LOCUS       sequence-393424-        8509 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants 
            (proUBQ10::PIP2A-mCherry-NOSt_proG1090::AmtR-VP16-19St_4XAmtRO::GFPi
            nt-NLS-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8509)
  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 8509)
  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: JAB2602
FEATURES             Location/Qualifiers
     source          1..8509
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    13..35
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     CDS             1558..2268
                     /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(1703..1721)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      2287..2539
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        2730..2776
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     primer_bind     complement(2830..2858)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          2998..3186
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     3642..3662
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             3703..3723
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        4183..4229
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             4914..5147
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDTADFEFEQMFTDALGIDEYGG"
     CDS             5148..5168
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5224..5400
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(5486..5506)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5486..5504)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5525..5541)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5525..5541)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5538..5560)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5549..5565
                     /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(5573..5579,5580..5597,5598..5603))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5618..5639
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5756..5773)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5878..5902
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5993..6581)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6073..6092)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6668..7462)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      7959..8394
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 gaattagact cttggcagga tatattgtgg tgtaacgtta agctttggtt gtcgacgagt
       61 cagtaataaa cggcgtcaaa gtggttgcag ccggcacaca cgagtcgtgt ttatcaactc
      121 aaagcacaaa tacttttcct caacctaaaa ataaggcaat tagccaaaaa caactttgcg
      181 tgtaaacaac gctcaataca cgtgtcattt tattattagc tattgcttca ccgccttagc
      241 tttctcgtga cctagtcgtc ctcgtctttt cttcttcttc ttctataaaa caatacccaa
      301 agagctcttc ttcttcacaa ttcagatttc aatttctcaa aatcttaaaa actttctctc
      361 aattctctct accgtgatca aggtaaattt ctgtgttcct tattctctca aaatcttcga
      421 ttttgttttc gttcgatccc aatttcgtat atgttctttg gtttagattc tgttaatctt
      481 agatcgtaga cgattttctg ggtttgatcg ttagatatca tcttaattct cgattagggt
      541 ttcatagata tcatccgatt tgttcaaata atttgagttt tgtcgaataa ttactcttcg
      601 atttgtgatt tctatctaga tctggtgtta gtttctagtt tgtgcgatcg aatttgtcga
      661 ttaatctgag tttttctgat taacagcacc atggcaaagg atgtggaagc cgttcccgga
      721 gaaggatttc agacaagaga ctatcaagat ccgccaccag ctccgtttat tgatggagcg
      781 gagctaaaga agtggtcttt ctacagagca gttatcgcag agttcgtagc cactctcctc
      841 ttcttataca tcaccgtttt gacagtcatc ggttacaaga ttcagtccga tactgatgcc
      901 ggtggcgtag attgcggcgg agttggaatc ctcggtatcg cttgggcctt tggtggtatg
      961 atcttcatcc tcgtctactg caccgccggt atctctggtg gtcacattaa cccagcggtg
     1021 acatttgggc tattcttggc acgtaaagtg tcgttaccta gggccctatt gtacataatc
     1081 gctcagtgtt tgggtgcgat ttgtggagtt ggttttgtca aagccttcca aagctcttac
     1141 tacacccgtt acggaggtgg agccaactct ctagccgatg gctacagcac agggaccggt
     1201 ctagccgcag agatcattgg tactttcgtt cttgtctaca ccgtattctc tgccactgac
     1261 cccaaacgta gtgccagaga ctcccacgtt ccggtgttgg cgccacttcc aatcggattt
     1321 gccgtgttca tggtacattt ggctaccatt cccattaccg gaaccggaat taacccggca
     1381 aggagtttcg gagctgccgt aatctacaac aagagcaagc catgggatga ccactggata
