LOCUS       sequence-391162-        8486 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_3XAmtRO::GFPi
            nt-NLS-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8486)
  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 8486)
  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: JAB2601
FEATURES             Location/Qualifiers
     source          1..8486
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    4..26
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     CDS             1549..2259
                     /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(1694..1712)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      2278..2530
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        2698..2744
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     primer_bind     complement(2798..2826)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          2966..3154
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     3610..3630
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             3671..3691
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        4151..4197
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             4882..5115
                     /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             5116..5136
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5192..5368
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(5454..5474)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5454..5472)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5493..5509)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5493..5509)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5506..5528)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5517..5533
                     /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(5541..5547,5548..5565,5566..5571))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5586..5607
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5724..5741)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5846..5870
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5961..6549)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6041..6060)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6636..7430)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      7927..8362
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 tcttggcagg atatattgtg gtgtaacgtt aagctttggt tgtcgacgag tcagtaataa
       61 acggcgtcaa agtggttgca gccggcacac acgagtcgtg tttatcaact caaagcacaa
      121 atacttttcc tcaacctaaa aataaggcaa ttagccaaaa acaactttgc gtgtaaacaa
      181 cgctcaatac acgtgtcatt ttattattag ctattgcttc accgccttag ctttctcgtg
      241 acctagtcgt cctcgtcttt tcttcttctt cttctataaa acaataccca aagagctctt
      301 cttcttcaca attcagattt caatttctca aaatcttaaa aactttctct caattctctc
      361 taccgtgatc aaggtaaatt tctgtgttcc ttattctctc aaaatcttcg attttgtttt
      421 cgttcgatcc caatttcgta tatgttcttt ggtttagatt ctgttaatct tagatcgtag
      481 acgattttct gggtttgatc gttagatatc atcttaattc tcgattaggg tttcatagat
      541 atcatccgat ttgttcaaat aatttgagtt ttgtcgaata attactcttc gatttgtgat
      601 ttctatctag atctggtgtt agtttctagt ttgtgcgatc gaatttgtcg attaatctga
      661 gtttttctga ttaacagcac catggcaaag gatgtggaag ccgttcccgg agaaggattt
      721 cagacaagag actatcaaga tccgccacca gctccgttta ttgatggagc ggagctaaag
      781 aagtggtctt tctacagagc agttatcgca gagttcgtag ccactctcct cttcttatac
      841 atcaccgttt tgacagtcat cggttacaag attcagtccg atactgatgc cggtggcgta
      901 gattgcggcg gagttggaat cctcggtatc gcttgggcct ttggtggtat gatcttcatc
      961 ctcgtctact gcaccgccgg tatctctggt ggtcacatta acccagcggt gacatttggg
     1021 ctattcttgg cacgtaaagt gtcgttacct agggccctat tgtacataat cgctcagtgt
     1081 ttgggtgcga tttgtggagt tggttttgtc aaagccttcc aaagctctta ctacacccgt
     1141 tacggaggtg gagccaactc tctagccgat ggctacagca cagggaccgg tctagccgca
     1201 gagatcattg gtactttcgt tcttgtctac accgtattct ctgccactga ccccaaacgt
     1261 agtgccagag actcccacgt tccggtgttg gcgccacttc caatcggatt tgccgtgttc
     1321 atggtacatt tggctaccat tcccattacc ggaaccggaa ttaacccggc aaggagtttc
     1381 ggagctgccg taatctacaa caagagcaag ccatgggatg accactggat attttgggtt
     1441 ggaccattca ttggagctgc gatagctgca ttctaccacc aattcgttct aagagcttca
