LOCUS       sequence-387071-        5777 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proG1090::TetR-VP16-19St).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5777)
  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 5777)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: JAB2180
FEATURES             Location/Qualifiers
     source          1..5777
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    29..53
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(123..375)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(395..946)
                     /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(988..1167)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     1484..1504
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        1493..1539
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             2179..2412
                     /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             2413..2433
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2489..2665
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(2747..2767)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2747..2765)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2786..2802)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2786..2802)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2799..2821)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2810..2826
                     /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(2834..2840,2841..2858,2859..2864))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2879..2900
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3017..3034)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3139..3163
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(3254..3842)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3334..3353)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3933..4721)
                     /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      5179..5614
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 gccggtcggg gagctgttgg ctggctggtg gcaggatata ttgtggtgta aacaaattga
       61 cgcttagaca acttaataac acattgcgga cgtttttaat gtaatcagct tgcatgccgg
      121 tcgatctagt aacatagatg acaccgcgcg cgataattta tcctagtttg cgcgctatat
      181 tttgttttct atcgcgtatt aaatgtataa ttgcgggact ctaatcataa aaacccatct
      241 cataaataac gtcatgcatt acatgttaat tattacatgc ttaacgtaat tcaacagaaa
      301 ttatatgata atcatcgcaa gaccggcaac aggattcaat cttaagaaac tttattgcca
      361 aatgtttgaa cgatctgctt gactctaggg gtcatcagat ttcggtgacg ggcaggaccg
      421 gacggggcgg caccggcagg ctgaagtcca gctgccagaa acccacgtca tgccagttcc
      481 cgtgcttgaa gccggccgcc cgcagcatgc cgcggggggc atatccgagc gcctcgtgca
      541 tgcgcacgct cgggtcgttg ggcagcccga tgacagcgac cacgctcttg aagccctgtg
      601 cctccaggga cttcagcagg tgggtgtaga gcgtggagcc cagtcccgtc cgctggtggc
      661 ggggggacac gtacacggtc gactcggccg tccagtcgta ggcgttgcgt gccttccagg
      721 gacccgcgta ggcgatgccg gcgacctcgc cgtccacctc ggcgacgagc cagggatagc
      781 gctcccgcag acggacgagg tcgtccgtcc actcctgcgg ttcctgcggc tcggtacgga
      841 agttgaccgt gcttgtctcg atgtagtggt tgacgatggt gcagaccgcc ggcatgtccg
      901 cctcggtggc acggcggatg tcggccgggc gtcgttctgg gctcatggta gatcccctcg
      961 atcgagttga gagtgaatat gagactctaa ttggataccg aggggaattt atggaacgtc
     1021 agtggagcat ttttgacaag aaatatttgc tagctgatag tgaccttagg cgacttttga
     1081 acgcgcaata atggtttctg acgtatgtgc ttagctcatt aaactccaga aacccgcggc
     1141 tcagtggctc cttcaacgtt gcggttctgt cagttccaaa cgtaaaacgg cttgtcccgc
     1201 gtcatcggcg ggggtcataa cgtgactccc ttaattctcc gctcatgatc gataacatgt
     1261 cgactatcgg acgagctcaa agtttaacgt gtgaagacgt gcttctaatg tagtttaaac
     1321 tgaaggcggg aaacgacaat ctgatccaag ctcaagctaa gcttgcatgc ctgcaggata
     1381 tcgtggatcc aagcttgcca cgtgccgcca cgtgccgcca cgtgccgcca cgtgcctcta
     1441 gaggatccat ctccactgac gtaagggatg acgcacaatc ccactatcct tcgcaagacc
     1501 cttcctctat ataaggaagt tcatttcatt tggagaggac acgctgggat ccccaccatg
     1561 agccggctgg acaagagcaa agtgatcaac agcgccctgg aactgctcaa cgaagtgggc
     1621 atcgagggcc tgaccacccg gaagctggct cagaaactgg gcgtggaaca gcccaccctg
     1681 tactggcacg tgaagaacaa gagagccctg ctggacgccc tggccatcga gatgctggac
     1741 cggcaccaca cccacttttg ccccctggaa ggcgagagct ggcaggattt cctgcggaac
     1801 aacgccaaga gcttcagatg cgctctgctg tcccaccggg atggcgccaa agtgcacctg
     1861 ggcaccagac ctaccgagaa gcagtacgag acactggaaa accagctggc cttcctgtgc
     1921 cagcagggct tcagcctgga aaatgccctg tacgccctga gcgccgtggg ccactttaca
