LOCUS       sequence-387076-        5522 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proG1090::PhlF-NLS-19Sterm).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5522)
  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 5522)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: JAB2240
FEATURES             Location/Qualifiers
     source          1..5522
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    9..33
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(103..355)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(375..926)
                     /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(968..1147)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     1464..1484
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        1473..1519
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             2138..2158
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2214..2390
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(2472..2492)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2472..2490)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2511..2527)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2511..2527)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2524..2546)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2535..2551
                     /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(2559..2565,2566..2583,2584..2589))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2604..2625
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2742..2759)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2864..2888
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(2979..3567)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3059..3078)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3658..4446)
                     /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      4904..5339
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 ctggctggtg gcaggatata ttgtggtgta aacaaattga cgcttagaca acttaataac
       61 acattgcgga cgtttttaat gtaatcagct tgcatgccgg tcgatctagt aacatagatg
      121 acaccgcgcg cgataattta tcctagtttg cgcgctatat tttgttttct atcgcgtatt
      181 aaatgtataa ttgcgggact ctaatcataa aaacccatct cataaataac gtcatgcatt
      241 acatgttaat tattacatgc ttaacgtaat tcaacagaaa ttatatgata atcatcgcaa
      301 gaccggcaac aggattcaat cttaagaaac tttattgcca aatgtttgaa cgatctgctt
      361 gactctaggg gtcatcagat ttcggtgacg ggcaggaccg gacggggcgg caccggcagg
      421 ctgaagtcca gctgccagaa acccacgtca tgccagttcc cgtgcttgaa gccggccgcc
      481 cgcagcatgc cgcggggggc atatccgagc gcctcgtgca tgcgcacgct cgggtcgttg
      541 ggcagcccga tgacagcgac cacgctcttg aagccctgtg cctccaggga cttcagcagg
      601 tgggtgtaga gcgtggagcc cagtcccgtc cgctggtggc ggggggacac gtacacggtc
      661 gactcggccg tccagtcgta ggcgttgcgt gccttccagg gacccgcgta ggcgatgccg
      721 gcgacctcgc cgtccacctc ggcgacgagc cagggatagc gctcccgcag acggacgagg
      781 tcgtccgtcc actcctgcgg ttcctgcggc tcggtacgga agttgaccgt gcttgtctcg
      841 atgtagtggt tgacgatggt gcagaccgcc ggcatgtccg cctcggtggc acggcggatg
      901 tcggccgggc gtcgttctgg gctcatggta gatcccctcg atcgagttga gagtgaatat
      961 gagactctaa ttggataccg aggggaattt atggaacgtc agtggagcat ttttgacaag
     1021 aaatatttgc tagctgatag tgaccttagg cgacttttga acgcgcaata atggtttctg
     1081 acgtatgtgc ttagctcatt aaactccaga aacccgcggc tcagtggctc cttcaacgtt
     1141 gcggttctgt cagttccaaa cgtaaaacgg cttgtcccgc gtcatcggcg ggggtcataa
     1201 cgtgactccc ttaattctcc gctcatgatc gataacatgt cgactatcgg acgagctcaa
     1261 agtttaacgt gtgaagacgt gcttctaatg tagtttaaac tgaaggcggg aaacgacaat
     1321 ctgatccaag ctcaagctaa gcttgcatgc ctgcaggata tcgtggatcc aagcttgcca
     1381 cgtgccgcca cgtgccgcca cgtgccgcca cgtgcctcta gaggatccat ctccactgac
     1441 gtaagggatg acgcacaatc ccactatcct tcgcaagacc cttcctctat ataaggaagt
     1501 tcatttcatt tggagaggac acgctgggat ccccaccatg gcccggaccc ccagcagaag
     1561 cagcatcgga tctctgagaa gcccccacac ccacaaggcc atcctgacca gcaccatcga
     1621 gatcctgaaa gagtgcggct acagcggcct gagcatcgag tctgtggcta gaagggctgg
     1681 cgcctccaag cccaccatct accggtggtg gacaaacaag gccgccctga tcgccgaggt
     1741 gtacgagaac gagagcgagc aggtccgaaa gttccccgac ctgggcagct tcaaggccga
     1801 tctggacttc ctgctgcgga acctgtggaa agtgtggcgg gaaaccatct gcggcgaggc
     1861 cttcagatgc gtgatcgctg aggcccagct ggaccctgcc acactgaccc agctgaagga
