LOCUS       sequence-393617-        8935 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants 
            (proUBQ10::PIP2A-mCherry-NOSt_6XAO-2XPO::TetR-NLS-19St_proT::GFPint-
            ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8935)
  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 8935)
  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: JAB3479
FEATURES             Location/Qualifiers
     source          1..8935
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    24..46
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     CDS             1569..2279
                     /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(1714..1732)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      2298..2550
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3472..3500)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          3640..3828
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     4284..4304
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     promoter        4863..4909
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             5585..5605
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5661..5837
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(5923..5943)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5923..5941)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5962..5978)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5962..5978)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5975..5997)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5986..6002
                     /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(6010..6016,6017..6034,6035..6040))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6055..6076
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6193..6210)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6315..6339
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(6430..7018)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6510..6529)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7105..7899)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      8396..8831
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 tccgatcccc ggaattagac tcttggcagg atatattgtg gtgtaacgtt aagctttggt
       61 tgtcgacgag tcagtaataa acggcgtcaa agtggttgca gccggcacac acgagtcgtg
      121 tttatcaact caaagcacaa atacttttcc tcaacctaaa aataaggcaa ttagccaaaa
      181 acaactttgc gtgtaaacaa cgctcaatac acgtgtcatt ttattattag ctattgcttc
      241 accgccttag ctttctcgtg acctagtcgt cctcgtcttt tcttcttctt cttctataaa
      301 acaataccca aagagctctt cttcttcaca attcagattt caatttctca aaatcttaaa
      361 aactttctct caattctctc taccgtgatc aaggtaaatt tctgtgttcc ttattctctc
      421 aaaatcttcg attttgtttt cgttcgatcc caatttcgta tatgttcttt ggtttagatt
      481 ctgttaatct tagatcgtag acgattttct gggtttgatc gttagatatc atcttaattc
      541 tcgattaggg tttcatagat atcatccgat ttgttcaaat aatttgagtt ttgtcgaata
      601 attactcttc gatttgtgat ttctatctag atctggtgtt agtttctagt ttgtgcgatc
      661 gaatttgtcg attaatctga gtttttctga ttaacagcac catggcaaag gatgtggaag
      721 ccgttcccgg agaaggattt cagacaagag actatcaaga tccgccacca gctccgttta
      781 ttgatggagc ggagctaaag aagtggtctt tctacagagc agttatcgca gagttcgtag
      841 ccactctcct cttcttatac atcaccgttt tgacagtcat cggttacaag attcagtccg
      901 atactgatgc cggtggcgta gattgcggcg gagttggaat cctcggtatc gcttgggcct
      961 ttggtggtat gatcttcatc ctcgtctact gcaccgccgg tatctctggt ggtcacatta
     1021 acccagcggt gacatttggg ctattcttgg cacgtaaagt gtcgttacct agggccctat
     1081 tgtacataat cgctcagtgt ttgggtgcga tttgtggagt tggttttgtc aaagccttcc
     1141 aaagctctta ctacacccgt tacggaggtg gagccaactc tctagccgat ggctacagca
     1201 cagggaccgg tctagccgca gagatcattg gtactttcgt tcttgtctac accgtattct
     1261 ctgccactga ccccaaacgt agtgccagag actcccacgt tccggtgttg gcgccacttc
     1321 caatcggatt tgccgtgttc atggtacatt tggctaccat tcccattacc ggaaccggaa
     1381 ttaacccggc aaggagtttc ggagctgccg taatctacaa caagagcaag ccatgggatg
     1441 accactggat attttgggtt ggaccattca ttggagctgc gatagctgca ttctaccacc
     1501 aattcgttct aagagcttca ggttctaagt ctcttggatc attcagaagt gctgccaacg
     1561 tcggatccat ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc
     1621 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
     1681 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
