LOCUS       sequence-387021-        4463 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (empty vector control).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4463)
  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 4463)
  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: JAB2121
FEATURES             Location/Qualifiers
     source          1..4463
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    7..31
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(122..710)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(202..221)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(801..1589)
                     /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      2047..2482
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    2674..2698
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(2768..3020)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(3040..3591)
                     /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(3633..3812)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     complement(4078..4098)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4078..4096)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4117..4133)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4117..4133)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4130..4152)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4141..4157
                     /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(4165..4171,4172..4189,4190..4195))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4210..4231
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4348..4365)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aggccttgac aggatatatt ggcgggtaaa ctaagtcgct gtatgtgttt gtttgagatc
       61 tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      121 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      181 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      241 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      301 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      361 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      421 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      481 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      541 ctaactacgg ctacactaga agaacagtat ttggtatctg tgctctgctg aagccagtta
      601 ccttcggaag aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      661 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      721 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      781 tcatgaattc ttgttagaca ttatttgccg actaccttgg tgatctcgcc tttcacgtag
      841 tgaacaaatt cttccaactg atctgcgcgc gaggccaagc gatcttcttg tccaagataa
      901 gcctgcctag cttcaagtat gacgggctga tactgggccg gcaggcgctc cattgcccag
      961 tcggcagcga catccttcgg cgcgattttg ccggttactg cgctgtacca aatgcgggac
     1021 aacgtaagca ctacatttcg ctcatcgcca gcccagtcgg gcggcgagtt ccatagcgtt
     1081 aaggtttcat ttagcgcctc aaatagatcc tgttcaggaa ccggatcaaa gagttcctcc
     1141 gccgctggac ctaccaaggc aacgctatgt tctcttgctt ttgtcagcaa gatagccaga
     1201 tcaatgtcga tcgtggctgg ctcgaagata cctgcaagaa tgtcattgcg ctgccattct
     1261 ccaaattgca gttcgcgctt agctggataa cgccacggaa tgatgtcgtc gtgcacaaca
     1321 atggtgactt ctacagcgcg gagaatctcg ctctctccag gggaagccga agtttccaaa
     1381 aggtcgttga tcaaagctcg ccgcgttgtt tcatcaagcc ttacggtcac cgtaaccagc
     1441 aaatcaatat cactgtgtgg cttcaggccg ccatccactg cggagccgta caaatgtacg
     1501 gccagcaacg tcggttcgag atggcgctcg atgacgccaa ctacctctga tagttgagtc
     1561 gatacttcgg cgatcaccgc ttccctcatg atgtttaact cctgaattaa gccgcgccgc
     1621 gaagcggtgt cggcttgaat gaattgttag gcgtcatcct gtgctcccga gaaccagtac
     1681 cagtacatcg ctgtttcgtt cgagacttga ggtctagttt tatacgtgaa caggtcaatg
     1741 ccgccgagag taaagccaca ttttgcgtac aaattgcagg caggtacatt gttcgtttgt
     1801 gccatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga gacacaacgt
     1861 ggctttgttg aataaatcga acttttgctg agttgaagga tcagatcacg catcttcccg
     1921 acaacgcaga ccgttccgtg gcaaagcaaa agttcaaaat caccaactgg tccacctaca
     1981 acaaagctct catcaaccgt ggctccctca ctttctggct ggatgatggg gcgattcagg
     2041 agatccccat ccaacagccc gccgtcgagc gggctttttt atccccggaa gcctgtggat
     2101 agagggtagt tatccacgtg aaaccgctaa tgccccgcaa agccttgatt cacggggctt
     2161 tccggcccgc tccaaaaact atccacgtga aatcgctaat cagggtacgt gaaatcgcta
     2221 atcggagtac gtgaaatcgc taataaggtc acgtgaaatc gctaatcaaa aaggcacgtg
     2281 agaacgctaa tagccctttc agatcaacag cttgcaaaca cccctcgctc cggcaagtag
     2341 ttacagcaag tagtatgttc aattagcttt tcaattatga atatatatat caattattgg
     2401 tcgcccttgg cttgtggaca atgcgctacg cgcaccggct ccgcccgtgg acaaccgcaa
     2461 gcggttgccc accgtcgagc gcctttgccc acaacccggc ggccggccgc aacagatcgt
     2521 tttataaatt tttttttttg aaaaagaaaa agcccgaaag gcggcaacct ctcgggcttc
     2581 tggatttccg atccccggaa ttagactctg gctgcctgta tcgagtggtg attttgtgcc
     2641 gagctgccgg tcggggagct gttggctggc tggtggcagg atatattgtg gtgtaaacaa
     2701 attgacgctt agacaactta ataacacatt gcggacgttt ttaatgtaat cagcttgcat
     2761 gccggtcgat ctagtaacat agatgacacc gcgcgcgata atttatccta gtttgcgcgc
     2821 tatattttgt tttctatcgc gtattaaatg tataattgcg ggactctaat cataaaaacc
     2881 catctcataa ataacgtcat gcattacatg ttaattatta catgcttaac gtaattcaac
     2941 agaaattata tgataatcat cgcaagaccg gcaacaggat tcaatcttaa gaaactttat
     3001 tgccaaatgt ttgaacgatc tgcttgactc taggggtcat cagatttcgg tgacgggcag
     3061 gaccggacgg ggcggcaccg gcaggctgaa gtccagctgc cagaaaccca cgtcatgcca
     3121 gttcccgtgc ttgaagccgg ccgcccgcag catgccgcgg ggggcatatc cgagcgcctc
     3181 gtgcatgcgc acgctcgggt cgttgggcag cccgatgaca gcgaccacgc tcttgaagcc
     3241 ctgtgcctcc agggacttca gcaggtgggt gtagagcgtg gagcccagtc ccgtccgctg
     3301 gtggcggggg gacacgtaca cggtcgactc ggccgtccag tcgtaggcgt tgcgtgcctt
     3361 ccagggaccc gcgtaggcga tgccggcgac ctcgccgtcc acctcggcga cgagccaggg
     3421 atagcgctcc cgcagacgga cgaggtcgtc cgtccactcc tgcggttcct gcggctcggt
     3481 acggaagttg accgtgcttg tctcgatgta gtggttgacg atggtgcaga ccgccggcat
     3541 gtccgcctcg gtggcacggc ggatgtcggc cgggcgtcgt tctgggctca tggtagatcc
     3601 cctcgatcga gttgagagtg aatatgagac tctaattgga taccgagggg aatttatgga
     3661 acgtcagtgg agcatttttg acaagaaata tttgctagct gatagtgacc ttaggcgact
     3721 tttgaacgcg caataatggt ttctgacgta tgtgcttagc tcattaaact ccagaaaccc
     3781 gcggctcagt ggctccttca acgttgcggt tctgtcagtt ccaaacgtaa aacggcttgt
     3841 cccgcgtcat cggcgggggt cataacgtga ctcccttaat tctccgctca tgatcgataa
     3901 catgtcgact atcggacgag ctcaaagttt aacgtgtgaa gacgtgcttc taatgcgaga
     3961 ccacccggcg ctaacgcact caaagccggg acggtctctg cttgatacta cgtcttcggg
     4021 ctagagcgga ctagttctag agcggccgcc accgcggtgg agctccagct tttgttccct
     4081 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4141 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4201 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4261 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4321 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4381 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4441 ggataacgca ggaaagaaca tga
//