     1981 ctgggctgcg tgctggaaga tcaggaacac caggtcgcca aagaggaaag agagacaccc
     2041 accaccgaca gcatgccccc tctgctgaga caggccattg agctgttcga tcatcagggc
     2101 gccgagcccg cctttctgtt cggcctcgag ctgatcatct gtggcctcga aaaacagctg
     2161 aagtgcgagt ccggcagcgc ccccccgacc gacgtcagcc tgggggacga gctccactta
     2221 gacggcgagg acgtggcgat ggcgcatgcc gacgcgctag acgatttcga tctggacatg
     2281 ttgggggacg gggattcccc ggggccggga tttacccccc acgactccgc cccctacggc
     2341 gctctggata cggccgactt cgagtttgag cagatgttta ccgatgccct tggaattgac
     2401 gagtacggtg ggccgaagaa gaagaggaag gtttagtgtc ggtacgctga aatcaccagt
     2461 ctctctctac aaatctatct ctctctattt tctccataaa taatgtgtga gtagtttccc
     2521 gataagggaa attagggttc ttatagggtt tcgctcatgt gttgagcata taagaaaccc
     2581 ttagtatgta tttgtatttg taaaatactt ctatcaataa aatttctaat tcctaaaacc
     2641 aaaatccagt actaaaatcc agatcgatgc ttgatactac gtcttcgggc tagagcggac
     2701 tagttctaga gcggccgcca ccgcggtgga gctccagctt ttgttccctt tagtgagggt
     2761 taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     2821 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     2881 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     2941 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3001 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3061 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3121 gaaagaacat gaaggccttg acaggatata ttggcgggta aactaagtcg ctgtatgtgt
     3181 ttgtttgaga tctcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3241 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3301 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3361 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3421 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3481 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3541 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3601 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3661 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgtgctctgc
     3721 tgaagccagt taccttcgga agaagagttg gtagctcttg atccggcaaa caaaccaccg
     3781 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3841 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3901 aagggatttt ggtcatgaat tcttgttaga cattatttgc cgactacctt ggtgatctcg
     3961 cctttcacgt agtgaacaaa ttcttccaac tgatctgcgc gcgaggccaa gcgatcttct
     4021 tgtccaagat aagcctgcct agcttcaagt atgacgggct gatactgggc cggcaggcgc
     4081 tccattgccc agtcggcagc gacatccttc ggcgcgattt tgccggttac tgcgctgtac
     4141 caaatgcggg acaacgtaag cactacattt cgctcatcgc cagcccagtc gggcggcgag
     4201 ttccatagcg ttaaggtttc atttagcgcc tcaaatagat cctgttcagg aaccggatca
     4261 aagagttcct ccgccgctgg acctaccaag gcaacgctat gttctcttgc ttttgtcagc
     4321 aagatagcca gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag aatgtcattg
     4381 cgctgccatt ctccaaattg cagttcgcgc ttagctggat aacgccacgg aatgatgtcg
     4441 tcgtgcacaa caatggtgac ttctacagcg cggagaatct cgctctctcc aggggaagcc
     4501 gaagtttcca aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag ccttacggtc
     4561 accgtaacca gcaaatcaat atcactgtgt ggcttcaggc cgccatccac tgcggagccg
     4621 tacaaatgta cggccagcaa cgtcggttcg agatggcgct cgatgacgcc aactacctct
     4681 gatagttgag tcgatacttc ggcgatcacc gcttccctca tgatgtttaa ctcctgaatt
     4741 aagccgcgcc gcgaagcggt gtcggcttga atgaattgtt aggcgtcatc ctgtgctccc
     4801 gagaaccagt accagtacat cgctgtttcg ttcgagactt gaggtctagt tttatacgtg
     4861 aacaggtcaa tgccgccgag agtaaagcca cattttgcgt acaaattgca ggcaggtaca
     4921 ttgttcgttt gtgccatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt
     4981 gagacacaac gtggctttgt tgaataaatc gaacttttgc tgagttgaag gatcagatca
     5041 cgcatcttcc cgacaacgca gaccgttccg tggcaaagca aaagttcaaa atcaccaact
     5101 ggtccaccta caacaaagct ctcatcaacc gtggctccct cactttctgg ctggatgatg
     5161 gggcgattca ggagatcccc atccaacagc ccgccgtcga gcgggctttt ttatccccgg
     5221 aagcctgtgg atagagggta gttatccacg tgaaaccgct aatgccccgc aaagccttga
     5281 ttcacggggc tttccggccc gctccaaaaa ctatccacgt gaaatcgcta atcagggtac
     5341 gtgaaatcgc taatcggagt acgtgaaatc gctaataagg tcacgtgaaa tcgctaatca
     5401 aaaaggcacg tgagaacgct aatagccctt tcagatcaac agcttgcaaa cacccctcgc
     5461 tccggcaagt agttacagca agtagtatgt tcaattagct tttcaattat gaatatatat
     5521 atcaattatt ggtcgccctt ggcttgtgga caatgcgcta cgcgcaccgg ctccgcccgt
     5581 ggacaaccgc aagcggttgc ccaccgtcga gcgcctttgc ccacaacccg gcggccggcc
     5641 gcaacagatc gttttataaa tttttttttt tgaaaaagaa aaagcccgaa aggcggcaac
     5701 ctctcgggct tctggatttc cgatccccgg aattagactc tggctgcctg tatcgagtgg
     5761 tgattttgtg ccgagct
//
