LOCUS       sequence-393625-        7455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proUBQ10::PIP2A-mCherry-NOSt_pro35S::GFP-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7455)
  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 7455)
  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: JAB3421
FEATURES             Location/Qualifiers
     source          1..7455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..23
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     CDS             1546..2256
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQNKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMEELYK"
     primer_bind     complement(1691..1709)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      2275..2527
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        3050..3395
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     3338..3358
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     complement(3447..3475)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4070..4090
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4461..4481)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4461..4479)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4500..4516)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4500..4516)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4513..4535)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4524..4540
                     /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(4548..4554,4555..4572,4573..4578))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4593..4614
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4731..4748)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4853..4877
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4968..5556)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5048..5067)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5647..6435)
                     /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      6893..7328
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 tggcaggata tattgtggtg taacgttaag ctttggttgt cgacgagtca gtaataaacg
       61 gcgtcaaagt ggttgcagcc ggcacacacg agtcgtgttt atcaactcaa agcacaaata
      121 cttttcctca acctaaaaat aaggcaatta gccaaaaaca actttgcgtg taaacaacgc
      181 tcaatacacg tgtcatttta ttattagcta ttgcttcacc gccttagctt tctcgtgacc
      241 tagtcgtcct cgtcttttct tcttcttctt ctataaaaca atacccaaag agctcttctt
      301 cttcacaatt cagatttcaa tttctcaaaa tcttaaaaac tttctctcaa ttctctctac
      361 cgtgatcaag gtaaatttct gtgttcctta ttctctcaaa atcttcgatt ttgttttcgt
      421 tcgatcccaa tttcgtatat gttctttggt ttagattctg ttaatcttag atcgtagacg
      481 attttctggg tttgatcgtt agatatcatc ttaattctcg attagggttt catagatatc
      541 atccgatttg ttcaaataat ttgagttttg tcgaataatt actcttcgat ttgtgatttc
      601 tatctagatc tggtgttagt ttctagtttg tgcgatcgaa tttgtcgatt aatctgagtt
      661 tttctgatta acagcaccat ggcaaaggat gtggaagccg ttcccggaga aggatttcag
      721 acaagagact atcaagatcc gccaccagct ccgtttattg atggagcgga gctaaagaag
      781 tggtctttct acagagcagt tatcgcagag ttcgtagcca ctctcctctt cttatacatc
      841 accgttttga cagtcatcgg ttacaagatt cagtccgata ctgatgccgg tggcgtagat
      901 tgcggcggag ttggaatcct cggtatcgct tgggcctttg gtggtatgat cttcatcctc
      961 gtctactgca ccgccggtat ctctggtggt cacattaacc cagcggtgac atttgggcta
     1021 ttcttggcac gtaaagtgtc gttacctagg gccctattgt acataatcgc tcagtgtttg
     1081 ggtgcgattt gtggagttgg ttttgtcaaa gccttccaaa gctcttacta cacccgttac
     1141 ggaggtggag ccaactctct agccgatggc tacagcacag ggaccggtct agccgcagag
     1201 atcattggta ctttcgttct tgtctacacc gtattctctg ccactgaccc caaacgtagt
     1261 gccagagact cccacgttcc ggtgttggcg ccacttccaa tcggatttgc cgtgttcatg
