LOCUS       sequence-387116-        6155 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (pro35S::TetR-NLS-19St).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6155)
  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 6155)
  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: JAB3617
FEATURES             Location/Qualifiers
     source          1..6155
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      182..617
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     CDS             1075..1863
                     /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      1954..2542
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2443..2462
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2633..2657
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     2762..2779
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2896..2917
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2932..2937,2938..2955,2956..2962)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2970..2986
                     /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)."
     primer_bind     2975..2997
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2994..3010
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2994..3010
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3029..3049
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3031..3049
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    3131..3307
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(3363..3383)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        5007..5186
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             5228..5779
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     terminator      5799..6051
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    6121..6145
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 aacagctccc cgaccggcag ctcggcacaa aatcaccact cgatacaggc agccagagtc
       61 taattccggg gatcggaaat ccagaagccc gagaggttgc cgcctttcgg gctttttctt
      121 tttcaaaaaa aaaaatttat aaaacgatct gttgcggccg gccgccgggt tgtgggcaaa
      181 ggcgctcgac ggtgggcaac cgcttgcggt tgtccacggg cggagccggt gcgcgtagcg
      241 cattgtccac aagccaaggg cgaccaataa ttgatatata tattcataat tgaaaagcta
      301 attgaacata ctacttgctg taactacttg ccggagcgag gggtgtttgc aagctgttga
      361 tctgaaaggg ctattagcgt tctcacgtgc ctttttgatt agcgatttca cgtgacctta
      421 ttagcgattt cacgtactcc gattagcgat ttcacgtacc ctgattagcg atttcacgtg
      481 gatagttttt ggagcgggcc ggaaagcccc gtgaatcaag gctttgcggg gcattagcgg
      541 tttcacgtgg ataactaccc tctatccaca ggcttccggg gataaaaaag cccgctcgac
      601 ggcgggctgt tggatgggga tctcctgaat cgccccatca tccagccaga aagtgaggga
      661 gccacggttg atgagagctt tgttgtaggt ggaccagttg gtgattttga acttttgctt
      721 tgccacggaa cggtctgcgt tgtcgggaag atgcgtgatc tgatccttca actcagcaaa
      781 agttcgattt attcaacaaa gccacgttgt gtctcaaaat ctctgatgtt acattgcaca
      841 agataaaaat atatcatcat ggcacaaacg aacaatgtac ctgcctgcaa tttgtacgca
      901 aaatgtggct ttactctcgg cggcattgac ctgttcacgt ataaaactag acctcaagtc
      961 tcgaacgaaa cagcgatgta ctggtactgg ttctcgggag cacaggatga cgcctaacaa
     1021 ttcattcaag ccgacaccgc ttcgcggcgc ggcttaattc aggagttaaa catcatgagg
     1081 gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc
     1141 catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg
     1201 aagccacaca gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg
     1261 cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt
     1321 ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca
     1381 gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc
     1441 gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc
     1501 gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta
     1561 tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat
     1621 gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc
     1681 gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc
     1741 gtcatacttg aagctaggca ggcttatctt ggacaagaag atcgcttggc ctcgcgcgca
     1801 gatcagttgg aagaatttgt tcactacgtg aaaggcgaga tcaccaaggt agtcggcaaa
     1861 taatgtctaa caagaattca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     1921 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     1981 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2041 gctaccaact cttcttccga aggtaactgg cttcagcaga gcacagatac caaatactgt
