LOCUS       sequence-389391-        7686 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants 
            (proUBQ10::PIP2A-mCherry-NOSt_pro35S-AmtRO-2XPhlFO::GFPint-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7686)
  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 7686)
  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: JAB2098
FEATURES             Location/Qualifiers
     source          1..7686
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     184..212
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     terminator      complement(1174..1426)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(1445..2155)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMEELYK"
     primer_bind     complement(1723..1742)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1992..2010
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     misc_feature    3678..3700
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     rep_origin      3828..4263
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     CDS             4721..5509
                     /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      5600..6188
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6089..6108
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6279..6303
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     6408..6425
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6542..6563
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6578..6583,6584..6601,6602..6608)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6616..6632
                     /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     6621..6643
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6640..6656
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6640..6656
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6675..6695
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6677..6695
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7066..7086)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     intron          join(7542..7686,1..44)
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
ORIGIN
        1 tgataattat tatatatata tcaaaggtag aagcagaaac ttacgtaggt cagggtggtg
       61 accagggttg gccatggaac aggtagtttt ccagtagtgc aaataaattt aagggtaagt
      121 tttccgtatg ttgcatcacc ttcaccctct ccactgacag aaaatttgtg cccattaaca
      181 tcaccatcta attcaacaag aattgggaca actccagtga aaagttcttc tcctttactc
      241 accatggtgt ctagagtccc ccgtgtacct taacgatacg gtacgtttcg tatcaatcca
      301 ccttaacgat acggtacgtt tcgtatcaat tatatagatt atctatagat cgatagaaga
      361 tagtgggatt gtgcgtcatc ccttacgtca gtggagatat cacatcaatc cacttgcttt
      421 gaagacgtgg ttggaacgtc ttctttttcc acgatgctcc tcgtgggtgg gggtccatct
      481 ttgggaccac tgtcggcaga ggcatcttca acgatggcct ttcctttatc gcaatgatgg
      541 catttgtagg agccaccttc cttttccact atcttcacaa taaagtgaca gatagctggg
      601 caatggaatc cgaggaggtt tccggatatt accctttgtt gaaaagtctc aattgccctt
      661 tggtcttctg agactgtatc tttgatattt ttggagtaga caagtgtgtc gtgctccacc
      721 atgttgacga agattttctt cttgtcattg agtcgtaaga gactctgtat gaactgttcg
      781 ccagtcttta cggcgagttc tgttaggtcc tctatttgaa tctttgactc catggccttt
      841 gattcagtgg gaactacctt tttagagact ccaatctcta ttacttgcct tggtttgtga
      901 agcaagcctt gaatcgtcca tactggaata gtacttctga tcttgagaaa tatatctttc
      961 tctgtgttct tgatgcagtt agtcctgaat cttttgactg catctttaac cttcttggga
     1021 aggtatttga tttcctggag attattgctc gggtagatcg tcttgatgag agctgctgcg
     1081 taagcctctc tacccatctg tgggttagca ttctttctga aattgaaaag gcaaatctgg
     1141 ggacccatta cgtctccctc gagggcgcgc cccgatctag taacatagat gacaccgcgc
     1201 gcgataattt atcctagttt gcgcgctata ttttgttttc tatcgcgtat taaatgtata
     1261 attgcgggac tctaatcata aaaacccatc tcataaataa cgtcatgcat tacatgttaa
     1321 ttattacatg cttaacgtaa ttcaacagaa attatatgat aatcatcgca agaccggcaa
