca tgccggtcga tctagtaaca tagatgacac
      121 cgcgcgcgat aatttatcct agtttgcgcg ctatattttg ttttctatcg cgtattaaat
      181 gtataattgc gggactctaa tcataaaaac ccatctcata aataacgtca tgcattacat
      241 gttaattatt acatgcttaa cgtaattcaa cagaaattat atgataatca tcgcaagacc
      301 ggcaacagga ttcaatctta agaaacttta ttgccaaatg tttgaacgat ctgcttgact
      361 ctaggggtca tcagatttcg gtgacgggca ggaccggacg gggcggcacc ggcaggctga
      421 agtccagctg ccagaaaccc acgtcatgcc agttcccgtg cttgaagccg gccgcccgca
      481 gcatgccgcg gggggcatat ccgagcgcct cgtgcatgcg cacgctcggg tcgttgggca
      541 gcccgatgac agcgaccacg ctcttgaagc cctgtgcctc cagggacttc agcaggtggg
      601 tgtagagcgt ggagcccagt cccgtccgct ggtggcgggg ggacacgtac acggtcgact
      661 cggccgtcca gtcgtaggcg ttgcgtgcct tccagggacc cgcgtaggcg atgccggcga
      721 cctcgccgtc cacctcggcg acgagccagg gatagcgctc ccgcagacgg acgaggtcgt
      781 ccgtccactc ctgcggttcc tgcggctcgg tacggaagtt gaccgtgctt gtctcgatgt
      841 agtggttgac gatggtgcag accgccggca tgtccgcctc ggtggcacgg cggatgtcgg
      901 ccgggcgtcg ttctgggctc atggtagatc ccctcgatcg agttgagagt gaatatgaga
      961 ctctaattgg ataccgaggg gaatttatgg aacgtcagtg gagcattttt gacaagaaat
     1021 atttgctagc tgatagtgac cttaggcgac ttttgaacgc gcaataatgg tttctgacgt
     1081 atgtgcttag ctcattaaac tccagaaacc cgcggctcag tggctccttc aacgttgcgg
     1141 ttctgtcagt tccaaacgta aaacggcttg tcccgcgtca tcggcggggg tcataacgtg
     1201 actcccttaa ttctccgctc atgatcgata acatgtcgac tatcggacga gctcaaagtt
     1261 taacgtgtga agacgtgctt ctaatgtagt ttaaactgaa ggcgggaaac gacaatctga
     1321 tccaagctca agctaagctt gcatgcctgc aggatatcgt ggatccaagc ttgccacgtg
     1381 ccgccacgtg ccgccacgtg ccgccacgtg cctctagagg atccatctcc actgacgtaa
     1441 gggatgacgc acaatcccac tatccttcgc aagacccttc ctctatataa ggaagttcat
     1501 ttcatttgga gaggacacgc tgggatcccc accatgaacc tgaaggacaa gatcctgggc
     1561 gtggccaaag agctgttcat caagaacggc tacaacgcca ccaccaccgg cgagatcgtg
     1621 aagctgagcg agagcagcaa gggcaacctg tactaccact tcaagaccaa agagaacctg
     1681 ttcctcgaga tcctgaacat cgaggaatcc aagtggcagg aacagtggaa aaaagaacag
     1741 atcaagtgca agacaaaccg cgagaagttc tacctgtaca acgagctgag cctgaccacc
     1801 gagtactact accccctgca gaacgccatc atcgagttct acacagagta ctataagacc
     1861 aacagcatca acgagaagat gaacaagctc gagaacaagt acatcgacgc ctaccacgtg
     1921 atcttcaaag agggcaatct gaacggcgag tggtgcatca atgacgtgaa cgccgtgtcc
     1981 aagatcgccg ccaatgccgt gaatggcatc gtgaccttca cccacgagca gaacattaac
     2041 gagcggatca agctcatgaa caaattcagc cagatcttcc tgaacgggct gagcaagccg
     2101 aagaagaaga ggaaggttta gtgtcggtac gctgaaatca ccagtctctc tctacaaatc
     2161 tatctctctc tattttctcc ataaataatg tgtgagtagt ttcccgataa gggaaattag
     2221 ggttcttata gggtttcgct catgtgttga gcatataaga aacccttagt atgtatttgt
     2281 atttgtaaaa tacttctatc aataaaattt ctaattccta aaaccaaaat ccagtactaa
     2341 aatccagatc gatgcttgat actacgtctt cgggctagag cggactagtt ctagagcggc
     2401 cgccaccgcg gtggagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
     2461 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     2521 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     2581 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     2641 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     2701 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2761 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgaagg
