LOCUS       sequence-393420-        5588 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proG1090::AmtR-NLS_19St).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5588)
  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 5588)
  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: JAB2487
FEATURES             Location/Qualifiers
     source          1..5588
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    7..31
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(101..353)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(373..924)
                     /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(966..1145)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     1462..1482
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        1471..1517
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             2202..2222
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2278..2454
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(2536..2556)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2536..2554)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2575..2591)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2575..2591)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2588..2610)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2599..2615
                     /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(2623..2629,2630..2647,2648..2653))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2668..2689
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2806..2823)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2928..2952
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(3043..3631)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3123..3142)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3722..4510)
                     /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      4968..5403
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 ggctggtggc aggatatatt gtggtgtaaa caaattgacg cttagacaac ttaataacac
       61 attgcggacg tttttaatgt aatcagcttg catgccggtc gatctagtaa catagatgac
      121 accgcgcgcg ataatttatc ctagtttgcg cgctatattt tgttttctat cgcgtattaa
      181 atgtataatt gcgggactct aatcataaaa acccatctca taaataacgt catgcattac
      241 atgttaatta ttacatgctt aacgtaattc aacagaaatt atatgataat catcgcaaga
      301 ccggcaacag gattcaatct taagaaactt tattgccaaa tgtttgaacg atctgcttga
      361 ctctaggggt catcagattt cggtgacggg caggaccgga cggggcggca ccggcaggct
      421 gaagtccagc tgccagaaac ccacgtcatg ccagttcccg tgcttgaagc cggccgcccg
      481 cagcatgccg cggggggcat atccgagcgc ctcgtgcatg cgcacgctcg ggtcgttggg
      541 cagcccgatg acagcgacca cgctcttgaa gccctgtgcc tccagggact tcagcaggtg
      601 ggtgtagagc gtggagccca gtcccgtccg ctggtggcgg ggggacacgt acacggtcga
      661 ctcggccgtc cagtcgtagg cgttgcgtgc cttccaggga cccgcgtagg cgatgccggc
      721 gacctcgccg tccacctcgg cgacgagcca gggatagcgc tcccgcagac ggacgaggtc
      781 gtccgtccac tcctgcggtt cctgcggctc ggtacggaag ttgaccgtgc ttgtctcgat
      841 gtagtggttg acgatggtgc agaccgccgg catgtccgcc tcggtggcac ggcggatgtc
      901 ggccgggcgt cgttctgggc tcatggtaga tcccctcgat cgagttgaga gtgaatatga
      961 gactctaatt ggataccgag gggaatttat ggaacgtcag tggagcattt ttgacaagaa
     1021 atatttgcta gctgatagtg accttaggcg acttttgaac gcgcaataat ggtttctgac
     1081 gtatgtgctt agctcattaa actccagaaa cccgcggctc agtggctcct tcaacgttgc
     1141 ggttctgtca gttccaaacg taaaacggct tgtcccgcgt catcggcggg ggtcataacg
     1201 tgactccctt aattctccgc tcatgatcga taacatgtcg actatcggac gagctcaaag
     1261 tttaacgtgt gaagacgtgc ttctaatgta gtttaaactg aaggcgggaa acgacaatct
     1321 gatccaagct caagctaagc ttgcatgcct gcaggatatc gtggatccaa gcttgccacg
     1381 tgccgccacg tgccgccacg tgccgccacg tgcctctaga ggatccatct ccactgacgt
     1441 aagggatgac gcacaatccc actatccttc gcaagaccct tcctctatat aaggaagttc
     1501 atttcatttg gagaggacac gctgggatcc ccaccatggc tggcgccgtg ggcagaccca
     1561 gaagatctgc tcctcggaga gccggcaaga acccccggga agagattctg gatgccagcg
     1621 ccgagctgtt caccagacag ggctttgcca ccaccagcac ccaccagatt gccgacgctg
     1681 tgggcatcag acaggccagc ctgtactacc acttccccag caagaccgag atcttcctga
     1741 ccctgctgaa aagcaccgtg gaaccctcca ccgtgctggc cgaggatctg tctaccctgg
     1801 acgccggacc cgaaatgaga ctgtgggcta tcgtggccag cgaagtgcgg ctgctgctga
     1861 gcaccaagtg gaacgtgggc cggctgtacc agctgcccat cgtgggctct gaggaattcg