     1321 gtacatttgg ctaccattcc cattaccgga accggaatta acccggcaag gagtttcgga
     1381 gctgccgtaa tctacaacaa gagcaagcca tgggatgacc actggatatt ttgggttgga
     1441 ccattcattg gagctgcgat agctgcattc taccaccaat tcgttctaag agcttcaggt
     1501 tctaagtctc ttggatcatt cagaagtgct gccaacgtcg gatccatggt gagcaagggc
     1561 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
     1621 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     1681 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
     1741 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
     1801 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     1861 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
     1921 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaataagacc
     1981 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
     2041 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
     2101 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac
     2161 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
     2221 cactccaccg gcggcatgga ggagctgtac aagtaaagcg gccgcccggc tgcagatcgt
     2281 tcaaacattt ggcaataaag tttcttaaga ttgaatcctg ttgccggtct tgcgatgatt
     2341 atcatataat ttctgttgaa ttacgttaag catgtaataa ttaacatgta atgcatgacg
     2401 ttatttatga gatgggtttt tatgattaga gtcccgcaat tatacattta atacgcgata
     2461 gaaaacaaaa tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc atctatgtta
     2521 ctagatcggg gcgcgccctc gaggaagacg taatgggtcc ccagatttgc cttttcaatt
     2581 tcagaaagaa tgctaaccca cagatggtta gagaggctta cgcagcagct ctcatcaaga
     2641 cgatctaccc gagcaataat ctccaggaaa tcaaatacct tcccaagaag gttaaagatg
     2701 cagtcaaaag attcaggact aactgcatca agaacacaga gaaagatata tttctcaaga
     2761 tcagaagtac tattccagta tggacgattc aaggcttgct tcacaaacca aggcaagtaa
     2821 tagagattgg agtctctaaa aaggtagttc ccactgaatc aaaggccatg gagtcaaaga
     2881 ttcaaataga ggacctaaca gaactcgccg taaagactgg cgaacagttc atacagagtc
     2941 tcttacgact caatgacaag aagaaaatct tcgtcaacat ggtggagcac gacacacttg
     3001 tctactccaa aaatatcaaa gatacagtct cagaagacca aagggcaatt gagacttttc
     3061 aacaaagggt aatatccgga aacctcctcg gattccattg cccagctatc tgtcacttta
     3121 ttgtgaagat agtggaaaag gaaggtggct cctacaaatg ccatcattgc gataaaggaa
     3181 aggccatcgt tgaagatgcc tctgccgaca gtggtcccaa agatggaccc ccacccacga
     3241 ggagcatcgt ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg gattgatgtg
     3301 atatctccac tgacgtaagg gatgacgcac aatcccacta tccttcgcaa gacccttcct
     3361 ctatataagg aagttcattt catttggaga gaacacgggg gactctagac accatggtga
     3421 gtaaaggaga agaacttttc actggagttg tcccaattct tgttgaatta gatggtgatg
     3481 ttaatgggca caaattttct gtcagtggag agggtgaagg tgatgcaaca tacggaaaac
     3541 ttacccttaa atttatttgc actactggaa aactacctgt tccatggcca accctggtca
     3601 ccaccctgac ctacggcgtg cagtgcttct cccgttaccc tgatcatatg aagcggcacg
     3661 acttcttcaa gagcgccatg cctgagggat acgtgcagga gaggaccatc ttcttcaagg
     3721 acgacgggaa ctacaagaca cgtgctgaag tcaagtttga gggagacacc ctcgtcaaca
     3781 ggatcgagct taagggaatc gatttcaagg aggacggaaa catcctcggc cacaagttgg
     3841 aatacaacta caactcccac aacgtataca tcatggccga caagcaaaag aacggcatca
     3901 aagccaactt caagacccgc cacaacatcg aagacggcgg cgtgcaactc gctgatcatt
     3961 atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt accagacaac cattacctgt
     4021 ccacacaatc tgccctttcg aaagatccca acgaaaagag agaccacatg gtccttcttg
     4081 agtttgtaac agctgctggg attacacatg gcatggatga actatacaaa taatgtcaag
     4141 ccattctctc gcagatgatg ttcactttgt gttttacttc ctttatgcat tcacagcaat
     4201 aaaagaaaga aatctccatc gcttttggtt ttcttctctg tcttaagtta gtcgttttcg
     4261 tgtctaatct attacttatc attgtaatag actcttcttc tattgagatt tgaatataaa