     2821 ccttgacagg atatattggc gggtaaacta agtcgctgta tgtgtttgtt tgagatctca
     2881 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2941 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3001 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3061 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3121 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3181 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3241 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3301 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3361 actacggcta cactagaaga acagtatttg gtatctgtgc tctgctgaag ccagttacct
     3421 tcggaagaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3481 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3541 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3601 tgaattcttg ttagacatta tttgccgact accttggtga tctcgccttt cacgtagtga
     3661 acaaattctt ccaactgatc tgcgcgcgag gccaagcgat cttcttgtcc aagataagcc
     3721 tgcctagctt caagtatgac gggctgatac tgggccggca ggcgctccat tgcccagtcg
     3781 gcagcgacat ccttcggcgc gattttgccg gttactgcgc tgtaccaaat gcgggacaac
     3841 gtaagcacta catttcgctc atcgccagcc cagtcgggcg gcgagttcca tagcgttaag
     3901 gtttcattta gcgcctcaaa tagatcctgt tcaggaaccg gatcaaagag ttcctccgcc
     3961 gctggaccta ccaaggcaac gctatgttct cttgcttttg tcagcaagat agccagatca
     4021 atgtcgatcg tggctggctc gaagatacct gcaagaatgt cattgcgctg ccattctcca
     4081 aattgcagtt cgcgcttagc tggataacgc cacggaatga tgtcgtcgtg cacaacaatg
     4141 gtgacttcta cagcgcggag aatctcgctc tctccagggg aagccgaagt ttccaaaagg
     4201 tcgttgatca aagctcgccg cgttgtttca tcaagcctta cggtcaccgt aaccagcaaa
     4261 tcaatatcac tgtgtggctt caggccgcca tccactgcgg agccgtacaa atgtacggcc
     4321 agcaacgtcg gttcgagatg gcgctcgatg acgccaacta cctctgatag ttgagtcgat
     4381 acttcggcga tcaccgcttc cctcatgatg tttaactcct gaattaagcc gcgccgcgaa
     4441 gcggtgtcgg cttgaatgaa ttgttaggcg tcatcctgtg ctcccgagaa ccagtaccag
     4501 tacatcgctg tttcgttcga gacttgaggt ctagttttat acgtgaacag gtcaatgccg
     4561 ccgagagtaa agccacattt tgcgtacaaa ttgcaggcag gtacattgtt cgtttgtgcc
     4621 atgatgatat atttttatct tgtgcaatgt aacatcagag attttgagac acaacgtggc
     4681 tttgttgaat aaatcgaact tttgctgagt tgaaggatca gatcacgcat cttcccgaca
     4741 acgcagaccg ttccgtggca aagcaaaagt tcaaaatcac caactggtcc acctacaaca
     4801 aagctctcat caaccgtggc tccctcactt tctggctgga tgatggggcg attcaggaga
     4861 tccccatcca acagcccgcc gtcgagcggg cttttttatc cccggaagcc tgtggataga
     4921 gggtagttat ccacgtgaaa ccgctaatgc cccgcaaagc cttgattcac ggggctttcc
     4981 ggcccgctcc aaaaactatc cacgtgaaat cgctaatcag ggtacgtgaa atcgctaatc
     5041 ggagtacgtg aaatcgctaa taaggtcacg tgaaatcgct aatcaaaaag gcacgtgaga
     5101 acgctaatag ccctttcaga tcaacagctt gcaaacaccc ctcgctccgg caagtagtta
     5161 cagcaagtag tatgttcaat tagcttttca attatgaata tatatatcaa ttattggtcg
     5221 cccttggctt gtggacaatg cgctacgcgc accggctccg cccgtggaca accgcaagcg
     5281 gttgcccacc gtcgagcgcc tttgcccaca acccggcggc cggccgcaac agatcgtttt
     5341 ataaattttt tttttttgaa aaagaaaaag cccgaaaggc ggcaacctct cgggcttctg
     5401 gatttccgat ccccggaatt agactctggc tgcctgtatc gagtggtgat tttgtgccga
     5461 gctgccggtc ggggagctgt tggctgg
//
