LOCUS       sequence-391161-        8462 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_2XAmtRO::GFPi
            nt-NLS-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8462)
  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 8462)
  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: JAB2600
FEATURES             Location/Qualifiers
     source          1..8462
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1541..2251
                     /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(1686..1704)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      2270..2522
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        2666..2712
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     primer_bind     complement(2766..2794)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          2934..3122
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     3578..3598
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             3639..3659
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        4119..4165
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             4850..5083
                     /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             5084..5104
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5160..5336
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(5422..5442)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5422..5440)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5461..5477)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5461..5477)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5474..5496)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5485..5501
                     /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(5509..5515,5516..5533,5534..5539))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5554..5575
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5692..5709)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5814..5838
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5929..6517)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6009..6028)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6604..7398)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      7895..8330
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(8458..8462,1..18)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
ORIGIN
        1 ggatatattg tggtgtaacg ttaagctttg gttgtcgacg agtcagtaat aaacggcgtc
       61 aaagtggttg cagccggcac acacgagtcg tgtttatcaa ctcaaagcac aaatactttt
      121 cctcaaccta aaaataaggc aattagccaa aaacaacttt gcgtgtaaac aacgctcaat
      181 acacgtgtca ttttattatt agctattgct tcaccgcctt agctttctcg tgacctagtc
      241 gtcctcgtct tttcttcttc ttcttctata aaacaatacc caaagagctc ttcttcttca
      301 caattcagat ttcaatttct caaaatctta aaaactttct ctcaattctc tctaccgtga
      361 tcaaggtaaa tttctgtgtt ccttattctc tcaaaatctt cgattttgtt ttcgttcgat
      421 cccaatttcg tatatgttct ttggtttaga ttctgttaat cttagatcgt agacgatttt
      481 ctgggtttga tcgttagata tcatcttaat tctcgattag ggtttcatag atatcatccg
      541 atttgttcaa ataatttgag ttttgtcgaa taattactct tcgatttgtg atttctatct
      601 agatctggtg ttagtttcta gtttgtgcga tcgaatttgt cgattaatct gagtttttct
      661 gattaacagc accatggcaa aggatgtgga agccgttccc ggagaaggat ttcagacaag
      721 agactatcaa gatccgccac cagctccgtt tattgatgga gcggagctaa agaagtggtc
      781 tttctacaga gcagttatcg cagagttcgt agccactctc ctcttcttat acatcaccgt
      841 tttgacagtc atcggttaca agattcagtc cgatactgat gccggtggcg tagattgcgg
      901 cggagttgga atcctcggta tcgcttgggc ctttggtggt atgatcttca tcctcgtcta
      961 ctgcaccgcc ggtatctctg gtggtcacat taacccagcg gtgacatttg ggctattctt
     1021 ggcacgtaaa gtgtcgttac ctagggccct attgtacata atcgctcagt gtttgggtgc
     1081 gatttgtgga gttggttttg tcaaagcctt ccaaagctct tactacaccc gttacggagg
     1141 tggagccaac tctctagccg atggctacag cacagggacc ggtctagccg cagagatcat
     1201 tggtactttc gttcttgtct acaccgtatt ctctgccact gaccccaaac gtagtgccag
     1261 agactcccac gttccggtgt tggcgccact tccaatcgga tttgccgtgt tcatggtaca
     1321 tttggctacc attcccatta ccggaaccgg aattaacccg gcaaggagtt tcggagctgc
     1381 cgtaatctac aacaagagca agccatggga tgaccactgg atattttggg ttggaccatt