     1741 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg
     1801 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt
     1861 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
     1921 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
     1981 gccccgtaat gcagaataag accatgggct gggaggcctc ctccgagcgg atgtaccccg
     2041 aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact
     2101 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
     2161 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
     2221 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggaggagctg tacaagtaaa
     2281 gcggccgccc ggctgcagat cgttcaaaca tttggcaata aagtttctta agattgaatc
     2341 ctgttgccgg tcttgcgatg attatcatat aatttctgtt gaattacgtt aagcatgtaa
     2401 taattaacat gtaatgcatg acgttattta tgagatgggt ttttatgatt agagtcccgc
     2461 aattatacat ttaatacgcg atagaaaaca aaatatagcg cgcaaactag gataaattat
     2521 cgcgcgcggt gtcatctatg ttactagatc ggggcgcgcc ctcgagggtc tctaatgggt
     2581 ccccagattt gccttttcaa tttcagaaag aatgctaacc cacagatggt tagagaggct
     2641 tacgcagcag ctctcatcaa gacgatctac ccgagcaata atctccagga aatcaaatac
     2701 cttcccaaga aggttaaaga tgcagtcaaa agattcagga ctaactgcat caagaacaca
     2761 gagaaagata tatttctcaa gatcagaagt actattccag tatggacgat tcaaggcttg
     2821 cttcacaaac caaggcaagt aatagagatt ggagtctcta aaaaggtagt tcccactgaa
     2881 tcaaaggcca tggagtcaaa gattcaaata gaggacctaa cagaactcgc cgtaaagact
     2941 ggcgaacagt tcatacagag tctcttacga ctcaatgaca agaagaaaat cttcgtcaac
     3001 atggtggagc acgacacact tgtctactcc aaaaatatca aagatacagt ctcagaagac
     3061 caaagggcaa ttgagacttt tcaacaaagg gtaatatccg gaaacctcct cggattccat
     3121 tgcccagcta tctgtcactt tattgtgaag atagtggaaa aggaaggtgg ctcctacaaa
     3181 tgccatcatt gcgataaagg aaaggccatc gttgaagatg cctctgccga cagtggtccc
     3241 aaagatggac ccccacccac gaggagcatc gtggaaaaag aagacgttcc aaccacgtct
     3301 tcaaagcaag tggattgatg tgatatctcc actgacgtaa gggatgacgc acaatcccac
     3361 tatccttcgc aagatagata attctatata aactctatca ctgatagagt ggagagaaca
     3421 cgggggactc tagacaccat ggtgagtaaa ggagaagaac ttttcactgg agttgtccca
     3481 attcttgttg aattagatgg tgatgttaat gggcacaaat tttctgtcag tggagagggt
     3541 gaaggtgatg caacatacgg aaaacttacc cttaaattta tttgcactac tggaaaacta
     3601 cctgttccat ggccaaccct ggtcaccacc ctgacctacg taagtttctg cttctacctt
     3661 tgatatatat ataataatta tcattaatta gtagtaatat aatatttcaa atattttttt
     3721 caaaataaaa gaatgtagta tatagcaatt gcttttctgt agtttataag tgtgtatatt
     3781 ttaatttata acttttctaa tatatgacca aaatttgttg atgtgcaggg cgtgcagtgc
     3841 ttctcccgtt accctgatca tatgaagcgg cacgacttct tcaagagcgc catgcctgag
     3901 ggatacgtgc aggagaggac catcttcttc aaggacgacg ggaactacaa gacacgtgct
     3961 gaagtcaagt ttgagggtga caccctcgtc aacaggatcg agcttaaggg aatcgatttc
     4021 aaggaggacg gaaacatcct cggccacaag ttggaataca actacaactc ccacaacgta
     4081 tacatcatgg ccgacaagca aaagaacggc atcaaagcca acttcaagac ccgccacaac
     4141 atcgaggacg gcggcgtgca actcgctgat cattatcaac aaaatactcc aattggcgat
     4201 ggccctgtcc ttttaccaga caaccattac ctgtccacac aatctgccct ttcgaaagat
     4261 cccaacgaaa agagagatca catggtcctt cttgagtttg taacagctgc tgggattaca
     4321 catggcatgg atgaactata caaataatgt caagccattc tctcgcagat gatgttcact
     4381 ttgtgtttta cttcctttat gcattcacag caataaaaga aagaaatctc catcgctttt
     4441 ggttttcttc tctgtcttaa gttagtcgtt ttcgtgtcta atctattact tatcattgta
     4501 atagactctt cttctattga gatttgaata taaactaaaa cacattccat tttactgtgt
     4561 tctcaacatt cagaatgcaa acggactaac cgtagtgact cggcttctaa tgatagaaaa
     4621 gttggctgcg gccgcgaatt tctatcgatc tatagataat gatttctatc gatctataga
     4681 taatcatttc tatcgatcta tagataatca tttctatcga tctatagata atgttttcta
     4741 tcgatctata gataatagtt tctatcgatc tatagataat cgggtctaag ctagctaggc
     4801 cacctagcat gatacgaaac gtaccgtatc gttaaggtag gtcacaatcc cactatcctt
     4861 tcgcaagacc cttcctctat ataaggaagt tcatttcatt tggagagaac acgggggact
     4921 ctagagtaat tgatacgaaa cgtaccgtat cgttaaggtc accatgagcc ggctggacaa
     4981 gagcaaagtg atcaacagcg ccctggaact gctcaacgaa gtgggcatcg agggcctgac
     5041 cacccggaag ctggctcaga aactgggcgt ggaacagccc accctgtact ggcacgtgaa
     5101 gaacaagaga gccctgctgg acgccctggc catcgagatg ctggaccggc accacaccca
     5161 cttttgcccc ctggaaggcg agagctggca ggatttcctg cggaacaacg ccaagagctt