     1501 ggttctaagt ctcttggatc attcagaagt gctgccaacg tcggatccat ggtgagcaag
     1561 ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag
     1621 ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
     1681 ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac
     1741 atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc
     1801 cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
     1861 gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc
     1921 tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaataag
     1981 accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc
     2041 gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc
     2101 acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg
     2161 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc
     2221 cgccactcca ccggcggcat ggaggagctg tacaagtaaa gcggccgccc ggctgcagat
     2281 cgttcaaaca tttggcaata aagtttctta agattgaatc ctgttgccgg tcttgcgatg
     2341 attatcatat aatttctgtt gaattacgtt aagcatgtaa taattaacat gtaatgcatg
     2401 acgttattta tgagatgggt ttttatgatt agagtcccgc aattatacat ttaatacgcg
     2461 atagaaaaca aaatatagcg cgcaaactag gataaattat cgcgcgcggt gtcatctatg
     2521 ttactagatc ggggcgcgcc ctcgagggtc tctaatgata gaaaagttgg ctgcggccgc
     2581 gaatttctat cgatctatag ataatgtttt ctatcgatct atagataata gtttctatcg
     2641 atctatagat aatcgggtct aagctagcta ggccacccac aatcccacta tcctttcgca
     2701 agacccttcc tctatataag gaagttcatt tcatttggag agaacacggg ggactctaga
     2761 caccatggtg agtaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt
     2821 agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgcaac
     2881 atacggaaaa cttaccctta aatttatttg cactactgga aaactacctg ttccatggcc
     2941 aaccctggtc accaccctga cctacgtaag tttctgcttc tacctttgat atatatataa
     3001 taattatcat taattagtag taatataata tttcaaatat ttttttcaaa ataaaagaat
     3061 gtagtatata gcaattgctt ttctgtagtt tataagtgtg tatattttaa tttataactt
     3121 ttctaatata tgaccaaaat ttgttgatgt gcagggcgtg cagtgcttct cccgttaccc
     3181 tgatcatatg aagcggcacg acttcttcaa gagcgccatg cctgagggat acgtgcagga
     3241 gaggaccatc ttcttcaagg acgacgggaa ctacaagaca cgtgctgaag tcaagtttga
     3301 gggtgacacc ctcgtcaaca ggatcgagct taagggaatc gatttcaagg aggacggaaa
     3361 catcctcggc cacaagttgg aatacaacta caactcccac aacgtataca tcatggccga
     3421 caagcaaaag aacggcatca aagccaactt caagacccgc cacaacatcg aggacggcgg
     3481 cgtgcaactc gctgatcatt atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt
     3541 accagacaac cattacctgt ccacacaatc tgccctttcg aaagatccca acgaaaagag
     3601 agatcacatg gtccttcttg agtttgtaac agctgctggg attacacatg gcatggatga
     3661 actatacaaa cctaagaaaa agcgtaaggt cgaggaccct taatgtcaag ccattctctc
     3721 gcagatgatg ttcactttgt gttttacttc ctttatgcat tcacagcaat aaaagaaaga
     3781 aatctccatc gcttttggtt ttcttctctg tcttaagtta gtcgttttcg tgtctaatct
     3841 attacttatc attgtaatag actcttcttc tattgagatt tgaatataaa ctaaaacaca
     3901 ttccatttta ctgtgttctc aacattcaga atgcaaacgg actaaccgta gtgactcggc
     3961 ttctaatgta gtttaaactg aaggcgggaa acgacaatct gatccaagct caagctaagc
     4021 ttgcatgcct gcaggatatc gtggatccaa gcttgccacg tgccgccacg tgccgccacg
     4081 tgccgccacg tgcctctaga ggatccatct ccactgacgt aagggatgac gcacaatccc
     4141 actatccttc gcaagaccct tcctctatat aaggaagttc atttcatttg gagaggacac
     4201 gctgggatcc ccaccatggc tggcgccgtg ggcagaccca gaagatctgc tcctcggaga
     4261 gccggcaaga acccccggga agagattctg gatgccagcg ccgagctgtt caccagacag
     4321 ggctttgcca ccaccagcac ccaccagatt gccgacgctg tgggcatcag acaggccagc
     4381 ctgtactacc acttccccag caagaccgag atcttcctga ccctgctgaa aagcaccgtg
     4441 gaaccctcca ccgtgctggc cgaggatctg tctaccctgg acgccggacc cgaaatgaga
     4501 ctgtgggcta tcgtggccag cgaagtgcgg ctgctgctga gcaccaagtg gaacgtgggc
     4561 cggctgtacc agctgcccat cgtgggctct gaggaattcg ccgagtacca cagccagcgc
     4621 gaggccctga ccaacgtgtt cagagatctg gccaccgaga ttgtgggcga cgaccccaga
     4681 gccgaactgc ccttccacat caccatgagc gtgatcgaga tgcggcggaa cgacggcaag
     4741 atccctagcc ctctgagcgc cgactctctg cccgagacag ccattatgct ggctgacgcc
     4801 tccctggctg tgctgggagc acctctgcct gccgacagag tggaaaagac actggaactg
     4861 atcaagcagg ccgacgccaa ggcccccccg accgacgtca gcctggggga cgagctccac
