LOCUS       sequence-393418-        7035 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants 
            (proUBQ10::PIP2A-mCherry-NOSt_6XMcbRO-min35S::GFPint-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7035)
  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 7035)
  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: JAB2185
FEATURES             Location/Qualifiers
     source          1..7035
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    80..102
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     CDS             1625..2335
                     /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(1770..1788)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      2354..2606
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        2817..2863
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     primer_bind     complement(2917..2945)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          3085..3273
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     3729..3749
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4120..4140)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4120..4138)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4159..4175)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4159..4175)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4172..4194)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4183..4199
                     /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(4207..4213,4214..4231,4232..4237))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4252..4273
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4390..4407)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4512..4536
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4627..5215)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4707..4726)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5306..6094)
                     /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      6552..6987
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 tttttttttg aaaaagaaaa agcccgaaag gcggcaacct ctcgggcttc tggatttccg
       61 atccccggaa ttagactctt ggcaggatat attgtggtgt aacgttaagc tttggttgtc
      121 gacgagtcag taataaacgg cgtcaaagtg gttgcagccg gcacacacga gtcgtgttta
      181 tcaactcaaa gcacaaatac ttttcctcaa cctaaaaata aggcaattag ccaaaaacaa
      241 ctttgcgtgt aaacaacgct caatacacgt gtcattttat tattagctat tgcttcaccg
      301 ccttagcttt ctcgtgacct agtcgtcctc gtcttttctt cttcttcttc tataaaacaa
      361 tacccaaaga gctcttcttc ttcacaattc agatttcaat ttctcaaaat cttaaaaact
      421 ttctctcaat tctctctacc gtgatcaagg taaatttctg tgttccttat tctctcaaaa
      481 tcttcgattt tgttttcgtt cgatcccaat ttcgtatatg ttctttggtt tagattctgt
      541 taatcttaga tcgtagacga ttttctgggt ttgatcgtta gatatcatct taattctcga
      601 ttagggtttc atagatatca tccgatttgt tcaaataatt tgagttttgt cgaataatta
      661 ctcttcgatt tgtgatttct atctagatct ggtgttagtt tctagtttgt gcgatcgaat
      721 ttgtcgatta atctgagttt ttctgattaa cagcaccatg gcaaaggatg tggaagccgt
      781 tcccggagaa ggatttcaga caagagacta tcaagatccg ccaccagctc cgtttattga
      841 tggagcggag ctaaagaagt ggtctttcta cagagcagtt atcgcagagt tcgtagccac
      901 tctcctcttc ttatacatca ccgttttgac agtcatcggt tacaagattc agtccgatac
      961 tgatgccggt ggcgtagatt gcggcggagt tggaatcctc ggtatcgctt gggcctttgg
     1021 tggtatgatc ttcatcctcg tctactgcac cgccggtatc tctggtggtc acattaaccc
     1081 agcggtgaca tttgggctat tcttggcacg taaagtgtcg ttacctaggg ccctattgta
     1141 cataatcgct cagtgtttgg gtgcgatttg tggagttggt tttgtcaaag ccttccaaag
     1201 ctcttactac acccgttacg gaggtggagc caactctcta gccgatggct acagcacagg
     1261 gaccggtcta gccgcagaga tcattggtac tttcgttctt gtctacaccg tattctctgc
     1321 cactgacccc aaacgtagtg ccagagactc ccacgttccg gtgttggcgc cacttccaat
     1381 cggatttgcc gtgttcatgg tacatttggc taccattccc attaccggaa ccggaattaa
     1441 cccggcaagg agtttcggag ctgccgtaat ctacaacaag agcaagccat gggatgacca
     1501 ctggatattt tgggttggac cattcattgg agctgcgata gctgcattct accaccaatt
     1561 cgttctaaga gcttcaggtt ctaagtctct tggatcattc agaagtgctg ccaacgtcgg
     1621 atccatggtg agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt
     1681 caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga
     1741 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct
     1801 gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa
     1861 gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga
     1921 gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca
     1981 ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc
     2041 cgtaatgcag aataagacca tgggctggga ggcctcctcc gagcggatgt accccgagga
     2101 cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga
     2161 cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa
     2221 cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta
     2281 cgaacgcgcc gagggccgcc actccaccgg cggcatggag gagctgtaca agtaaagcgg
     2341 ccgcccggct gcagatcgtt caaacatttg gcaataaagt ttcttaagat tgaatcctgt
     2401 tgccggtctt gcgatgatta tcatataatt tctgttgaat tacgttaagc atgtaataat
     2461 taacatgtaa tgcatgacgt tatttatgag atgggttttt atgattagag tcccgcaatt
     2521 atacatttaa tacgcgatag aaaacaaaat atagcgcgca aactaggata aattatcgcg
     2581 cgcggtgtca tctatgttac tagatcgggg cgcgccctcg aggaagacgt aatgatagaa
