LOCUS       sequence-389752-        8438 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_1XAmtRO::GFPi
            nt-NLS-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8438)
  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 8438)
  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: JAB2599
FEATURES             Location/Qualifiers
     source          1..8438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      98..533
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     CDS             1030..1824
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      1911..2499
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2400..2419
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2590..2614
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     2719..2736
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(2889..2894,2895..2912,2913..2919)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2927..2943
                     /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)."
     primer_bind     2932..2954
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2951..2967
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2951..2967
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2986..3006
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2988..3006
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    3092..3268
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(3324..3344)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(3345..3578)
                     /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"
     promoter        complement(4263..4309)
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             complement(4769..4789)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4830..4850)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     intron          5306..5494
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     5634..5662
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     promoter        complement(5716..5762)
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     terminator      complement(5882..6134)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(6153..6863)
                     /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     6700..6718
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     misc_feature    8386..8408
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
ORIGIN
        1 aagcccgaga ggttgccgcc tttcgggctt tttctttttc aaaaaaaaaa atttataaaa
       61 cgatctgttg cggccggccg ccgggttgtg ggcaaaggcg ctcgacggtg ggcaaccgct
      121 tgcggttgtc cacgggcgga gccggtgcgc gtagcgcatt gtccacaagc caagggcgac
      181 caataattga tatatatatt cataattgaa aagctaattg aacatactac ttgctgtaac
      241 tacttgccgg agcgaggggt gtttgcaagc tgttgatctg aaagggctat tagcgttctc
      301 acgtgccttt ttgattagcg atttcacgtg accttattag cgatttcacg tactccgatt
      361 agcgatttca cgtaccctga ttagcgattt cacgtggata gtttttggag cgggccggaa
      421 agccccgtga atcaaggctt tgcggggcat tagcggtttc acgtggataa ctaccctcta
      481 tccacaggct tccggggata aaaaagcccg ctcgacggcg ggctgttgga tggggatctc
      541 ctgaatcgcc ccatcatcca gccagaaagt gagggagcca cggttgatga gagctttgtt
      601 gtaggtggac cagttggtga ttttgaactt ttgctttgcc acggaacggt ctgcgttgtc
      661 gggaagatgc gtgatctgat ccttcaactc agcaaaagtt cgatttattc aacaaagcca
      721 cgttgtgtct caaaatctct gatgttacat tgcacaataa gctgccgggt ttgaaacacg
      781 gatgatctcg cggagggtag catgttgatt gtaacgatga cagagcgttg ctgcctgtga
      841 tcaccgcggt ttcaaaatcg gctccgtcga tactatgtta tacgccaact ttgaaaacaa
      901 ctttgaaaaa gctgttttct ggtatttaag gttttagaat gcaaggaaca gtgaattgga
      961 gttcgtcttg ttataattag cttcttgggg tatctttaaa tactgtagaa aagaggaagg
     1021 aaataataaa tggctaaaat gagaatatca ccggaattga aaaaactgat cgaaaaatac
     1081 cgctgcgtaa aagatacgga aggaatgtca cctgctaagg tatataagct ggtgggagaa
     1141 aatgaaaacc tatatttaaa aatgacggac agccggtata aagggaccac ctatgatgtg
     1201 gaacgggaaa aggacatgat gctatggctg gaaggaaagc tgcctgttcc aaaggtcctg
     1261 cactttgaac ggcatgatgg ctggagcaat ctgctcatga gtgaggccga tggcgtcctt
     1321 tgctcggaag agtatgaaga tgaacaaagc cctgaaaaga ttatcgagct gtatgcggag
     1381 tgcatcaggc tctttcactc catcgacata tcggattgtc cctatacgaa tagcttagac
     1441 agccgcttag ccgaattgga ttacttactg aataacgatc tggccgatgt ggattgcgaa