     1381 caggattcaa tcttaagaaa ctttattgcc aaatgtttga acgatctgca gccgggcggc
     1441 cgctttactt gtacagctcc tccatgccgc cggtggagtg gcggccctcg gcgcgttcgt
     1501 actgttccac gatggtgtag tcctcgttgt gggaggtgat gtccaacttg atgttgacgt
     1561 tgtaggcgcc gggcagctgc acgggcttct tggccttgta ggtggtcttg acctcagcgt
     1621 cgtagtggcc gccgtccttc agcttcagcc tctgcttgat ctcgcccttc agggcgccgt
     1681 cctcggggta catccgctcg gaggaggcct cccagcccat ggtcttcttc tgcattacgg
     1741 ggccgtcgga ggggaagttg gtgccgcgca gcttcacctt gtagatgaac tcgccgtcct
     1801 gcagggagga gtcctgggtc acggtcacca cgccgccgtc ctcgaagttc atcacgcgct
     1861 cccacttgaa gccctcgggg aaggacagct tcaagtagtc ggggatgtcg gcggggtgct
     1921 tcacgtaggc cttggagccg tacatgaact gaggggacag gatgtcccag gcgaagggca
     1981 gggggccacc cttggtcacc ttcagcttgg cggtctgggt gccctcgtag gggcggccct
     2041 cgccctcgcc ctcgatctcg aactcgtggc cgttcacgga gccctccatg tgcaccttga
     2101 agcgcatgaa ctccttgatg atggccatgt tatcctcctc gcccttgctc accatggatc
     2161 cgacgttggc agcacttctg aatgatccaa gagacttaga acctgaagct cttagaacga
     2221 attggtggta gaatgcagct atcgcagctc caatgaatgg tccaacccaa aatatccagt
     2281 ggtcatccca tggcttgctc ttgttgtaga ttacggcagc tccgaaactc cttgccgggt
     2341 taattccggt tccggtaatg ggaatggtag ccaaatgtac catgaacacg gcaaatccga
     2401 ttggaagtgg cgccaacacc ggaacgtggg agtctctggc actacgtttg gggtcagtgg
     2461 cagagaagac ggtgtagaca agaacgaaag taccaatgat ctctgcggct agaccggtcc
     2521 ctgtgctgta gccatcggct agagagttgg ctccacctcc gtaacgggtg tagtaagagc
     2581 tttggaaggc tttgacaaaa ccaactccac aaatcgcacc caaacactga gcgattatgt
     2641 acaatagggc cctaggtaac gacactttac gtgccaagaa tagcccaaat gtcaccgctg
     2701 ggttaatgtg accaccagag ataccggcgg tgcagtagac gaggatgaag atcataccac
     2761 caaaggccca agcgataccg aggattccaa ctccgccgca atctacgcca ccggcatcag
     2821 tatcggactg aatcttgtaa ccgatgactg tcaaaacggt gatgtataag aagaggagag
     2881 tggctacgaa ctctgcgata actgctctgt agaaagacca cttctttagc tccgctccat
     2941 caataaacgg agctggtggc ggatcttgat agtctcttgt ctgaaatcct tctccgggaa
     3001 cggcttccac atcctttgcc atggtgctgt taatcagaaa aactcagatt aatcgacaaa
     3061 ttcgatcgca caaactagaa actaacacca gatctagata gaaatcacaa atcgaagagt
     3121 aattattcga caaaactcaa attatttgaa caaatcggat gatatctatg aaaccctaat
     3181 cgagaattaa gatgatatct aacgatcaaa cccagaaaat cgtcttcgat ctaagattaa
     3241 cagaatctaa accaaagaac atatacgaaa ttgggatcga acgaaaacaa aatcgaagat
     3301 tttgagagaa taaggaacac agaaatttac cttgatcacg gtagagagaa ttgagagaaa
     3361 gtttttaaga ttttgagaaa ttgaaatctg aattgtgaag aagaagagct ctttgggtat
     3421 tgttttatag aagaagaaga agaaaagacg aggacgacta ggtcacgaga aagctaaggc
     3481 ggtgaagcaa tagctaataa taaaatgaca cgtgtattga gcgttgttta cacgcaaagt
     3541 tgtttttggc taattgcctt atttttaggt tgaggaaaag tatttgtgct ttgagttgat
     3601 aaacacgact cgtgtgtgcc ggctgcaacc actttgacgc cgtttattac tgactcgtcg
     3661 acaaccaaag cttaacgtta caccacaata tatcctgcca agagtctaat tccggggatc
     3721 ggaaatccag aagcccgaga ggttgccgcc tttcgggctt tttctttttc aaaaaaaaaa
     3781 atttataaaa cgatctgttg cggccggccg ccgggttgtg ggcaaaggcg ctcgacggtg
     3841 ggcaaccgct tgcggttgtc cacgggcgga gccggtgcgc gtagcgcatt gtccacaagc
     3901 caagggcgac caataattga tatatatatt cataattgaa aagctaattg aacatactac
     3961 ttgctgtaac tacttgccgg agcgaggggt gtttgcaagc tgttgatctg aaagggctat
     4021 tagcgttctc acgtgccttt ttgattagcg atttcacgtg accttattag cgatttcacg
     4081 tactccgatt agcgatttca cgtaccctga ttagcgattt cacgtggata gtttttggag
     4141 cgggccggaa agccccgtga atcaaggctt tgcggggcat tagcggtttc acgtggataa
     4201 ctaccctcta tccacaggct tccggggata aaaaagcccg ctcgacggcg ggctgttgga
     4261 tggggatctc ctgaatcgcc ccatcatcca gccagaaagt gagggagcca cggttgatga
     4321 gagctttgtt gtaggtggac cagttggtga ttttgaactt ttgctttgcc acggaacggt
     4381 ctgcgttgtc gggaagatgc gtgatctgat ccttcaactc agcaaaagtt cgatttattc