     5221 cagatgcgct ctgctgtccc accgggatgg cgccaaagtg cacctgggca ccagacctac
     5281 cgagaagcag tacgagacac tggaaaacca gctggccttc ctgtgccagc agggcttcag
     5341 cctggaaaat gccctgtacg ccctgagcgc cgtgggccac tttacactgg gctgcgtgct
     5401 ggaagatcag gaacaccagg tcgccaaaga ggaaagagag acacccacca ccgacagcat
     5461 gccccctctg ctgagacagg ccattgagct gttcgatcat cagggcgccg agcccgcctt
     5521 tctgttcggc ctcgagctga tcatctgtgg cctcgaaaaa cagctgaagt gcgagtccgg
     5581 cagcccgaag aagaagagga aggtttagtg tcggtacgct gaaatcacca gtctctctct
     5641 acaaatctat ctctctctat tttctccata aataatgtgt gagtagtttc ccgataaggg
     5701 aaattagggt tcttataggg tttcgctcat gtgttgagca tataagaaac ccttagtatg
     5761 tatttgtatt tgtaaaatac ttctatcaat aaaatttcta attcctaaaa ccaaaatcca
     5821 gtactaaaat ccagatcgat gcttgatact cgcttcgaga ccgggctaga gcggactagt
     5881 tctagagcgg ccgccaccgc ggtggagctc cagcttttgt tccctttagt gagggttaat
     5941 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6001 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6061 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6121 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6181 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6241 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6301 gaacatgaag gccttgacag gatatattgg cgggtaaact aagtcgctgt atgtgtttgt
     6361 ttgagatctc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6421 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6481 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6541 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6601 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6661 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6721 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6781 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6841 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     6901 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6961 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7021 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7081 gattttggtc atgcattcta ggtactaaaa caattcatcc agtaaaatat aatattttat
     7141 tttctcccaa tcaggcttga tccccagtaa gtcaaaaaat agctcgacat actgttcttc
     7201 cccgatatcc tccctgatcg accggacgca gaaggcaatg tcataccact tgtccgccct
     7261 gccgcttctc ccaagatcaa taaagccact tactttgcca tctttcacaa agatgttgct
     7321 gtcacccagg tcgccgtggg aaaagacaag ttcctcttcg ggcttttccg tctttaaaaa
     7381 atcatacagc tcgcgcggat ctttaaatgg agtatcttct tcccagtttt cgcaatccac
     7441 atcggccaga tcgttattca gtaagtaatc caattcggct aagcggctgt ctaagctatt
     7501 cgtataggga caatccgata tgtcgatgga gtgaaagagc ctgatgcact ccgcatacag
     7561 ctcgataatc ttttcagggc tttgttcatc ttcatactct tccgagcaaa ggacgccatc
     7621 ggcctcactc atgagcagat tgctccagcc atcatgccgt tcaaagtgca ggacctttgg
     7681 aacaggcagc tttccttcca gccatagcat catgtccttt tcccgttcca catcataggt
     7741 ggtcccttta taccggctgt ccgtcatttt taaatatagg ttttcatttt ctcccaccag
     7801 cttatatacc ttagcaggtg acattccttc cgtatctttt acgcagcggt atttttcgat
     7861 cagttttttc aattccggtg atattctcat tttagccatt tattatttcc ttcctctttt
     7921 ctacagtatt taaagatacc ccaagaagct aattataaca agacgaactc caattcactg
     7981 ttccttgcat tctaaaacct taaataccag aaaacagctt tttcaaagtt gttttcaaag
     8041 ttggcgtata acatagtatc gacggagccg attttgaaac cgcggtgatc acaggcagca
     8101 acgctctgtc atcgttacaa tcaacatgct accctccgcg agatcatccg tgtttcaaac
     8161 ccggcagctt attgtgcaat gtaacatcag agattttgag acacaacgtg gctttgttga
     8221 ataaatcgaa cttttgctga gttgaaggat cagatcacgc atcttcccga caacgcagac
     8281 cgttccgtgg caaagcaaaa gttcaaaatc accaactggt ccacctacaa caaagctctc
     8341 atcaaccgtg gctccctcac tttctggctg gatgatgggg cgattcagga gatccccatc
     8401 caacagcccg ccgtcgagcg ggctttttta tccccggaag cctgtggata gagggtagtt
     8461 atccacgtga aaccgctaat gccccgcaaa gccttgattc acggggcttt ccggcccgct
     8521 ccaaaaacta tccacgtgaa atcgctaatc agggtacgtg aaatcgctaa tcggagtacg
     8581 tgaaatcgct aataaggtca cgtgaaatcg ctaatcaaaa aggcacgtga gaacgctaat
     8641 agccctttca gatcaacagc ttgcaaacac ccctcgctcc ggcaagtagt tacagcaagt
     8701 agtatgttca attagctttt caattatgaa tatatatatc aattattggt cgcccttggc
     8761 ttgtggacaa tgcgctacgc gcaccggctc cgcccgtgga caaccgcaag cggttgccca
     8821 ccgtcgagcg cctttgccca caacccggcg gccggccgca acagatcgtt ttataaattt
     8881 ttttttttga aaaagaaaaa gcccgaaagg cggcaacctc tcgggcttct ggatt
//