     1501 aactgggaag aagatactcc atttaaagat ccgcgcgagc tgtatgattt tttaaagacg
     1561 gaaaagcccg aagaggaact tgtcttttcc cacggcgacc tgggtgacag caacatcttt
     1621 gtgaaagatg gcaaagtaag tggctttatt gatcttggga gaagcggcag ggcggacaag
     1681 tggtatgaca ttgccttctg cgtccggtcg atcagggagg atatcgggga agaacagtat
     1741 gtcgagctat tttttgactt actggggatc aagcctgatt gggagaaaat aaaatattat
     1801 attttactgg atgaattgtt ttagtaccta gaatgcatga ccaaaatccc ttaacgtgag
     1861 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     1921 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     1981 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2041 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     2101 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2161 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2221 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2281 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     2341 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     2401 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     2461 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     2521 ttacggttcc tggccttttg ctggcctttt gctcacatga gatctcaaac aaacacatac
     2581 agcgacttag tttacccgcc aatatatcct gtcaaggcct tcatgttctt tcctgcgtta
     2641 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     2701 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     2761 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     2821 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcgc tcattaggca
     2881 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     2941 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac
     3001 taaagggaac aaaagctgga gctccaccgc ggtggcggcc gctctagaac tagtccgctc
     3061 tagcccggtc tcgaagcgag tatcaagcat cgatctggat tttagtactg gattttggtt
     3121 ttaggaatta gaaattttat tgatagaagt attttacaaa tacaaataca tactaagggt
     3181 ttcttatatg ctcaacacat gagcgaaacc ctataagaac cctaatttcc cttatcggga
     3241 aactactcac acattattta tggagaaaat agagagagat agatttgtag agagagactg
     3301 gtgatttcag cgtaccgaca ctaaaccttc ctcttcttct tcggcccacc gtactcgtca
     3361 attccaaggg catcggtaaa catctgctca aactcgaagt cggccgtatc cagagcgccg
     3421 tagggggcgg agtcgtgggg ggtaaatccc ggccccgggg aatccccgtc ccccaacatg
     3481 tccagatcga aatcgtctag cgcgtcggca tgcgccatcg ccacgtcctc gccgtctaag
     3541 tggagctcgt cccccaggct gacgtcggtc gggggggcct tggcgtcggc ctgcttgatc
     3601 agttccagtg tcttttccac tctgtcggca ggcagaggtg ctcccagcac agccagggag
     3661 gcgtcagcca gcataatggc tgtctcgggc agagagtcgg cgctcagagg gctagggatc
     3721 ttgccgtcgt tccgccgcat ctcgatcacg ctcatggtga tgtggaaggg cagttcggct
     3781 ctggggtcgt cgcccacaat ctcggtggcc agatctctga acacgttggt cagggcctcg
     3841 cgctggctgt ggtactcggc gaattcctca gagcccacga tgggcagctg gtacagccgg
     3901 cccacgttcc acttggtgct cagcagcagc cgcacttcgc tggccacgat agcccacagt
     3961 ctcatttcgg gtccggcgtc cagggtagac agatcctcgg ccagcacggt ggagggttcc
     4021 acggtgcttt tcagcagggt caggaagatc tcggtcttgc tggggaagtg gtagtacagg
     4081 ctggcctgtc tgatgcccac agcgtcggca atctggtggg tgctggtggt ggcaaagccc
     4141 tgtctggtga acagctcggc gctggcatcc agaatctctt cccgggggtt cttgccggct
     4201 ctccgaggag cagatcttct gggtctgccc acggcgccag ccatggtggg gatcccagcg
     4261 tgtcctctcc aaatgaaatg aacttcctta tatagaggaa gggtcttgcg aaggatagtg
     4321 ggattgtgcg tcatccctta cgtcagtgga gatggatcct ctagaggcac gtggcggcac
     4381 gtggcggcac gtggcggcac gtggcaagct tggatccacg atatcctgca ggcatgcaag
     4441 cttagcttga gcttggatca gattgtcgtt tcccgccttc agtttaaact acattagaag
     4501 ccgagtcact acggttagtc cgtttgcatt ctgaatgttg agaacacagt aaaatggaat
     4561 gtgttttagt ttatattcaa atctcaatag aagaagagtc tattacaatg ataagtaata
     4621 gattagacac gaaaacgact aacttaagac agagaagaaa accaaaagcg atggagattt
     4681 ctttctttta ttgctgtgaa tgcataaagg aagtaaaaca caaagtgaac atcatctgcg
     4741 agagaatggc ttgacattaa gggtcctcga ccttacgctt tttcttaggt ttgtatagtt
     4801 catccatgcc atgtgtaatc ccagcagctg ttacaaactc aagaaggacc atgtgatctc
     4861 tcttttcgtt gggatctttc gaaagggcag attgtgtgga caggtaatgg ttgtctggta
     4921 aaaggacagg gccatcgcca attggagtat tttgttgata atgatcagcg agttgcacgc
     4981 cgccgtcctc gatgttgtgg cgggtcttga agttggcttt gatgccgttc ttttgcttgt
