LOCUS       sequence-389442-        7704 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants 
            (proUBQ10::PIP2A-mCherry-NOSt_pro35S-PhlFO::GFPint-NLS-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7704)
  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 7704)
  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: JAB2296
FEATURES             Location/Qualifiers
     source          1..7704
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    22..44
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     CDS             1567..2277
                     /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(1712..1730)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      2296..2548
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        3071..3416
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     3359..3379
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     complement(3498..3526)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          3666..3854
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     4310..4330
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             4371..4391
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4731..4751)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4731..4749)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4770..4786)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4770..4786)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4783..4805)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4794..4810
                     /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(4818..4824,4825..4842,4843..4848))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4863..4884
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5001..5018)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5123..5147
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5238..5826)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5318..5337)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5917..6705)
                     /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      7163..7598
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 cgatccccgg aattagactc ttggcaggat atattgtggt gtaacgttaa gctttggttg
       61 tcgacgagtc agtaataaac ggcgtcaaag tggttgcagc cggcacacac gagtcgtgtt
      121 tatcaactca aagcacaaat acttttcctc aacctaaaaa taaggcaatt agccaaaaac
      181 aactttgcgt gtaaacaacg ctcaatacac gtgtcatttt attattagct attgcttcac
      241 cgccttagct ttctcgtgac ctagtcgtcc tcgtcttttc ttcttcttct tctataaaac
      301 aatacccaaa gagctcttct tcttcacaat tcagatttca atttctcaaa atcttaaaaa
      361 ctttctctca attctctcta ccgtgatcaa ggtaaatttc tgtgttcctt attctctcaa
      421 aatcttcgat tttgttttcg ttcgatccca atttcgtata tgttctttgg tttagattct
      481 gttaatctta gatcgtagac gattttctgg gtttgatcgt tagatatcat cttaattctc
      541 gattagggtt tcatagatat catccgattt gttcaaataa tttgagtttt gtcgaataat
      601 tactcttcga tttgtgattt ctatctagat ctggtgttag tttctagttt gtgcgatcga
      661 atttgtcgat taatctgagt ttttctgatt aacagcacca tggcaaagga tgtggaagcc
      721 gttcccggag aaggatttca gacaagagac tatcaagatc cgccaccagc tccgtttatt
      781 gatggagcgg agctaaagaa gtggtctttc tacagagcag ttatcgcaga gttcgtagcc
      841 actctcctct tcttatacat caccgttttg acagtcatcg gttacaagat tcagtccgat
      901 actgatgccg gtggcgtaga ttgcggcgga gttggaatcc tcggtatcgc ttgggccttt
      961 ggtggtatga tcttcatcct cgtctactgc accgccggta tctctggtgg tcacattaac
     1021 ccagcggtga catttgggct attcttggca cgtaaagtgt cgttacctag ggccctattg
     1081 tacataatcg ctcagtgttt gggtgcgatt tgtggagttg gttttgtcaa agccttccaa
     1141 agctcttact acacccgtta cggaggtgga gccaactctc tagccgatgg ctacagcaca
     1201 gggaccggtc tagccgcaga gatcattggt actttcgttc ttgtctacac cgtattctct
     1261 gccactgacc ccaaacgtag tgccagagac tcccacgttc cggtgttggc gccacttcca
