LOCUS       sequence-387106-        5647 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (6XAmtRO-2XPhlFO::TetR-NLS-19St).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5647)
  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 5647)
  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: JAB2964
FEATURES             Location/Qualifiers
     source          1..5647
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    27..51
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(121..373)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(393..944)
                     /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(986..1165)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        1559..1605
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             2281..2301
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2357..2533
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(2615..2635)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2615..2633)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2654..2670)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2654..2670)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2667..2689)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2678..2694
                     /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(2702..2708,2709..2726,2727..2732))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2747..2768
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2885..2902)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3007..3031
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(3122..3710)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3202..3221)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3801..4589)
                     /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      5047..5482
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 cggtcgggga gctgttggct ggctggtggc aggatatatt gtggtgtaaa caaattgacg
       61 cttagacaac ttaataacac attgcggacg tttttaatgt aatcagcttg catgccggtc
      121 gatctagtaa catagatgac accgcgcgcg ataatttatc ctagtttgcg cgctatattt
      181 tgttttctat cgcgtattaa atgtataatt gcgggactct aatcataaaa acccatctca
      241 taaataacgt catgcattac atgttaatta ttacatgctt aacgtaattc aacagaaatt
      301 atatgataat catcgcaaga ccggcaacag gattcaatct taagaaactt tattgccaaa
      361 tgtttgaacg atctgcttga ctctaggggt catcagattt cggtgacggg caggaccgga
      421 cggggcggca ccggcaggct gaagtccagc tgccagaaac ccacgtcatg ccagttcccg
      481 tgcttgaagc cggccgcccg cagcatgccg cggggggcat atccgagcgc ctcgtgcatg
      541 cgcacgctcg ggtcgttggg cagcccgatg acagcgacca cgctcttgaa gccctgtgcc
      601 tccagggact tcagcaggtg ggtgtagagc gtggagccca gtcccgtccg ctggtggcgg
      661 ggggacacgt acacggtcga ctcggccgtc cagtcgtagg cgttgcgtgc cttccaggga
      721 cccgcgtagg cgatgccggc gacctcgccg tccacctcgg cgacgagcca gggatagcgc
      781 tcccgcagac ggacgaggtc gtccgtccac tcctgcggtt cctgcggctc ggtacggaag
      841 ttgaccgtgc ttgtctcgat gtagtggttg acgatggtgc agaccgccgg catgtccgcc
      901 tcggtggcac ggcggatgtc ggccgggcgt cgttctgggc tcatggtaga tcccctcgat
      961 cgagttgaga gtgaatatga gactctaatt ggataccgag gggaatttat ggaacgtcag
     1021 tggagcattt ttgacaagaa atatttgcta gctgatagtg accttaggcg acttttgaac
     1081 gcgcaataat ggtttctgac gtatgtgctt agctcattaa actccagaaa cccgcggctc
     1141 agtggctcct tcaacgttgc ggttctgtca gttccaaacg taaaacggct tgtcccgcgt
     1201 catcggcggg ggtcataacg tgactccctt aattctccgc tcatgatcga taacatgtcg
     1261 actatcggac gagctcaaag tttaacgtgt gaagacgtgc ttctaatgat agaaaagttg
     1321 gctgcggccg cgaatttcta tcgatctata gataatgatt tctatcgatc tatagataat
     1381 catttctatc gatctataga taatcatttc tatcgatcta tagataatgt tttctatcga
     1441 tctatagata atagtttcta tcgatctata gataatcggg tctaagctag ctaggccacc
     1501 tagcatgata cgaaacgtac cgtatcgtta aggtaggtca caatcccact atcctttcgc
     1561 aagacccttc ctctatataa ggaagttcat ttcatttgga gagaacacgg gggactctag
     1621 agtaattgat acgaaacgta ccgtatcgtt aaggtcacca tgagccggct ggacaagagc
     1681 aaagtgatca acagcgccct ggaactgctc aacgaagtgg gcatcgaggg cctgaccacc
     1741 cggaagctgg ctcagaaact gggcgtggaa cagcccaccc tgtactggca cgtgaagaac
     1801 aagagagccc tgctggacgc cctggccatc gagatgctgg accggcacca cacccacttt
     1861 tgccccctgg aaggcgagag ctggcaggat ttcctgcgga acaacgccaa gagcttcaga
     1921 tgcgctctgc tgtcccaccg ggatggcgcc aaagtgcacc tgggcaccag acctaccgag