     4321 ctaaaacaca ttccatttta ctgtgttctc aacattcaga atgcaaacgg actaaccgta
     4381 gtgactcggc ttgagtcttc gggctagagc ggactagttc tagagcggcc gccaccgcgg
     4441 tggagctcca gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg
     4501 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4561 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4621 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4681 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4741 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4801 atacggttat ccacagaatc aggggataac gcaggaaaga acatgaaggc cttgacagga
     4861 tatattggcg ggtaaactaa rwcgctgtat gtgtttgttt gagatctcat gtgagcaaaa
     4921 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4981 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5041 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5101 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5161 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5221 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5281 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5341 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5401 actagaagaa cagtatttgg tatctgtgct ctgctgaagc cagttacctt cggaagaaga
     5461 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5521 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5581 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gaattcttgt
     5641 tagacattat ttgccgacta ccttggtgat ctcgcctttc acgtagtgaa caaattcttc
     5701 caactgatct gcgcgcgagg ccaagcgatc ttcttgtcca agataagcct gcctagcttc
     5761 aagtatgacg ggctgatact gggccggcag gcgctccatt gcccagtcgg cagcgacatc
     5821 cttcggcgcg attttgccgg ttactgcgct gtaccaaatg cgggacaacg taagcactac
     5881 atttcgctca tcgccagccc agtcgggcgg cgagttccat agcgttaagg tttcatttag
     5941 cgcctcaaat agatcctgtt caggaaccgg atcaaagagt tcctccgccg ctggacctac
     6001 caaggcaacg ctatgttctc ttgcttttgt cagcaagata gccagatcaa tgtcgatcgt
     6061 ggctggctcg aagatacctg caagaatgtc attgcgctgc cattctccaa attgcagttc
     6121 gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc acaacaatgg tgacttctac
     6181 agcgcggaga atctcgctct ctccagggga agccgaagtt tccaaaaggt cgttgatcaa
     6241 agctcgccgc gttgtttcat caagccttac ggtcaccgta accagcaaat caatatcact
     6301 gtgtggcttc aggccgccat ccactgcgga gccgtacaaa tgtacggcca gcaacgtcgg
     6361 ttcgagatgg cgctcgatga cgccaactac ctctgatagt tgagtcgata cttcggcgat
     6421 caccgcttcc ctcatgatgt ttaactcctg aattaagccg cgccgcgaag cggtgtcggc
     6481 ttgaatgaat tgttaggcgt catcctgtgc tcccgagaac cagtaccagt acatcgctgt
     6541 ttcgttcgag acttgaggtc tagttttata cgtgaacagg tcaatgccgc cgagagtaaa
     6601 gccacatttt gcgtacaaat tgcaggcagg tacattgttc gtttgtgcca tgatgatata
     6661 tttttatctt gtgcaatgta acatcagaga ttttgagaca caacgtggct ttgttgaata
     6721 aatcgaactt ttgctgagtt gaaggatcag atcacgcatc ttcccgacaa cgcagaccgt
     6781 tccgtggcaa agcaaaagtt caaaatcacc aactggtcca cctacaacaa agctctcatc
     6841 aaccgtggct ccctcacttt ctggctggat gatggggcga ttcaggagat ccccatccaa
     6901 cagcccgccg tcgagcgggc ttttttatcc ccggaagcct gtggatagag ggtagttatc
     6961 cacgtgaaac cgctaatgcc ccgcaaagcc ttgattcacg gggctttccg gcccgctcca
     7021 aaaactatcc acgtgaaatc gctaatcagg gtacgtgaaa tcgctaatcg gagtacgtga
     7081 aatcgctaat aaggtcacgt gaaatcgcta atcaaaaagg cacgtgagaa cgctaatagc
     7141 cctttcagat caacagcttg caaacacccc tcgctccggc aagtagttac agcaagtagt
     7201 atgttcaatt agcttttcaa ttatgaatat atatatcaat tattggtcgc ccttggcttg
     7261 tggacaatgc gctacgcgca ccggctccgc ccgtggacaa ccgcaagcgg ttgcccaccg
     7321 tcgagcgcct ttgcccacaa cccggcggcc ggccgcaaca gatcgtttta taaatttttt
     7381 tttttgaaaa agaaaaagcc cgaaaggcgg caacctctcg ggcttctgga tttccgatcc
     7441 ccggaattag actct
//