     4921 ttagacggcg aggacgtggc gatggcgcat gccgacgcgc tagacgattt cgatctggac
     4981 atgttggggg acggggattc cccggggccg ggatttaccc cccacgactc cgccccctac
     5041 ggcgctctgg atacggccga cttcgagttt gagcagatgt ttaccgatgc ccttggaatt
     5101 gacgagtacg gtgggccgaa gaagaagagg aaggtttagt gtcggtacgc tgaaatcacc
     5161 agtctctctc tacaaatcta tctctctcta ttttctccat aaataatgtg tgagtagttt
     5221 cccgataagg gaaattaggg ttcttatagg gtttcgctca tgtgttgagc atataagaaa
     5281 cccttagtat gtatttgtat ttgtaaaata cttctatcaa taaaatttct aattcctaaa
     5341 accaaaatcc agtactaaaa tccagatcga tgcttgatac tcgcttcgag accgggctag
     5401 agcggactag ttctagagcg gccgccaccg cggtggagct ccagcttttg ttccctttag
     5461 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     5521 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     5581 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5641 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5701 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5761 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5821 aacgcaggaa agaacatgaa ggccttgaca ggatatattg gcgggtaaac taagtcgctg
     5881 tatgtgtttg tttgagatct catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5941 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6001 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6061 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6121 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     6181 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6241 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6301 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6361 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     6421 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6481 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6541 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6601 tcacgttaag ggattttggt catgcattct aggtactaaa acaattcatc cagtaaaata
     6661 taatatttta ttttctccca atcaggcttg atccccagta agtcaaaaaa tagctcgaca
     6721 tactgttctt ccccgatatc ctccctgatc gaccggacgc agaaggcaat gtcataccac
     6781 ttgtccgccc tgccgcttct cccaagatca ataaagccac ttactttgcc atctttcaca
     6841 aagatgttgc tgtcacccag gtcgccgtgg gaaaagacaa gttcctcttc gggcttttcc
     6901 gtctttaaaa aatcatacag ctcgcgcgga tctttaaatg gagtatcttc ttcccagttt
     6961 tcgcaatcca catcggccag atcgttattc agtaagtaat ccaattcggc taagcggctg
     7021 tctaagctat tcgtataggg acaatccgat atgtcgatgg agtgaaagag cctgatgcac
     7081 tccgcataca gctcgataat cttttcaggg ctttgttcat cttcatactc ttccgagcaa
     7141 aggacgccat cggcctcact catgagcaga ttgctccagc catcatgccg ttcaaagtgc
     7201 aggacctttg gaacaggcag ctttccttcc agccatagca tcatgtcctt ttcccgttcc
     7261 acatcatagg tggtcccttt ataccggctg tccgtcattt ttaaatatag gttttcattt
     7321 tctcccacca gcttatatac cttagcaggt gacattcctt ccgtatcttt tacgcagcgg
     7381 tatttttcga tcagtttttt caattccggt gatattctca ttttagccat ttattatttc
     7441 cttcctcttt tctacagtat ttaaagatac cccaagaagc taattataac aagacgaact
     7501 ccaattcact gttccttgca ttctaaaacc ttaaatacca gaaaacagct ttttcaaagt
     7561 tgttttcaaa gttggcgtat aacatagtat cgacggagcc gattttgaaa ccgcggtgat
     7621 cacaggcagc aacgctctgt catcgttaca atcaacatgc taccctccgc gagatcatcc
     7681 gtgtttcaaa cccggcagct tattgtgcaa tgtaacatca gagattttga gacacaacgt
     7741 ggctttgttg aataaatcga acttttgctg agttgaagga tcagatcacg catcttcccg
     7801 acaacgcaga ccgttccgtg gcaaagcaaa agttcaaaat caccaactgg tccacctaca
     7861 acaaagctct catcaaccgt ggctccctca ctttctggct ggatgatggg gcgattcagg
     7921 agatccccat ccaacagccc gccgtcgagc gggctttttt atccccggaa gcctgtggat
     7981 agagggtagt tatccacgtg aaaccgctaa tgccccgcaa agccttgatt cacggggctt
     8041 tccggcccgc tccaaaaact atccacgtga aatcgctaat cagggtacgt gaaatcgcta
     8101 atcggagtac gtgaaatcgc taataaggtc acgtgaaatc gctaatcaaa aaggcacgtg
     8161 agaacgctaa tagccctttc agatcaacag cttgcaaaca cccctcgctc cggcaagtag
     8221 ttacagcaag tagtatgttc aattagcttt tcaattatga atatatatat caattattgg
     8281 tcgcccttgg cttgtggaca atgcgctacg cgcaccggct ccgcccgtgg acaaccgcaa
     8341 gcggttgccc accgtcgagc gcctttgccc acaacccggc ggccggccgc aacagatcgt
     8401 tttataaatt tttttttttg aaaaagaaaa agcccgaaag gcggcaacct ctcgggcttc
     8461 tggatttccg atccccggaa ttagac
//