     1981 aagcagtacg agacactgga aaaccagctg gccttcctgt gccagcaggg cttcagcctg
     2041 gaaaatgccc tgtacgccct gagcgccgtg ggccacttta cactgggctg cgtgctggaa
     2101 gatcaggaac accaggtcgc caaagaggaa agagagacac ccaccaccga cagcatgccc
     2161 cctctgctga gacaggccat tgagctgttc gatcatcagg gcgccgagcc cgcctttctg
     2221 ttcggcctcg agctgatcat ctgtggcctc gaaaaacagc tgaagtgcga gtccggcagc
     2281 ccgaagaaga agaggaaggt ttagtgtcgg tacgctgaaa tcaccagtct ctctctacaa
     2341 atctatctct ctctattttc tccataaata atgtgtgagt agtttcccga taagggaaat
     2401 tagggttctt atagggtttc gctcatgtgt tgagcatata agaaaccctt agtatgtatt
     2461 tgtatttgta aaatacttct atcaataaaa tttctaattc ctaaaaccaa aatccagtac
     2521 taaaatccag atcgatgctt gatactacgt cttcgggcta gagcggacta gttctagagc
     2581 ggccgccacc gcggtggagc tccagctttt gttcccttta gtgagggtta attgcgcgct
     2641 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     2701 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     2761 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     2821 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     2881 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     2941 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatga
     3001 aggccttgac aggatatatt ggcgggtaaa ctaagtcgct gtatgtgttt gtttgagatc
     3061 tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3121 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3181 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3241 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3301 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3361 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3421 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3481 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3541 ctaactacgg ctacactaga agaacagtat ttggtatctg tgctctgctg aagccagtta
     3601 ccttcggaag aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3661 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3721 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3781 tcatgaattc ttgttagaca ttatttgccg actaccttgg tgatctcgcc tttcacgtag
     3841 tgaacaaatt cttccaactg atctgcgcgc gaggccaagc gatcttcttg tccaagataa
     3901 gcctgcctag cttcaagtat gacgggctga tactgggccg gcaggcgctc cattgcccag
     3961 tcggcagcga catccttcgg cgcgattttg ccggttactg cgctgtacca aatgcgggac
     4021 aacgtaagca ctacatttcg ctcatcgcca gcccagtcgg gcggcgagtt ccatagcgtt
     4081 aaggtttcat ttagcgcctc aaatagatcc tgttcaggaa ccggatcaaa gagttcctcc
     4141 gccgctggac ctaccaaggc aacgctatgt tctcttgctt ttgtcagcaa gatagccaga
     4201 tcaatgtcga tcgtggctgg ctcgaagata cctgcaagaa tgtcattgcg ctgccattct
     4261 ccaaattgca gttcgcgctt agctggataa cgccacggaa tgatgtcgtc gtgcacaaca
     4321 atggtgactt ctacagcgcg gagaatctcg ctctctccag gggaagccga agtttccaaa
     4381 aggtcgttga tcaaagctcg ccgcgttgtt tcatcaagcc ttacggtcac cgtaaccagc
     4441 aaatcaatat cactgtgtgg cttcaggccg ccatccactg cggagccgta caaatgtacg
     4501 gccagcaacg tcggttcgag atggcgctcg atgacgccaa ctacctctga tagttgagtc
     4561 gatacttcgg cgatcaccgc ttccctcatg atgtttaact cctgaattaa gccgcgccgc
     4621 gaagcggtgt cggcttgaat gaattgttag gcgtcatcct gtgctcccga gaaccagtac
     4681 cagtacatcg ctgtttcgtt cgagacttga ggtctagttt tatacgtgaa caggtcaatg
     4741 ccgccgagag taaagccaca ttttgcgtac aaattgcagg caggtacatt gttcgtttgt
     4801 gccatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga gacacaacgt
     4861 ggctttgttg aataaatcga acttttgctg agttgaagga tcagatcacg catcttcccg
     4921 acaacgcaga ccgttccgtg gcaaagcaaa agttcaaaat caccaactgg tccacctaca
     4981 acaaagctct catcaaccgt ggctccctca ctttctggct ggatgatggg gcgattcagg
     5041 agatccccat ccaacagccc gccgtcgagc gggctttttt atccccggaa gcctgtggat
     5101 agagggtagt tatccacgtg aaaccgctaa tgccccgcaa agccttgatt cacggggctt
     5161 tccggcccgc tccaaaaact atccacgtga aatcgctaat cagggtacgt gaaatcgcta
     5221 atcggagtac gtgaaatcgc taataaggtc acgtgaaatc gctaatcaaa aaggcacgtg
     5281 agaacgctaa tagccctttc agatcaacag cttgcaaaca cccctcgctc cggcaagtag
     5341 ttacagcaag tagtatgttc aattagcttt tcaattatga atatatatat caattattgg
     5401 tcgcccttgg cttgtggaca atgcgctacg cgcaccggct ccgcccgtgg acaaccgcaa
     5461 gcggttgccc accgtcgagc gcctttgccc acaacccggc ggccggccgc aacagatcgt
     5521 tttataaatt tttttttttg aaaaagaaaa agcccgaaag gcggcaacct ctcgggcttc
     5581 tggatttccg atccccggaa ttagactctg gctgcctgta tcgagtggtg attttgtgcc
     5641 gagctgc
//