     5041 cggccatgat gtatacgttg tgggagttgt agttgtattc caacttgtgg ccgaggatgt
     5101 ttccgtcctc cttgaaatcg attcccttaa gctcgatcct gttgacgagg gtgtcaccct
     5161 caaacttgac ttcagcacgt gtcttgtagt tcccgtcgtc cttgaagaag atggtcctct
     5221 cctgcacgta tccctcaggc atggcgctct tgaagaagtc gtgccgcttc atatgatcag
     5281 ggtaacggga gaagcactgc acgccctgca catcaacaaa ttttggtcat atattagaaa
     5341 agttataaat taaaatatac acacttataa actacagaaa agcaattgct atatactaca
     5401 ttcttttatt ttgaaaaaaa tatttgaaat attatattac tactaattaa tgataattat
     5461 tatatatata tcaaaggtag aagcagaaac ttacgtaggt cagggtggtg accagggttg
     5521 gccatggaac aggtagtttt ccagtagtgc aaataaattt aagggtaagt tttccgtatg
     5581 ttgcatcacc ttcaccctct ccactgacag aaaatttgtg cccattaaca tcaccatcta
     5641 attcaacaag aattgggaca actccagtga aaagttcttc tcctttactc accatggtgt
     5701 ctagagtccc ccgtgttctc tccaaatgaa atgaacttcc ttatatagag gaagggtctt
     5761 gcgaaaggat agtgggattg tgggtggcct agctagctta gacccgatta tctatagatc
     5821 gatagaaatt cgcggccgca gccaactttt ctatcattag agaccctcga gggcgcgccc
     5881 cgatctagta acatagatga caccgcgcgc gataatttat cctagtttgc gcgctatatt
     5941 ttgttttcta tcgcgtatta aatgtataat tgcgggactc taatcataaa aacccatctc
     6001 ataaataacg tcatgcatta catgttaatt attacatgct taacgtaatt caacagaaat
     6061 tatatgataa tcatcgcaag accggcaaca ggattcaatc ttaagaaact ttattgccaa
     6121 atgtttgaac gatctgcagc cgggcggccg ctttacttgt acagctcctc catgccgccg
     6181 gtggagtggc ggccctcggc gcgttcgtac tgttccacga tggtgtagtc ctcgttgtgg
     6241 gaggtgatgt ccaacttgat gttgacgttg taggcgccgg gcagctgcac gggcttcttg
     6301 gccttgtagg tggtcttgac ctcagcgtcg tagtggccgc cgtccttcag cttcagcctc
     6361 tgcttgatct cgcccttcag ggcgccgtcc tcggggtaca tccgctcgga ggaggcctcc
     6421 cagcccatgg tcttattctg cattacgggg ccgtcggagg ggaagttggt gccgcgcagc
     6481 ttcaccttgt agatgaactc gccgtcctgc agggaggagt cctgggtcac ggtcaccacg
     6541 ccgccgtcct cgaagttcat cacgcgctcc cacttgaagc cctcggggaa ggacagcttc
     6601 aagtagtcgg ggatgtcggc ggggtgcttc acgtaggcct tggagccgta catgaactga
     6661 ggggacagga tgtcccaggc gaagggcagg gggccaccct tggtcacctt cagcttggcg
     6721 gtctgggtgc cctcgtaggg gcggccctcg ccctcgccct cgatctcgaa ctcgtggccg
     6781 ttcacggagc cctccatgtg caccttgaag cgcatgaact ccttgatgat ggccatgtta
     6841 tcctcctcgc ccttgctcac catggatccg acgttggcag cacttctgaa tgatccaaga
     6901 gacttagaac ctgaagctct ttgaacgaat tggtggtaga atgcagctat cgcagctcca
     6961 atgaatggtc caacccaaaa tatccagtgg tcatcccatg gcttgctctt gttgtagatt
     7021 acggcagctc cgaaactcct tgccgggtta attccggttc cggtaatggg aatggtagcc
     7081 aaatgtacca tgaacacggc aaatccgatt ggaagtggcg ccaacaccgg aacgtgggag
     7141 tctctggcac tacgtttggg gtcagtggca gagaatacgg tgtagacaag aacgaaagta
     7201 ccaatgatct ctgcggctag accggtccct gtgctgtagc catcggctag agagttggct
     7261 ccacctccgt aacgggtgta gtaagagctt tggaaggctt tgacaaaacc aactccacaa
     7321 atcgcaccca aacactgagc gattatgtac aatagggccc taggtaacga cactttacgt
     7381 gccaagaata gcccaaatgt caccgctggg ttaatgtgac caccagagat accggcggtg
     7441 cagtagacga ggatgaagat cataccacca aaggcccaag cgataccgag gattccaact
     7501 ccgccgcaat ctacgccacc ggcatcagta tcggactgaa tcttgtaacc gatgactgtc
     7561 aaaacggtga tgtataagaa gaggagagtg gctacgaact ctgcgataac tgctctgtag
     7621 aaagaccact tctttagctc cgctccatca ataaacggag ctggtggcgg atcttgatag
     7681 tctcttgtct gaaatccttc tccgggaacg gcttccacat cctttgccat ggtgctgtta
     7741 atcagaaaaa ctcagattaa tcgacaaatt cgatcgcaca aactagaaac taacaccaga
     7801 tctagataga aatcacaaat cgaagagtaa ttattcgaca aaactcaaat tatttgaaca
     7861 aatcggatga tatctatgaa accctaatcg agaattaaga tgatatctaa cgatcaaacc
     7921 cagaaaatcg tctacgatct aagattaaca gaatctaaac caaagaacat atacgaaatt
     7981 gggatcgaac gaaaacaaaa tcgaagattt tgagagaata aggaacacag aaatttacct
     8041 tgatcacggt agagagaatt gagagaaagt ttttaagatt ttgagaaatt gaaatctgaa
     8101 ttgtgaagaa gaagagctct ttgggtattg ttttatagaa gaagaagaag aaaagacgag
     8161 gacgactagg tcacgagaaa gctaaggcgg tgaagcaata gctaataata aaatgacacg
     8221 tgtattgagc gttgtttaca cgcaaagttg tttttggcta attgccttat ttttaggttg
     8281 aggaaaagta tttgtgcttt gagttgataa acacgactcg tgtgtgccgg ctgcaaccac
     8341 tttgacgccg tttattactg actcgtcgac aaccaaagct taacgttaca ccacaatata
     8401 tcctgccaag agtctaattc cggggatcgg aaatccag
//