     1441 cattggagct gcgatagctg cattctacca ccaattcgtt ctaagagctt caggttctaa
     1501 gtctcttgga tcattcagaa gtgctgccaa cgtcggatcc atggtgagca agggcgagga
     1561 ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt
     1621 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
     1681 gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc
     1741 ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta
     1801 cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
     1861 cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt
     1921 gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaata agaccatggg
     1981 ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa
     2041 gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa
     2101 ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac
     2161 ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc
     2221 caccggcggc atggaggagc tgtacaagta aagcggccgc ccggctgcag atcgttcaaa
     2281 catttggcaa taaagtttct taagattgaa tcctgttgcc ggtcttgcga tgattatcat
     2341 ataatttctg ttgaattacg ttaagcatgt aataattaac atgtaatgca tgacgttatt
     2401 tatgagatgg gtttttatga ttagagtccc gcaattatac atttaatacg cgatagaaaa
     2461 caaaatatag cgcgcaaact aggataaatt atcgcgcgcg gtgtcatcta tgttactaga
     2521 tcggggcgcg ccctcgaggg tctctaatga tagaaaagtt ggctgcggcc gcgaatttct
     2581 atcgatctat agataatagt ttctatcgat ctatagataa tcgggtctaa gctagctagg
     2641 ccacccacaa tcccactatc ctttcgcaag acccttcctc tatataagga agttcatttc
     2701 atttggagag aacacggggg actctagaca ccatggtgag taaaggagaa gaacttttca
     2761 ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg
     2821 tcagtggaga gggtgaaggt gatgcaacat acggaaaact tacccttaaa tttatttgca
     2881 ctactggaaa actacctgtt ccatggccaa ccctggtcac caccctgacc tacgtaagtt
     2941 tctgcttcta cctttgatat atatataata attatcatta attagtagta atataatnnn
     3001 tcaaatattt ttttcaaaat aaaagaatgt agtatatagc aattgctttt ctgtagttta
     3061 taagtgtgta tattttaatt tataactttt ctaatatatg accaaaattt gttgatgtgc
     3121 agggcgtgca gtgcttctcc cgttaccctg atcatatgaa gcggcacgac ttcttcaaga
     3181 gcgccatgcc tgagggatac gtgcaggaga ggaccatctt cttcaaggac gacgggaact
     3241 acaagacacg tgctgaagtc aagtttgagg gtgacaccct cgtcaacagg atcgagctta
     3301 agggaatcga tttcaaggag gacggaaaca tcctcggcca caagttggaa tacaactaca
     3361 actcccacaa cgtatacatc atggccgaca agcaaaagaa cggcatcaaa gccaacttca
     3421 agacccgcca caacatcgag gacggcggcg tgcaactcgc tgatcattat caacaaaata
     3481 ctccaattgg cgatggccct gtccttttac cagacaacca ttacctgtcc acacaatctg
     3541 ccctttcgaa agatcccaac gaaaagagag atcacatggt ccttcttgag tttgtaacag
     3601 ctgctgggat tacacatggc atggatgaac tatacaaacc taagaaaaag cgtaaggtcg
     3661 aggaccctta atgtcaagcc attctctcgc agatgatgtt cactttgtgt tttacttcct
     3721 ttatgcattc acagcaataa aagaaagaaa tctccatcgc ttttggtttt cttctctgtc
     3781 ttaagttagt cgttttcgtg tctaatctat tacttatcat tgtaatagac tcttcttcta
     3841 ttgagatttg aatataaact aaaacacatt ccattttact gtgttctcaa cattcagaat
     3901 gcaaacggac taaccgtagt gactcggctt ctaatgtagt ttaaactgaa ggcgggaaac
     3961 gacaatctga tccaagctca agctaagctt gcatgcctgc aggatatcgt ggatccaagc
     4021 ttgccacgtg ccgccacgtg ccgccacgtg ccgccacgtg cctctagagg atccatctcc
     4081 actgacgtaa gggatgacgc acaatcccac tatccttcgc aagacccttc ctctatataa
     4141 ggaagttcat ttcatttgga gaggacacgc tgggatcccc accatggctg gcgccgtggg
     4201 cagacccaga agatctgctc ctcggagagc cggcaagaac ccccgggaag agattctgga
     4261 tgccagcgcc gagctgttca ccagacaggg ctttgccacc accagcaccc accagattgc
     4321 cgacgctgtg ggcatcagac aggccagcct gtactaccac ttccccagca agaccgagat
     4381 cttcctgacc ctgctgaaaa gcaccgtgga accctccacc gtgctggccg aggatctgtc
     4441 taccctggac gccggacccg aaatgagact gtgggctatc gtggccagcg aagtgcggct
     4501 gctgctgagc accaagtgga acgtgggccg gctgtaccag ctgcccatcg tgggctctga
     4561 ggaattcgcc gagtaccaca gccagcgcga ggccctgacc aacgtgttca gagatctggc
     4621 caccgagatt gtgggcgacg accccagagc cgaactgccc ttccacatca ccatgagcgt
     4681 gatcgagatg cggcggaacg acggcaagat ccctagccct ctgagcgccg actctctgcc
     4741 cgagacagcc attatgctgg ctgacgcctc cctggctgtg ctgggagcac ctctgcctgc
     4801 cgacagagtg gaaaagacac tggaactgat caagcaggcc gacgccaagg cccccccgac
     4861 cgacgtcagc ctgggggacg agctccactt agacggcgag gacgtggcga tggcgcatgc