     1921 ccgagtacca cagccagcgc gaggccctga ccaacgtgtt cagagatctg gccaccgaga
     1981 ttgtgggcga cgaccccaga gccgaactgc ccttccacat caccatgagc gtgatcgaga
     2041 tgcggcggaa cgacggcaag atccctagcc ctctgagcgc cgactctctg cccgagacag
     2101 ccattatgct ggctgacgcc tccctggctg tgctgggagc acctctgcct gccgacagag
     2161 tggaaaagac actggaactg atcaagcagg ccgacgccaa gccgaagaag aagaggaagg
     2221 tttagtgtcg gtacgctgaa atcaccagtc tctctctaca aatctatctc tctctatttt
     2281 ctccataaat aatgtgtgag tagtttcccg ataagggaaa ttagggttct tatagggttt
     2341 cgctcatgtg ttgagcatat aagaaaccct tagtatgtat ttgtatttgt aaaatacttc
     2401 tatcaataaa atttctaatt cctaaaacca aaatccagta ctaaaatcca gatcgatgct
     2461 tgatactacg tcttcgggct agagcggact agttctagag cggccgccac cgcggtggag
     2521 ctccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
     2581 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     2641 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     2701 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     2761 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2821 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2881 gttatccaca gaatcagggg ataacgcagg aaagaacatg aaggccttga caggatatat
     2941 tggcgggtaa actaagtcgc tgtatgtgtt tgtttgagat ctcatgtgag caaaaggcca
     3001 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3061 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3121 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3181 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3241 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3301 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3361 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3421 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3481 aagaacagta tttggtatct gtgctctgct gaagccagtt accttcggaa gaagagttgg
     3541 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3601 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3661 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgaatt cttgttagac
     3721 attatttgcc gactaccttg gtgatctcgc ctttcacgta gtgaacaaat tcttccaact
     3781 gatctgcgcg cgaggccaag cgatcttctt gtccaagata agcctgccta gcttcaagta
     3841 tgacgggctg atactgggcc ggcaggcgct ccattgccca gtcggcagcg acatccttcg
     3901 gcgcgatttt gccggttact gcgctgtacc aaatgcggga caacgtaagc actacatttc
     3961 gctcatcgcc agcccagtcg ggcggcgagt tccatagcgt taaggtttca tttagcgcct
     4021 caaatagatc ctgttcagga accggatcaa agagttcctc cgccgctgga cctaccaagg
     4081 caacgctatg ttctcttgct tttgtcagca agatagccag atcaatgtcg atcgtggctg
     4141 gctcgaagat acctgcaaga atgtcattgc gctgccattc tccaaattgc agttcgcgct
     4201 tagctggata acgccacgga atgatgtcgt cgtgcacaac aatggtgact tctacagcgc
     4261 ggagaatctc gctctctcca ggggaagccg aagtttccaa aaggtcgttg atcaaagctc
     4321 gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag caaatcaata tcactgtgtg
     4381 gcttcaggcc gccatccact gcggagccgt acaaatgtac ggccagcaac gtcggttcga
     4441 gatggcgctc gatgacgcca actacctctg atagttgagt cgatacttcg gcgatcaccg
     4501 cttccctcat gatgtttaac tcctgaatta agccgcgccg cgaagcggtg tcggcttgaa
     4561 tgaattgtta ggcgtcatcc tgtgctcccg agaaccagta ccagtacatc gctgtttcgt
     4621 tcgagacttg aggtctagtt ttatacgtga acaggtcaat gccgccgaga gtaaagccac
     4681 attttgcgta caaattgcag gcaggtacat tgttcgtttg tgccatgatg atatattttt
     4741 atcttgtgca atgtaacatc agagattttg agacacaacg tggctttgtt gaataaatcg
     4801 aacttttgct gagttgaagg atcagatcac gcatcttccc gacaacgcag accgttccgt
     4861 ggcaaagcaa aagttcaaaa tcaccaactg gtccacctac aacaaagctc tcatcaaccg
     4921 tggctccctc actttctggc tggatgatgg ggcgattcag gagatcccca tccaacagcc
     4981 cgccgtcgag cgggcttttt tatccccgga agcctgtgga tagagggtag ttatccacgt
     5041 gaaaccgcta atgccccgca aagccttgat tcacggggct ttccggcccg ctccaaaaac
     5101 tatccacgtg aaatcgctaa tcagggtacg tgaaatcgct aatcggagta cgtgaaatcg
     5161 ctaataaggt cacgtgaaat cgctaatcaa aaaggcacgt gagaacgcta atagcccttt
     5221 cagatcaaca gcttgcaaac acccctcgct ccggcaagta gttacagcaa gtagtatgtt
     5281 caattagctt ttcaattatg aatatatata tcaattattg gtcgcccttg gcttgtggac
     5341 aatgcgctac gcgcaccggc tccgcccgtg gacaaccgca agcggttgcc caccgtcgag
     5401 cgcctttgcc cacaacccgg cggccggccg caacagatcg ttttataaat tttttttttt
     5461 gaaaaagaaa aagcccgaaa ggcggcaacc tctcgggctt ctggatttcc gatccccgga
     5521 attagactct ggctgcctgt atcgagtggt gattttgtgc cgagctgccg gtcggggagc
     5581 tgttggct
//