     2101 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2161 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2221 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2281 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2341 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2401 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     2461 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     2521 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     2581 ttgctggcct tttgctcaca tgagatctca aacaaacaca tacagcgact tagtttaccc
     2641 gccaatatat cctgtcaagg ccttcatgtt ctttcctgcg ttatcccctg attctgtgga
     2701 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     2761 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     2821 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     2881 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     2941 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     3001 cagctatgac catgattacg ccaagcgcgc aattaaccct cactaaaggg aacaaaagct
     3061 ggagctccac cgcggtggcg gccgctctag aactagtccg ctctagcccg aagacgtagt
     3121 atcaagcatc gatctggatt ttagtactgg attttggttt taggaattag aaattttatt
     3181 gatagaagta ttttacaaat acaaatacat actaagggtt tcttatatgc tcaacacatg
     3241 agcgaaaccc tataagaacc ctaatttccc ttatcgggaa actactcaca cattatttat
     3301 ggagaaaata gagagagata gatttgtaga gagagactgg tgatttcagc gtaccgacac
     3361 taaaccttcc tcttcttctt cgggctgccg gactcgcact tcagctgttt ttcgaggcca
     3421 cagatgatca gctcgaggcc gaacagaaag gcgggctcgg cgccctgatg atcgaacagc
     3481 tcaatggcct gtctcagcag agggggcatg ctgtcggtgg tgggtgtctc tctttcctct
     3541 ttggcgacct ggtgttcctg atcttccagc acgcagccca gtgtaaagtg gcccacggcg
     3601 ctcagggcgt acagggcatt ttccaggctg aagccctgct ggcacaggaa ggccagctgg
     3661 ttttccagtg tctcgtactg cttctcggta ggtctggtgc ccaggtgcac tttggcgcca
     3721 tcccggtggg acagcagagc gcatctgaag ctcttggcgt tgttccgcag gaaatcctgc
     3781 cagctctcgc cttccagggg gcaaaagtgg gtgtggtgcc ggtccagcat ctcgatggcc
     3841 agggcgtcca gcagggctct cttgttcttc acgtgccagt acagggtggg ctgttccacg
     3901 cccagtttct gagccagctt ccgggtggtc aggccctcga tgcccacttc gttgagcagt
     3961 tccagggcgc tgttgatcac tttgctcttg tccagccggc tcatggtgtc tagagtcccc
     4021 cgtgttctct ccaaatgaaa tgaacttcct tatatagaat tatctatctt gcgaaggata
     4081 gtgggattgt gcgtcatccc ttacgtcagt ggagatatca catcaatcca cttgctttgt
     4141 agacgtggtt ggaacgtcta ctttttccac gatgctcctc gtgggtgggg gtccatcttt
     4201 gggaccactg tcggcagagg catcttcaac gatggccttt cctttatcgc aatgatggca
     4261 tttgtaggag ccaccttcct tttccactat cttcacaata aagtgacaga tagctgggca
     4321 atggaatccg aggaggtttc cggatattac cctttgttga aaagtctcaa ttgccctttg
     4381 gtctactgag actgtatctt tgatattttt ggagtagaca agtgtgtcgt gctccaccat
     4441 gttgacgaag attttcttct tgtcattgag tcgtaagaga ctctgtatga actgttcgcc
     4501 agtctttacg gcgagttctg ttaggtcctc tatttgaatc tttgactcca tggcctttga
     4561 ttcagtggga actacctttt tagagactcc aatctctatt acttgccttg gtttgtgaag
     4621 caagccttga atcgtccata ctggaatagt acttctgatc ttgagaaata tatctttctc
     4681 tgtgttcttg atgcagttag tcctgaatct tttgactgca tctttaacct tcttgggaag
     4741 gtatttgatt tcctggagat tattgctcgg gtagatcgtc ttgatgagag ctgctgcgta
     4801 agcctctcta accatctgtg ggttagcatt ctttctgaaa ttgaaaaggc aaatctgggg
     4861 acccattaga agcacgtctt cacacgttaa actttgagct cgtccgatag tcgacatgtt
     4921 atcgatcatg agcggagaat taagggagtc acgttatgac ccccgccgat gacgcgggac
     4981 aagccgtttt acgtttggaa ctgacagaac cgcaacgttg aaggagccac tgagccgcgg
     5041 gtttctggag tttaatgagc taagcacata cgtcagaaac cattattgcg cgttcaaaag
     5101 tcgcctaagg tcactatcag ctagcaaata tttcttgtca aaaatgctcc actgacgttc
     5161 cataaattcc cctcggtatc caattagagt ctcatattca ctctcaactc gatcgagggg
     5221 atctaccatg agcccagaac gacgcccggc cgacatccgc cgtgccaccg aggcggacat
     5281 gccggcggtc tgcaccatcg tcaaccacta catcgagaca agcacggtca acttccgtac
     5341 cgagccgcag gaaccgcagg agtggacgga cgacctcgtc cgtctgcggg agcgctatcc
     5401 ctggctcgtc gccgaggtgg acggcgaggt cgccggcatc gcctacgcgg gtccctggaa
     5461 ggcacgcaac gcctacgact ggacggccga gtcgaccgtg tacgtgtccc cccgccacca
     5521 gcggacggga ctgggctcca cgctctacac ccacctgctg aagtccctgg aggcacaggg
     5581 cttcaagagc gtggtcgctg tcatcgggct gcccaacgac ccgagcgtgc gcatgcacga
     5641 ggcgctcgga tatgcccccc gcggcatgct gcgggcggcc ggcttcaagc acgggaactg
     5701 gcatgacgtg ggtttctggc agctggactt cagcctgccg gtgccgcccc gtccggtcct
     5761 gcccgtcacc gaaatctgat gacccctaga gtcaagcaga tcgttcaaac atttggcaat
     5821 aaagtttctt aagattgaat cctgttgccg gtcttgcgat gattatcata taatttctgt
     5881 tgaattacgt taagcatgta ataattaaca tgtaatgcat gacgttattt atgagatggg
     5941 tttttatgat tagagtcccg caattataca tttaatacgc gatagaaaac aaaatatagc
     6001 gcgcaaacta ggataaatta tcgcgcgcgg tgtcatctat gttactagat cgaccggcat
     6061 gcaagctgat tacattaaaa acgtccgcaa tgtgttatta agttgtctaa gcgtcaattt
     6121 gtttacacca caatatatcc tgccaccagc cagcc
//
