LOCUS       sequence-393619-        7095 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants 
            (proUBQ10::PIP2A-mCherry-NOSt_6XBetIO-min35S::GFPint-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7095)
  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 7095)
  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: JAB2512
FEATURES             Location/Qualifiers
     source          1..7095
                     /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        2768..2814
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     primer_bind     complement(2868..2896)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          3036..3224
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     3680..3700
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             3741..3761
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4101..4121)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4101..4119)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4140..4156)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4140..4156)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4153..4175)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4164..4180
                     /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(4188..4194,4195..4212,4213..4218))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4233..4254
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4371..4388)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4493..4517
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4608..5196)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4688..4707)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5287..6075)
                     /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      6533..6968
                     /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 taatgataga aaagttggct gcggccgcga
     2581 aattgattgg acgttcaata taaacattga ttggacgttc aatataataa ttgattggac
     2641 gttcaatata agaattgatt ggacgttcaa tataataatt gattggacgt tcaatataag
     2701 cattgattgg acgttcaata taacgggtct aagctagcta ggccacccac aatcccacta
     2761 tcctttcgca agacccttcc tctatataag gaagttcatt tcatttggag agaacacggg
     2821 ggactctaga caccatggtg agtaaaggag aagaactttt cactggagtt gtcccaattc
     2881 ttgttgaatt agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag
     2941 gtgatgcaac atacggaaaa cttaccctta aatttatttg cactactgga aaactacctg
     3001 ttccatggcc aaccctggtc accaccctga cctacgtaag tttctgcttc tacctttgat
     3061 atatatataa taattatcat taattagtag taatataata tttcaaatat ttttttcaaa
     3121 ataaaagaat gtagtatata gcaattgctt ttctgtagtt tataagtgtg tatattttaa
     3181 tttataactt ttctaatata tgaccaaaat ttgttgatgt gcagggcgtg cagtgcttct
     3241 cccgttaccc tgatcatatg aagcggcacg acttcttcaa gagcgccatg cctgagggat
     3301 acgtgcagga gaggaccatc ttcttcaagg acgacgggaa ctacaagaca cgtgctgaag
     3361 tcaagtttga gggtgacacc ctcgtcaaca ggatcgagct taagggaatc gatttcaagg
     3421 aggacggaaa catcctcggc cacaagttgg aatacaacta caactcccac aacgtataca
     3481 tcatggccga caagcaaaag aacggcatca aagccaactt caagacccgc cacaacatcg
     3541 aggacggcgg cgtgcaactc gctgatcatt atcaacaaaa tactccaatt ggcgatggcc
     3601 ctgtcctttt accagacaac cattacctgt ccacacaatc tgccctttcg aaagatccca
     3661 acgaaaagag agatcacatg gtccttcttg agtttgtaac agctgctggg attacacatg
     3721 gcatggatga actatacaaa cctaagaaaa agcgtaaggt cgaggaccct taatgtcaag
     3781 ccattctctc gcagatgatg ttcactttgt gttttacttc ctttatgcat tcacagcaat
     3841 aaaagaaaga aatctccatc gcttttggtt ttcttctctg tcttaagtta gtcgttttcg
     3901 tgtctaatct attacttatc attgtaatag actcttcttc tattgagatt tgaatataaa
     3961 ctaaaacaca ttccatttta ctgtgttctc aacattcaga atgcaaacgg actaaccgta
     4021 gtgactcggc ttgagtcttc gggctagagc ggactagttc tagagcggcc gccaccgcgg
     4081 tggagctcca gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg
     4141 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4201 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4261 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4321 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4381 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4441 atacggttat ccacagaatc aggggataac gcaggaaaga acatgaaggc cttgacagga
     4501 tatattggcg ggtaaactaa gtcgctgtat gtgtttgttt gagatctcat gtgagcaaaa
     4561 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4621 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4681 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4741 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4801 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4861 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4921 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4981 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5041 actagaagaa cagtatttgg tatctgtgct ctgctgaagc cagttacctt cggaagaaga
     5101 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5161 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5221 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gaattcttgt
     5281 tagacattat ttgccgacta ccttggtgat ctcgcctttc acgtagtgaa caaattcttc
     5341 caactgatct gcgcgcgagg ccaagcgatc ttcttgtcca agataagcct gcctagcttc
     5401 aagtatgacg ggctgatact gggccggcag gcgctccatt gcccagtcgg cagcgacatc
     5461 cttcggcgcg attttgccgg ttactgcgct gtaccaaatg cgggacaacg taagcactac
     5521 atttcgctca tcgccagccc agtcgggcgg cgagttccat agcgttaagg tttcatttag
     5581 cgcctcaaat agatcctgtt caggaaccgg atcaaagagt tcctccgccg ctggacctac
     5641 caaggcaacg ctatgttctc ttgcttttgt cagcaagata gccagatcaa tgtcgatcgt
     5701 ggctggctcg aagatacctg caagaatgtc attgcgctgc cattctccaa attgcagttc
     5761 gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc acaacaatgg tgacttctac
     5821 agcgcggaga atctcgctct ctccagggga agccgaagtt tccaaaaggt cgttgatcaa
     5881 agctcgccgc gttgtttcat caagccttac ggtcaccgta accagcaaat caatatcact
     5941 gtgtggcttc aggccgccat ccactgcgga gccgtacaaa tgtacggcca gcaacgtcgg
     6001 ttcgagatgg cgctcgatga cgccaactac ctctgatagt tgagtcgata cttcggcgat
     6061 caccgcttcc ctcatgatgt ttaactcctg aattaagccg cgccgcgaag cggtgtcggc
     6121 ttgaatgaat tgttaggcgt catcctgtgc tcccgagaac cagtaccagt acatcgctgt
     6181 ttcgttcgag acttgaggtc tagttttata cgtgaacagg tcaatgccgc cgagagtaaa
     6241 gccacatttt gcgtacaaat tgcaggcagg tacattgttc gtttgtgcca tgatgatata
     6301 tttttatctt gtgcaatgta acatcagaga ttttgagaca caacgtggct ttgttgaata
     6361 aatcgaactt ttgctgagtt gaaggatcag atcacgcatc ttcccgacaa cgcagaccgt
     6421 tccgtggcaa agcaaaagtt caaaatcacc aactggtcca cctacaacaa agctctcatc
     6481 aaccgtggct ccctcacttt ctggctggat gatggggcga ttcaggagat ccccatccaa
     6541 cagcccgccg tcgagcgggc ttttttatcc ccggaagcct gtggatagag ggtagttatc
     6601 cacgtgaaac cgctaatgcc ccgcaaagcc ttgattcacg gggctttccg gcccgctcca
     6661 aaaactatcc acgtgaaatc gctaatcagg gtacgtgaaa tcgctaatcg gagtacgtga
     6721 aatcgctaat aaggtcacgt gaaatcgcta atcaaaaagg cacgtgagaa cgctaatagc
     6781 cctttcagat caacagcttg caaacacccc tcgctccggc aagtagttac agcaagtagt
     6841 atgttcaatt agcttttcaa ttatgaatat atatatcaat tattggtcgc ccttggcttg
     6901 tggacaatgc gctacgcgca ccggctccgc ccgtggacaa ccgcaagcgg ttgcccaccg
     6961 tcgagcgcct ttgcccacaa cccggcggcc ggccgcaaca gatcgtttta taaatttttt
     7021 tttttgaaaa agaaaaagcc cgaaaggcgg caacctctcg ggcttctgga tttccgatcc
     7081 ccggaattag actct
//
