LOCUS       sequence-387093-        5894 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proG1090::RpaR-VP16_19St).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5894)
  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 5894)
  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: JAB2494
FEATURES             Location/Qualifiers
     source          1..5894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(88..340)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(360..911)
                     /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(953..1132)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     1449..1469
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        1458..1504
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             2261..2494
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDTADFEFEQMFTDALGIDEYGG"
     CDS             2495..2515
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2571..2747
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(2829..2849)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2829..2847)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2868..2884)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2868..2884)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2881..2903)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2892..2908
                     /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(2916..2922,2923..2940,2941..2946))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2961..2982
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3099..3116)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3221..3245
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(3336..3924)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3416..3435)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4015..4803)
                     /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      5261..5696
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(5888..5894,1..18)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 atatattgtg gtgtaaacaa attgacgctt agacaactta ataacacatt gcggacgttt
       61 ttaatgtaat cagcttgcat gccggtcgat ctagtaacat agatgacacc gcgcgcgata
      121 atttatccta gtttgcgcgc tatattttgt tttctatcgc gtattaaatg tataattgcg
      181 ggactctaat cataaaaacc catctcataa ataacgtcat gcattacatg ttaattatta
      241 catgcttaac gtaattcaac agaaattata tgataatcat cgcaagaccg gcaacaggat
      301 tcaatcttaa gaaactttat tgccaaatgt ttgaacgatc tgcttgactc taggggtcat
      361 cagatttcgg tgacgggcag gaccggacgg ggcggcaccg gcaggctgaa gtccagctgc
      421 cagaaaccca cgtcatgcca gttcccgtgc ttgaagccgg ccgcccgcag catgccgcgg
      481 ggggcatatc cgagcgcctc gtgcatgcgc acgctcgggt cgttgggcag cccgatgaca
      541 gcgaccacgc tcttgaagcc ctgtgcctcc agggacttca gcaggtgggt gtagagcgtg
      601 gagcccagtc ccgtccgctg gtggcggggg gacacgtaca cggtcgactc ggccgtccag
      661 tcgtaggcgt tgcgtgcctt ccagggaccc gcgtaggcga tgccggcgac ctcgccgtcc
      721 acctcggcga cgagccaggg atagcgctcc cgcagacgga cgaggtcgtc cgtccactcc
      781 tgcggttcct gcggctcggt acggaagttg accgtgcttg tctcgatgta gtggttgacg
      841 atggtgcaga ccgccggcat gtccgcctcg gtggcacggc ggatgtcggc cgggcgtcgt
      901 tctgggctca tggtagatcc cctcgatcga gttgagagtg aatatgagac tctaattgga
      961 taccgagggg aatttatgga acgtcagtgg agcatttttg acaagaaata tttgctagct
     1021 gatagtgacc ttaggcgact tttgaacgcg caataatggt ttctgacgta tgtgcttagc
     1081 tcattaaact ccagaaaccc gcggctcagt ggctccttca acgttgcggt tctgtcagtt
     1141 ccaaacgtaa aacggcttgt cccgcgtcat cggcgggggt cataacgtga ctcccttaat
     1201 tctccgctca tgatcgataa catgtcgact atcggacgag ctcaaagttt aacgtgtgaa
     1261 gacgtgcttc taatgtagtt taaactgaag gcgggaaacg acaatctgat ccaagctcaa
     1321 gctaagcttg catgcctgca ggatatcgtg gatccaagct tgccacgtgc cgccacgtgc
     1381 cgccacgtgc cgccacgtgc ctctagagga tccatctcca ctgacgtaag ggatgacgca
     1441 caatcccact atccttcgca agacccttcc tctatataag gaagttcatt tcatttggag
     1501 aggacacgct gggatcccca ccatgattgt gggtgaagat cagctgtggg gtcgtcgtac
     1561 actggaattt gttgatagcg ttgaacgtct ggaagcaccg gcactgatta gccgttttga
     1621 aagcctgatt gcaagctgtg gttttaccgc ctatatcatg gcaggtctgc cgagccgtaa
     1681 tgccggtctg ccggaactga ccctggcaaa tggttggcct cgtgattggt ttgatctgta
     1741 tgttagcgaa aactttagcg cagttgatcc ggttccgcgt tatggtgcaa ccactgttca
     1801 tccgtttgtt tggagtgatg caccgtatga tcgtgaccgt gatcaggcag cacatcgtgt
     1861 tatgacccgt gcagcagaat ttggtctggt tgaaggttat tgtattccgc tgcattacga
     1921 tgatggtagc gcagcaatta gtatggcagg tgaagatcct gatctgagtc cggcagcccg
     1981 tggtgtaatg cagctggtta gcatttatgc acatagccgt ctgcgtgtac tgagccgtcc