     1441 ttttgggttg gaccattcat tggagctgcg atagctgcat tctaccacca attcgttcta
     1501 agagcttcag gttctaagtc tcttggatca ttcagaagtg ctgccaacgt cggatccatg
     1561 gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg
     1621 cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
     1681 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc
     1741 gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc
     1801 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
     1861 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
     1921 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
     1981 cagaataaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc
     2041 ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag
     2101 gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac
     2161 atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc
     2221 gccgagggcc gccactccac cggcggcatg gaggagctgt acaagtaaag cggccgcccg
     2281 gctgcagatc gttcaaacat ttggcaataa agtttcttaa gattgaatcc tgttgccggt
     2341 cttgcgatga ttatcatata atttctgttg aattacgtta agcatgtaat aattaacatg
     2401 taatgcatga cgttatttat gagatgggtt tttatgatta gagtcccgca attatacatt
     2461 taatacgcga tagaaaacaa aatatagcgc gcaaactagg ataaattatc gcgcgcggtg
     2521 tcatctatgt tactagatcg gggcgcgccc tcgagggtct ctaatgatag aaaagttggc
     2581 gcggccgcga atttctatcg atctatagat aatgttttct atcgatctat agataatgtt
     2641 ttctatcgat ctatagataa tagtttctat cgatctatag ataatcgggt ctaagctagc
     2701 taggccaccc acaatcccac tatcctttcg caagaccctt cctctatata aggaagttca
     2761 tttcatttgg agagaacacg ggggactcta gacaccatgg tgagtaaagg agaagaactt
     2821 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
     2881 tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt
     2941 tgcactactg gaaaactacc tgttccatgg ccaaccctgg tcaccaccct gacctacgta
     3001 agtttctgct tctacctttg atatatatat aataattatc attaattagt agtaatataa
     3061 tatttcaaat atttttttca aaataaaaga atgtagtata tagcaattgc ttttctgtag
     3121 tttataagtg tgtatatttt aatttataac ttttctaata tatgaccaaa atttgttgat
     3181 gtgcagggcg tgcagtgctt ctcccgttac cctgatcata tgaagcggca cgacttcttc
     3241 aagagcgcca tgcctgaggg atacgtgcag gagaggacca tcttcttcaa ggacgacggg
     3301 aactacaaga cacgtgctga agtcaagttt gagggtgaca ccctcgtcaa caggatcgag
     3361 cttaagggaa tcgatttcaa ggaggacgga aacatcctcg gccacaagtt ggaatacaac
     3421 tacaactccc acaacgtata catcatggcc gacaagcaaa agaacggcat caaagccaac
     3481 ttcaagaccc gccacaacat cgaggacggc ggcgtgcaac tcgctgatca ttatcaacaa
     3541 aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa
     3601 tctgcccttt cgaaagatcc caacgaaaag agagatcaca tggtccttct tgagtttgta
     3661 acagctgctg ggattacaca tggcatggat gaactataca aacctaagaa aaagcgtaag
     3721 gtcgaggacc cttaatgtca agccattctc tcgcagatga tgttcacttt gtgttttact
     3781 tcctttatgc attcacagca ataaaagaaa gaaatctcca tcgcttttgg ttttcttctc
     3841 tgtcttaagt tagtcgtttt cgtgtctaat ctattactta tcattgtaat agactcttct
     3901 tctattgaga tttgaatata aactaaaaca cattccattt tactgtgttc tcaacattca
     3961 gaatgcaaac ggactaaccg tagtgactcg gcttctaatg tagtttaaac tgaaggcggg
     4021 aaacgacaat ctgatccaag ctcaagctaa gcttgcatgc ctgcaggata tcgtggatcc
     4081 aagcttgcca cgtgccgcca cgtgccgcca cgtgccgcca cgtgcctcta gaggatccat
     4141 ctccactgac gtaagggatg acgcacaatc ccactatcct tcgcaagacc cttcctctat
     4201 ataaggaagt tcatttcatt tggagaggac acgctgggat ccccaccatg gctggcgccg
     4261 tgggcagacc cagaagatct gctcctcgga gagccggcaa gaacccccgg gaagagattc
     4321 tggatgccag cgccgagctg ttcaccagac agggctttgc caccaccagc acccaccaga
     4381 ttgccgacgc tgtgggcatc agacaggcca gcctgtacta ccacttcccc agcaagaccg
     4441 agatcttcct gaccctgctg aaaagcaccg tggaaccctc caccgtgctg gccgaggatc
     4501 tgtctaccct ggacgccgga cccgaaatga gactgtgggc tatcgtggcc agcgaagtgc
     4561 ggctgctgct gagcaccaag tggaacgtgg gccggctgta ccagctgccc atcgtgggct
     4621 ctgaggaatt cgccgagtac cacagccagc gcgaggccct gaccaacgtg ttcagagatc
     4681 tggccaccga gattgtgggc gacgacccca gagccgaact gcccttccac atcaccatga
     4741 gcgtgatcga gatgcggcgg aacgacggca agatccctag ccctctgagc gccgactctc
     4801 tgcccgagac agccattatg ctggctgacg cctccctggc tgtgctggga gcacctctgc
     4861 ctgccgacag agtggaaaag acactggaac tgatcaagca ggccgacgcc aaggcccccc