     4441 aacaaagcca cgttgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat
     4501 catcatggca caaacgaaca atgtacctgc ctgcaatttg tacgcaaaat gtggctttac
     4561 tctcggcggc attgacctgt tcacgtataa aactagacct caagtctcga acgaaacagc
     4621 gatgtactgg tactggttct cgggagcaca ggatgacgcc taacaattca ttcaagccga
     4681 caccgcttcg cggcgcggct taattcagga gttaaacatc atgagggaag cggtgatcgc
     4741 cgaagtatcg actcaactat cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac
     4801 gttgctggcc gtacatttgt acggctccgc agtggatggc ggcctgaagc cacacagtga
     4861 tattgatttg ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc gagctttgat
     4921 caacgacctt ttggaaactt cggcttcccc tggagagagc gagattctcc gcgctgtaga
     4981 agtcaccatt gttgtgcacg acgacatcat tccgtggcgt tatccagcta agcgcgaact
     5041 gcaatttgga gaatggcagc gcaatgacat tcttgcaggt atcttcgagc cagccacgat
     5101 cgacattgat ctggctatct tgctgacaaa agcaagagaa catagcgttg ccttggtagg
     5161 tccagcggcg gaggaactct ttgatccggt tcctgaacag gatctatttg aggcgctaaa
     5221 tgaaacctta acgctatgga actcgccgcc cgactgggct ggcgatgagc gaaatgtagt
     5281 gcttacgttg tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt
     5341 cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca tacttgaagc
     5401 taggcaggct tatcttggac aagaagatcg cttggcctcg cgcgcagatc agttggaaga
     5461 atttgttcac tacgtgaaag gcgagatcac caaggtagtc ggcaaataat gtctaacaag
     5521 aattcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5581 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5641 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttc
     5701 ttccgaaggt aactggcttc agcagagcac agataccaaa tactgttctt ctagtgtagc
     5761 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5821 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5881 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5941 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6001 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     6061 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6121 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6181 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6241 ctcacatgag atctcaaaca aacacataca gcgacttagt ttacccgcca atatatcctg
     6301 tcaaggcctt catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     6361 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     6421 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     6481 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     6541 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     6601 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     6661 attacgccaa gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctccaccgcg
     6721 gtggcggccg ctctagaact agtccgctct agcccggaga ctcaagccga gtcactacgg
     6781 ttagtccgtt tgcattctga atgttgagaa cacagtaaaa tggaatgtgt tttagtttat
     6841 attcaaatct caatagaaga agagtctatt acaatgataa gtaatagatt agacacgaaa
     6901 acgactaact taagacagag aagaaaacca aaagcgatgg agatttcttt cttttattgc
     6961 tgtgaatgca taaaggaagt aaaacacaaa gtgaacatca tctgcgagag aatggcttga
     7021 cattatttgt atagttcatc catgccatgt gtaatcccag cagctgttac aaactcaaga
     7081 aggaccatgt gatctctctt ttcgttggga tctttcgaaa gggcagattg tgtggacagg
     7141 taatggttgt ctggtaaaag gacagggcca tcgccaattg gagtattttg ttgataatga
     7201 tcagcgagtt gcacgccgcc gtcttcgatg ttgtggcggg tcttgaagtt ggctttgatg
     7261 ccgttctttt gcttgtcggc catgatgtat acgttgtggg agttgtagtt gtattccaac
     7321 ttgtggccga ggatgtttcc gtcctccttg aaatcgattc ccttaagctc gatcctgttg
     7381 acgagggtgt caccctcaaa cttgacttca gcacgtgtct tgtagttccc gtcgtccttg
     7441 aagaagatgg tcctctcctg cacgtatccc tcaggcatgg cgctcttgaa gaagtcgtgc
     7501 cgcttcatat gatcagggta acgggagaag cactgcacgc cctgcacatc aacaaatttt
     7561 ggtcatatat tagaaaagtt ataaattaaa atatacacac ttataaacta cagaaaagca
     7621 attgctatat actacattct tttattttga aaaaaatatt tgaaatatta tattactact
     7681 aattaa
//