     1921 ccagttcatg gaacggcgga gagagatgcc caagaaactg gtggaaaacg ccatcagcaa
     1981 cggcgagctg cccaaggaca ccaaccggga actgctgctg gatatgatct tcggcttctg
     2041 ctggtacaga ctgctgaccg agcagctgac cgtggaacag gatatcgagg aattcacctt
     2101 tctgctgatc aacggcgtgt gccccggcac ccagagaccg aagaagaaga ggaaggttta
     2161 gtgtcggtac gctgaaatca ccagtctctc tctacaaatc tatctctctc tattttctcc
     2221 ataaataatg tgtgagtagt ttcccgataa gggaaattag ggttcttata gggtttcgct
     2281 catgtgttga gcatataaga aacccttagt atgtatttgt atttgtaaaa tacttctatc
     2341 aataaaattt ctaattccta aaaccaaaat ccagtactaa aatccagatc gatgcttgat
     2401 actacgtctt cgggctagag cggactagtt ctagagcggc cgccaccgcg gtggagctcc
     2461 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
     2521 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     2581 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     2641 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     2701 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     2761 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2821 tccacagaat caggggataa cgcaggaaag aacatgaagg ccttgacagg atatattggc
     2881 gggtaaacta agtcgctgta tgtgtttgtt tgagatctca tgtgagcaaa aggccagcaa
     2941 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3001 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3061 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3121 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3181 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3241 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3301 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3361 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3421 acagtatttg gtatctgtgc tctgctgaag ccagttacct tcggaagaag agttggtagc
     3481 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3541 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3601 gctcagtgga acgaaaactc acgttaaggg attttggtca tgaattcttg ttagacatta
     3661 tttgccgact accttggtga tctcgccttt cacgtagtga acaaattctt ccaactgatc
     3721 tgcgcgcgag gccaagcgat cttcttgtcc aagataagcc tgcctagctt caagtatgac
     3781 gggctgatac tgggccggca ggcgctccat tgcccagtcg gcagcgacat ccttcggcgc
     3841 gattttgccg gttactgcgc tgtaccaaat gcgggacaac gtaagcacta catttcgctc
     3901 atcgccagcc cagtcgggcg gcgagttcca tagcgttaag gtttcattta gcgcctcaaa
     3961 tagatcctgt tcaggaaccg gatcaaagag ttcctccgcc gctggaccta ccaaggcaac
     4021 gctatgttct cttgcttttg tcagcaagat agccagatca atgtcgatcg tggctggctc
     4081 gaagatacct gcaagaatgt cattgcgctg ccattctcca aattgcagtt cgcgcttagc
     4141 tggataacgc cacggaatga tgtcgtcgtg cacaacaatg gtgacttcta cagcgcggag
     4201 aatctcgctc tctccagggg aagccgaagt ttccaaaagg tcgttgatca aagctcgccg
     4261 cgttgtttca tcaagcctta cggtcaccgt aaccagcaaa tcaatatcac tgtgtggctt
     4321 caggccgcca tccactgcgg agccgtacaa atgtacggcc agcaacgtcg gttcgagatg
     4381 gcgctcgatg acgccaacta cctctgatag ttgagtcgat acttcggcga tcaccgcttc
     4441 cctcatgatg tttaactcct gaattaagcc gcgccgcgaa gcggtgtcgg cttgaatgaa
     4501 ttgttaggcg tcatcctgtg ctcccgagaa ccagtaccag tacatcgctg tttcgttcga
     4561 gacttgaggt ctagttttat acgtgaacag gtcaatgccg ccgagagtaa agccacattt
     4621 tgcgtacaaa ttgcaggcag gtacattgtt cgtttgtgcc atgatgatat atttttatct
     4681 tgtgcaatgt aacatcagag attttgagac acaacgtggc tttgttgaat aaatcgaact
     4741 tttgctgagt tgaaggatca gatcacgcat cttcccgaca acgcagaccg ttccgtggca
     4801 aagcaaaagt tcaaaatcac caactggtcc acctacaaca aagctctcat caaccgtggc
     4861 tccctcactt tctggctgga tgatggggcg attcaggaga tccccatcca acagcccgcc
     4921 gtcgagcggg cttttttatc cccggaagcc tgtggataga gggtagttat ccacgtgaaa
     4981 ccgctaatgc cccgcaaagc cttgattcac ggggctttcc ggcccgctcc aaaaactatc
     5041 cacgtgaaat cgctaatcag ggtacgtgaa atcgctaatc ggagtacgtg aaatcgctaa
     5101 taaggtcacg tgaaatcgct aatcaaaaag gcacgtgaga acgctaatag ccctttcaga
     5161 tcaacagctt gcaaacaccc ctcgctccgg caagtagtta cagcaagtag tatgttcaat
     5221 tagcttttca attatgaata tatatatcaa ttattggtcg cccttggctt gtggacaatg
     5281 cgctacgcgc accggctccg cccgtggaca accgcaagcg gttgcccacc gtcgagcgcc
     5341 tttgcccaca acccggcggc cggccgcaac agatcgtttt ataaattttt ttttttgaaa
     5401 aagaaaaagc ccgaaaggcg gcaacctctc gggcttctgg atttccgatc cccggaatta
     5461 gactctggct gcctgtatcg agtggtgatt ttgtgccgag ctgccggtcg gggagctgtt
     5521 gg
//