     4921 cgaccgacgt cagcctgggg gacgagctcc acttagacgg cgaggacgtg gcgatggcgc
     4981 atgccgacgc gctagacgat ttcgatctgg acatgttggg ggacggggat tccccggggc
     5041 cgggatttac cccccacgac tccgccccct acggcgctct ggatacggcc gacttcgagt
     5101 ttgagcagat gtttaccgat gcccttggaa ttgacgagta cggtgggccg aagaagaaga
     5161 ggaaggttta gtgtcggtac gctgaaatca ccagtctctc tctacaaatc tatctctctc
     5221 tattttctcc ataaataatg tgtgagtagt ttcccgataa gggaaattag ggttcttata
     5281 gggtttcgct catgtgttga gcatataaga aacccttagt atgtatttgt atttgtaaaa
     5341 tacttctatc aataaaattt ctaattccta aaaccaaaat ccagtactaa aatccagatc
     5401 gatgcttgat actcgcttcg agaccgggct agagcggact agttctagag cggccgccac
     5461 cgcggtggag ctccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat
     5521 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5581 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5641 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5701 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5761 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5821 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg aaggccttga
     5881 caggatatat tggcgggtaa actaagtcgc tgtatgtgtt tgtttgagat ctcatgtgag
     5941 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6001 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6061 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6121 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6181 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6241 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6301 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6361 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6421 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6481 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6541 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6601 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgcatt
     6661 ctaggtacta aaacaattca tccagtaaaa tataatattt tattttctcc caatcaggct
     6721 tgatccccag taagtcaaaa aatagctcga catactgttc ttccccgata tcctccctga
     6781 tcgaccggac gcagaaggca atgtcatacc acttgtccgc cctgccgctt ctcccaagat
     6841 caataaagcc acttactttg ccatctttca caaagatgtt gctgtcaccc aggtcgccgt
     6901 gggaaaagac aagttcctct tcgggctttt ccgtctttaa aaaatcatac agctcgcgcg
     6961 gatctttaaa tggagtatct tcttcccagt tttcgcaatc cacatcggcc agatcgttat
     7021 tcagtaagta atccaattcg gctaagcggc tgtctaagct attcgtatag ggacaatccg
     7081 atatgtcgat ggagtgaaag agcctgatgc actccgcata cagctcgata atcttttcag
     7141 ggctttgttc atcttcatac tcttccgagc aaaggacgcc atcggcctca ctcatgagca
     7201 gattgctcca gccatcatgc cgttcaaagt gcaggacctt tggaacaggc agctttcctt
     7261 ccagccatag catcatgtcc ttttcccgtt ccacatcata ggtggtccct ttataccggc
     7321 tgtccgtcat ttttaaatat aggttttcat tttctcccac cagcttatat accttagcag
     7381 gtgacattcc ttccgtatct tttacgcagc ggtatttttc gatcagtttt ttcaattccg
     7441 gtgatattct cattttagcc atttattatt tccttcctct tttctacagt atttaaagat
     7501 accccaagaa gctaattata acaagacgaa ctccaattca ctgttccttg cattctaaaa
     7561 ccttaaatac cagaaaacag ctttttcaaa gttgttttca aagttggcgt ataacatagt
     7621 atcgacggag ccgattttga aaccgcggtg atcacaggca gcaacgctct gtcatcgtta
     7681 caatcaacat gctaccctcc gcgagatcat ccgtgtttca aacccggcag cttattgtgc
     7741 aatgtaacat cagagatttt gagacacaac gtggctttgt tgaataaatc gaacttttgc
     7801 tgagttgaag gatcagatca cgcatcttcc cgacaacgca gaccgttccg tggcaaagca
     7861 aaagttcaaa atcaccaact ggtccaccta caacaaagct ctcatcaacc gtggctccct
     7921 cactttctgg ctggatgatg gggcgattca ggagatcccc atccaacagc ccgccgtcga
     7981 gcgggctttt ttatccccgg aagcctgtgg atagagggta gttatccacg tgaaaccgct
     8041 aatgccccgc aaagccttga ttcacggggc tttccggccc gctccaaaaa ctatccacgt
     8101 gaaatcgcta atcagggtac gtgaaatcgc taatcggagt acgtgaaatc gctaataagg
     8161 tcacgtgaaa tcgctaatca aaaaggcacg tgagaacgct aatagccctt tcagatcaac
     8221 agcttgcaaa cacccctcgc tccggcaagt agttacagca agtagtatgt tcaattagct
     8281 tttcaattat gaatatatat atcaattatt ggtcgccctt ggcttgtgga caatgcgcta
     8341 cgcgcaccgg ctccgcccgt ggacaaccgc aagcggttgc ccaccgtcga gcgcctttgc
     8401 ccacaacccg gcggccggcc gcaacagatc gttttataaa tttttttttt tgaaaaagaa
     8461 aaagcccgaa aggcggcaac ctctcgggct tctggatttc cgatccccg
//