     1321 atcggatttg ccgtgttcat ggtacatttg gctaccattc ccattaccgg aaccggaatt
     1381 aacccggcaa ggagtttcgg agctgccgta atctacaaca agagcaagcc atgggatgac
     1441 cactggatat tttgggttgg accattcatt ggagctgcga tagctgcatt ctaccaccaa
     1501 ttcgttctaa gagcttcagg ttctaagtct cttggatcat tcagaagtgc tgccaacgtc
     1561 ggatccatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
     1621 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     1681 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
     1741 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
     1801 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
     1861 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
     1921 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     1981 cccgtaatgc agaataagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
     2041 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
     2101 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     2161 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
     2221 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg aggagctgta caagtaaagc
     2281 ggccgcccgg ctgcagatcg ttcaaacatt tggcaataaa gtttcttaag attgaatcct
     2341 gttgccggtc ttgcgatgat tatcatataa tttctgttga attacgttaa gcatgtaata
     2401 attaacatgt aatgcatgac gttatttatg agatgggttt ttatgattag agtcccgcaa
     2461 ttatacattt aatacgcgat agaaaacaaa atatagcgcg caaactagga taaattatcg
     2521 cgcgcggtgt catctatgtt actagatcgg ggcgcgccct cgaggaagac gtaatgggtc
     2581 cccagatttg ccttttcaat ttcagaaaga atgctaaccc acagatggtt agagaggctt
     2641 acgcagcagc tctcatcaag acgatctacc cgagcaataa tctccaggaa atcaaatacc
     2701 ttcccaagaa ggttaaagat gcagtcaaaa gattcaggac taactgcatc aagaacacag
     2761 agaaagatat atttctcaag atcagaagta ctattccagt atggacgatt caaggcttgc
     2821 ttcacaaacc aaggcaagta atagagattg gagtctctaa aaaggtagtt cccactgaat
     2881 caaaggccat ggagtcaaag attcaaatag aggacctaac agaactcgcc gtaaagactg
     2941 gcgaacagtt catacagagt ctcttacgac tcaatgacaa gaagaaaatc ttcgtcaaca
     3001 tggtggagca cgacacactt gtctactcca aaaatatcaa agatacagtc tcagaagacc
     3061 aaagggcaat tgagactttt caacaaaggg taatatccgg aaacctcctc ggattccatt
     3121 gcccagctat ctgtcacttt attgtgaaga tagtggaaaa ggaaggtggc tcctacaaat
     3181 gccatcattg cgataaagga aaggccatcg ttgaagatgc ctctgccgac agtggtccca
     3241 aagatggacc cccacccacg aggagcatcg tggaaaaaga agacgttcca accacgtctt
     3301 caaagcaagt ggattgatgt gatatctcca ctgacgtaag ggatgacgca caatcccact
     3361 atccttcgca agacccttcc tctatataag gaagttcatt tcatttggag agaacacggg
     3421 ggactctaga ttgatacgaa acgtaccgta tcgttaaggt caccatggtg agtaaaggag
     3481 aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc
     3541 acaaattttc tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa cttaccctta
     3601 aatttatttg cactactgga aaactacctg ttccatggcc aaccctggtc accaccctga
     3661 cctacgtaag tttctgcttc tacctttgat atatatataa taattatcat taattagtag
     3721 taatataata tttcaaatat ttttttcaaa ataaaagaat gtagtatata gcaattgctt
     3781 ttctgtagtt tataagtgtg tatattttaa tttataactt ttctaatata tgaccaaaat
     3841 ttgttgatgt gcagggcgtg cagtgcttct cccgttaccc tgatcatatg aagcggcacg
     3901 acttcttcaa gagcgccatg cctgagggat acgtgcagga gaggaccatc ttcttcaagg
     3961 acgacgggaa ctacaagaca cgtgctgaag tcaagtttga gggtgacacc ctcgtcaaca
     4021 ggatcgagct taagggaatc gatttcaagg aggacggaaa catcctcggc cacaagttgg
     4081 aatacaacta caactcccac aacgtataca tcatggccga caagcaaaag aacggcatca
     4141 aagccaactt caagacccgc cacaacatcg aggacggcgg cgtgcaactc gctgatcatt
     4201 atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt accagacaac cattacctgt
     4261 ccacacaatc tgccctttcg aaagatccca acgaaaagag agatcacatg gtccttcttg
     4321 agtttgtaac agctgctggg attacacatg gcatggatga actatacaaa cctaagaaaa
     4381 agcgtaaggt cgaggaccct taatgtcaag ccattctctc gcagatgatg ttcactttgt