     2641 aagttggctg cggccgcgaa atagactggc ctgtctagaa tagactggcc tgtctacaat
     2701 agactggcct gtctacaata gactggcctg tctagtatag actggcctgt ctaagataga
     2761 ctggcctgtc tacgggtcta agctagctag gccacccaca atcccactat cctttcgcaa
     2821 gacccttcct ctatataagg aagttcattt catttggaga gaacacgggg gactctagac
     2881 accatggtga gtaaaggaga agaacttttc actggagttg tcccaattct tgttgaatta
     2941 gatggtgatg ttaatgggca caaattttct gtcagtggag agggtgaagg tgatgcaaca
     3001 tacggaaaac ttacccttaa atttatttgc actactggaa aactacctgt tccatggcca
     3061 accctggtca ccaccctgac ctacgtaagt ttctgcttct acctttgata tatatataat
     3121 aattatcatt aattagtagt aatataatat ttcaaatatt tttttcaaaa taaaagaatg
     3181 tagtatatag caattgcttt tctgtagttt ataagtgtgt atattttaat ttataacttt
     3241 tctaatatat gaccaaaatt tgttgatgtg cagggcgtgc agtgcttctc ccgttaccct
     3301 gatcatatga agcggcacga cttcttcaag agcgccatgc ctgagggata cgtgcaggag
     3361 aggaccatct tcttcaagga cgacgggaac tacaagacac gtgctgaagt caagtttgag
     3421 ggtgacaccc tcgtcaacag gatcgagctt aagggaatcg atttcaagga ggacggaaac
     3481 atcctcggcc acaagttgga atacaactac aactcccaca acgtatacat catggccgac
     3541 aagcaaaaga acggcatcaa agccaacttc aagacccgcc acaacatcga ggacggcggc
     3601 gtgcaactcg ctgatcatta tcaacaaaat actccaattg gcgatggccc tgtcctttta
     3661 ccagacaacc attacctgtc cacacaatct gccctttcga aagatcccaa cgaaaagaga
     3721 gatcacatgg tccttcttga gtttgtaaca gctgctggga ttacacatgg catggatgaa
     3781 ctatacaaat aatgtcaagc cattctctcg cagatgatgt tcactttgtg ttttacttcc
     3841 tttatgcatt cacagcaata aaagaaagaa atctccatcg cttttggttt tcttctctgt
     3901 cttaagttag tcgttttcgt gtctaatcta ttacttatca ttgtaataga ctcttcttct
     3961 attgagattt gaatataaac taaaacacat tccattttac tgtgttctca acattcagaa
     4021 tgcaaacgga ctaaccgtag tgactcggct tgagtcttcg ggctagagcg gactagttct
     4081 agagcggccg ccaccgcggt ggagctccag cttttgttcc ctttagtgag ggttaattgc
     4141 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4201 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4261 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4321 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4381 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4441 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4501 catgaaggcc ttgacaggat atattggcgg gtaaactaag tcgctgtatg tgtttgtttg
     4561 agatctcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4621 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4681 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4741 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4801 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4861 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4921 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4981 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5041 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgtgctc tgctgaagcc
     5101 agttaccttc ggaagaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5161 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5221 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5281 tttggtcatg aattcttgtt agacattatt tgccgactac cttggtgatc tcgcctttca
     5341 cgtagtgaac aaattcttcc aactgatctg cgcgcgaggc caagcgatct tcttgtccaa
     5401 gataagcctg cctagcttca agtatgacgg gctgatactg ggccggcagg cgctccattg
     5461 cccagtcggc agcgacatcc ttcggcgcga ttttgccggt tactgcgctg taccaaatgc
     5521 gggacaacgt aagcactaca tttcgctcat cgccagccca gtcgggcggc gagttccata
     5581 gcgttaaggt ttcatttagc gcctcaaata gatcctgttc aggaaccgga tcaaagagtt
     5641 cctccgccgc tggacctacc aaggcaacgc tatgttctct tgcttttgtc agcaagatag
     5701 ccagatcaat gtcgatcgtg gctggctcga agatacctgc aagaatgtca ttgcgctgcc
     5761 attctccaaa ttgcagttcg cgcttagctg gataacgcca cggaatgatg tcgtcgtgca
     5821 caacaatggt gacttctaca gcgcggagaa tctcgctctc tccaggggaa gccgaagttt
     5881 ccaaaaggtc gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa
     5941 ccagcaaatc aatatcactg tgtggcttca ggccgccatc cactgcggag ccgtacaaat
     6001 gtacggccag caacgtcggt tcgagatggc gctcgatgac gccaactacc tctgatagtt
     6061 gagtcgatac ttcggcgatc accgcttccc tcatgatgtt taactcctga attaagccgc
     6121 gccgcgaagc ggtgtcggct tgaatgaatt gttaggcgtc atcctgtgct cccgagaacc
     6181 agtaccagta catcgctgtt tcgttcgaga cttgaggtct agttttatac gtgaacaggt
     6241 caatgccgcc gagagtaaag ccacattttg cgtacaaatt gcaggcaggt acattgttcg
     6301 tttgtgccat gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac
     6361 aacgtggctt tgttgaataa atcgaacttt tgctgagttg aaggatcaga tcacgcatct
     6421 tcccgacaac gcagaccgtt ccgtggcaaa gcaaaagttc aaaatcacca actggtccac
     6481 ctacaacaaa gctctcatca accgtggctc cctcactttc tggctggatg atggggcgat
     6541 tcaggagatc cccatccaac agcccgccgt cgagcgggct tttttatccc cggaagcctg
     6601 tggatagagg gtagttatcc acgtgaaacc gctaatgccc cgcaaagcct tgattcacgg
     6661 ggctttccgg cccgctccaa aaactatcca cgtgaaatcg ctaatcaggg tacgtgaaat
     6721 cgctaatcgg agtacgtgaa atcgctaata aggtcacgtg aaatcgctaa tcaaaaaggc
     6781 acgtgagaac gctaatagcc ctttcagatc aacagcttgc aaacacccct cgctccggca
     6841 agtagttaca gcaagtagta tgttcaatta gcttttcaat tatgaatata tatatcaatt
     6901 attggtcgcc cttggcttgt ggacaatgcg ctacgcgcac cggctccgcc cgtggacaac
     6961 cgcaagcggt tgcccaccgt cgagcgcctt tgcccacaac ccggcggccg gccgcaacag
     7021 atcgttttat aaatt
//