     2041 gaaaccgatt cgtcgtaatc gtctgacacc gcgtgaatgt gaaattctgc agtgggcagc
     2101 acagggtaaa accgcatggg aaattagcgt tattctgtgt attaccgaac gcaccgttaa
     2161 atttcatctg attgaagcag cacgtaaact ggatgcagca aatcgtaccg cagcagttgc
     2221 aaaagcactg acactgggtc tgattcgtct gggaggaggt gcccccccga ccgacgtcag
     2281 cctgggggac gagctccact tagacggcga ggacgtggcg atggcgcatg ccgacgcgct
     2341 agacgatttc gatctggaca tgttggggga cggggattcc ccggggccgg gatttacccc
     2401 ccacgactcc gccccctacg gcgctctgga tacggccgac ttcgagtttg agcagatgtt
     2461 taccgatgcc cttggaattg acgagtacgg tgggccgaag aagaagagga aggtttagtg
     2521 tcggtacgct gaaatcacca gtctctctct acaaatctat ctctctctat tttctccata
     2581 aataatgtgt gagtagtttc ccgataaggg aaattagggt tcttataggg tttcgctcat
     2641 gtgttgagca tataagaaac ccttagtatg tatttgtatt tgtaaaatac ttctatcaat
     2701 aaaatttcta attcctaaaa ccaaaatcca gtactaaaat ccagatcgat gcttgatact
     2761 acgtcttcgg gctagagcgg actagttcta gagcggccgc caccgcggtg gagctccagc
     2821 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
     2881 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     2941 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     3001 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     3061 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3121 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3181 acagaatcag gggataacgc aggaaagaac atgaaggcct tgacaggata tattggcggg
     3241 taaactaagt cgctgtatgt gtttgtttga gatctcatgt gagcaaaagg ccagcaaaag
     3301 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3361 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3421 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3481 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3541 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3601 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3661 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3721 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3781 gtatttggta tctgtgctct gctgaagcca gttaccttcg gaagaagagt tggtagctct
     3841 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3901 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3961 cagtggaacg aaaactcacg ttaagggatt ttggtcatga attcttgtta gacattattt
     4021 gccgactacc ttggtgatct cgcctttcac gtagtgaaca aattcttcca actgatctgc
     4081 gcgcgaggcc aagcgatctt cttgtccaag ataagcctgc ctagcttcaa gtatgacggg
     4141 ctgatactgg gccggcaggc gctccattgc ccagtcggca gcgacatcct tcggcgcgat
     4201 tttgccggtt actgcgctgt accaaatgcg ggacaacgta agcactacat ttcgctcatc
     4261 gccagcccag tcgggcggcg agttccatag cgttaaggtt tcatttagcg cctcaaatag
     4321 atcctgttca ggaaccggat caaagagttc ctccgccgct ggacctacca aggcaacgct
     4381 atgttctctt gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa
     4441 gatacctgca agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg
     4501 ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg acttctacag cgcggagaat
     4561 ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt
     4621 tgtttcatca agccttacgg tcaccgtaac cagcaaatca atatcactgt gtggcttcag
     4681 gccgccatcc actgcggagc cgtacaaatg tacggccagc aacgtcggtt cgagatggcg
     4741 ctcgatgacg ccaactacct ctgatagttg agtcgatact tcggcgatca ccgcttccct
     4801 catgatgttt aactcctgaa ttaagccgcg ccgcgaagcg gtgtcggctt gaatgaattg
     4861 ttaggcgtca tcctgtgctc ccgagaacca gtaccagtac atcgctgttt cgttcgagac
     4921 ttgaggtcta gttttatacg tgaacaggtc aatgccgccg agagtaaagc cacattttgc
     4981 gtacaaattg caggcaggta cattgttcgt ttgtgccatg atgatatatt tttatcttgt
     5041 gcaatgtaac atcagagatt ttgagacaca acgtggcttt gttgaataaa tcgaactttt
     5101 gctgagttga aggatcagat cacgcatctt cccgacaacg cagaccgttc cgtggcaaag
     5161 caaaagttca aaatcaccaa ctggtccacc tacaacaaag ctctcatcaa ccgtggctcc
     5221 ctcactttct ggctggatga tggggcgatt caggagatcc ccatccaaca gcccgccgtc
     5281 gagcgggctt ttttatcccc ggaagcctgt ggatagaggg tagttatcca cgtgaaaccg
     5341 ctaatgcccc gcaaagcctt gattcacggg gctttccggc ccgctccaaa aactatccac
     5401 gtgaaatcgc taatcagggt acgtgaaatc gctaatcgga gtacgtgaaa tcgctaataa
     5461 ggtcacgtga aatcgctaat caaaaaggca cgtgagaacg ctaatagccc tttcagatca
     5521 acagcttgca aacacccctc gctccggcaa gtagttacag caagtagtat gttcaattag
     5581 cttttcaatt atgaatatat atatcaatta ttggtcgccc ttggcttgtg gacaatgcgc
     5641 tacgcgcacc ggctccgccc gtggacaacc gcaagcggtt gcccaccgtc gagcgccttt
     5701 gcccacaacc cggcggccgg ccgcaacaga tcgttttata aatttttttt tttgaaaaag
     5761 aaaaagcccg aaaggcggca acctctcggg cttctggatt tccgatcccc ggaattagac
     5821 tctggctgcc tgtatcgagt ggtgattttg tgccgagctg ccggtcgggg agctgttggc
     5881 tggctggtgg cagg
//