     4441 gttttacttc ctttatgcat tcacagcaat aaaagaaaga aatctccatc gcttttggtt
     4501 ttcttctctg tcttaagtta gtcgttttcg tgtctaatct attacttatc attgtaatag
     4561 actcttcttc tattgagatt tgaatataaa ctaaaacaca ttccatttta ctgtgttctc
     4621 aacattcaga atgcaaacgg actaaccgta gtgactcggc ttgagtcttc gggctagagc
     4681 ggactagttc tagagcggcc gccaccgcgg tggagctcca gcttttgttc cctttagtga
     4741 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4801 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4861 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4921 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4981 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5041 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5101 gcaggaaaga acatgaaggc cttgacagga tatattggcg ggtaaactaa gtcgctgtat
     5161 gtgtttgttt gagatctcat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5221 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5281 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5341 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5401 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5461 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5521 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5581 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5641 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgtgct
     5701 ctgctgaagc cagttacctt cggaagaaga gttggtagct cttgatccgg caaacaaacc
     5761 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5821 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5881 cgttaaggga ttttggtcat gaattcttgt tagacattat ttgccgacta ccttggtgat
     5941 ctcgcctttc acgtagtgaa caaattcttc caactgatct gcgcgcgagg ccaagcgatc
     6001 ttcttgtcca agataagcct gcctagcttc aagtatgacg ggctgatact gggccggcag
     6061 gcgctccatt gcccagtcgg cagcgacatc cttcggcgcg attttgccgg ttactgcgct
     6121 gtaccaaatg cgggacaacg taagcactac atttcgctca tcgccagccc agtcgggcgg
     6181 cgagttccat agcgttaagg tttcatttag cgcctcaaat agatcctgtt caggaaccgg
     6241 atcaaagagt tcctccgccg ctggacctac caaggcaacg ctatgttctc ttgcttttgt
     6301 cagcaagata gccagatcaa tgtcgatcgt ggctggctcg aagatacctg caagaatgtc
     6361 attgcgctgc cattctccaa attgcagttc gcgcttagct ggataacgcc acggaatgat
     6421 gtcgtcgtgc acaacaatgg tgacttctac agcgcggaga atctcgctct ctccagggga
     6481 agccgaagtt tccaaaaggt cgttgatcaa agctcgccgc gttgtttcat caagccttac
     6541 ggtcaccgta accagcaaat caatatcact gtgtggcttc aggccgccat ccactgcgga
     6601 gccgtacaaa tgtacggcca gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac
     6661 ctctgatagt tgagtcgata cttcggcgat caccgcttcc ctcatgatgt ttaactcctg
     6721 aattaagccg cgccgcgaag cggtgtcggc ttgaatgaat tgttaggcgt catcctgtgc
     6781 tcccgagaac cagtaccagt acatcgctgt ttcgttcgag acttgaggtc tagttttata
     6841 cgtgaacagg tcaatgccgc cgagagtaaa gccacatttt gcgtacaaat tgcaggcagg
     6901 tacattgttc gtttgtgcca tgatgatata tttttatctt gtgcaatgta acatcagaga
     6961 ttttgagaca caacgtggct ttgttgaata aatcgaactt ttgctgagtt gaaggatcag
     7021 atcacgcatc ttcccgacaa cgcagaccgt tccgtggcaa agcaaaagtt caaaatcacc
     7081 aactggtcca cctacaacaa agctctcatc aaccgtggct ccctcacttt ctggctggat
     7141 gatggggcga ttcaggagat ccccatccaa cagcccgccg tcgagcgggc ttttttatcc
     7201 ccggaagcct gtggatagag ggtagttatc cacgtgaaac cgctaatgcc ccgcaaagcc
     7261 ttgattcacg gggctttccg gcccgctcca aaaactatcc acgtgaaatc gctaatcagg
     7321 gtacgtgaaa tcgctaatcg gagtacgtga aatcgctaat aaggtcacgt gaaatcgcta
     7381 atcaaaaagg cacgtgagaa cgctaatagc cctttcagat caacagcttg caaacacccc
     7441 tcgctccggc aagtagttac agcaagtagt atgttcaatt agcttttcaa ttatgaatat
     7501 atatatcaat tattggtcgc ccttggcttg tggacaatgc gctacgcgca ccggctccgc
     7561 ccgtggacaa ccgcaagcgg ttgcccaccg tcgagcgcct ttgcccacaa cccggcggcc
     7621 ggccgcaaca gatcgtttta taaatttttt tttttgaaaa agaaaaagcc cgaaaggcgg
     7681 caacctctcg ggcttctgga tttc
//