     4921 cgacgcgcta gacgatttcg atctggacat gttgggggac ggggattccc cggggccggg
     4981 atttaccccc cacgactccg ccccctacgg cgctctggat acggccgact tcgagtttga
     5041 gcagatgttt accgatgccc ttggaattga cgagtacggt gggccgaaga agaagaggaa
     5101 ggtttagtgt cggtacgctg aaatcaccag tctctctcta caaatctatc tctctctatt
     5161 ttctccataa ataatgtgtg agtagtttcc cgataaggga aattagggtt cttatagggt
     5221 ttcgctcatg tgttgagcat ataagaaacc cttagtatgt atttgtattt gtaaaatact
     5281 tctatcaata aaatttctaa ttcctaaaac caaaatccag tactaaaatc cagatcgatg
     5341 cttgatactc gcttcgagac cgggctagag cggactagtt ctagagcggc cgccaccgcg
     5401 gtggagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg
     5461 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5521 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5581 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5641 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5701 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5761 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgaagg ccttgacagg
     5821 atatattggc gggtaaacta agtcgctgta tgtgtttgtt tgagatctca tgtgagcaaa
     5881 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5941 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6001 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6061 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6121 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6181 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6241 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6301 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6361 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6421 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6481 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6541 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgcattctag
     6601 gtactaaaac aattcatcca gtaaaatata atattttatt ttctcccaat caggcttgat
     6661 ccccagtaag tcaaaaaata gctcgacata ctgttcttcc ccgatatcct ccctgatcga
     6721 ccggacgcag aaggcaatgt cataccactt gtccgccctg ccgcttctcc caagatcaat
     6781 aaagccactt actttgccat ctttcacaaa gatgttgctg tcacccaggt cgccgtggga
     6841 aaagacaagt tcctcttcgg gcttttccgt ctttaaaaaa tcatacagct cgcgcggatc
     6901 tttaaatgga gtatcttctt cccagttttc gcaatccaca tcggccagat cgttattcag
     6961 taagtaatcc aattcggcta agcggctgtc taagctattc gtatagggac aatccgatat
     7021 gtcgatggag tgaaagagcc tgatgcactc cgcatacagc tcgataatct tttcagggct
     7081 ttgttcatct tcatactctt ccgagcaaag gacgccatcg gcctcactca tgagcagatt
     7141 gctccagcca tcatgccgtt caaagtgcag gacctttgga acaggcagct ttccttccag
     7201 ccatagcatc atgtcctttt cccgttccac atcataggtg gtccctttat accggctgtc
     7261 cgtcattttt aaatataggt tttcattttc tcccaccagc ttatatacct tagcaggtga
     7321 cattccttcc gtatctttta cgcagcggta tttttcgatc agttttttca attccggtga
     7381 tattctcatt ttagccattt attatttcct tcctcttttc tacagtattt aaagataccc
     7441 caagaagcta attataacaa gacgaactcc aattcactgt tccttgcatt ctaaaacctt
     7501 aaataccaga aaacagcttt ttcaaagttg ttttcaaagt tggcgtataa catagtatcg
     7561 acggagccga ttttgaaacc gcggtgatca caggcagcaa cgctctgtca tcgttacaat
     7621 caacatgcta ccctccgcga gatcatccgt gtttcaaacc cggcagctta ttgtgcaatg
     7681 taacatcaga gattttgaga cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag
     7741 ttgaaggatc agatcacgca tcttcccgac aacgcagacc gttccgtggc aaagcaaaag
     7801 ttcaaaatca ccaactggtc cacctacaac aaagctctca tcaaccgtgg ctccctcact
     7861 ttctggctgg atgatggggc gattcaggag atccccatcc aacagcccgc cgtcgagcgg
     7921 gcttttttat ccccggaagc ctgtggatag agggtagtta tccacgtgaa accgctaatg
     7981 ccccgcaaag ccttgattca cggggctttc cggcccgctc caaaaactat ccacgtgaaa
     8041 tcgctaatca gggtacgtga aatcgctaat cggagtacgt gaaatcgcta ataaggtcac
     8101 gtgaaatcgc taatcaaaaa ggcacgtgag aacgctaata gccctttcag atcaacagct
     8161 tgcaaacacc cctcgctccg gcaagtagtt acagcaagta gtatgttcaa ttagcttttc
     8221 aattatgaat atatatatca attattggtc gcccttggct tgtggacaat gcgctacgcg
     8281 caccggctcc gcccgtggac aaccgcaagc ggttgcccac cgtcgagcgc ctttgcccac
     8341 aacccggcgg ccggccgcaa cagatcgttt tataaatttt tttttttgaa aaagaaaaag
     8401 cccgaaaggc ggcaacctct cgggcttctg gatttccgat ccccggaatt agactcttgg
     8461 ca
//
