LOCUS       sequence-391175-        8923 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants 
            (proUBQ10::PIP2A-mCherry-NOSt_6XAmtRO-2XPhlFO::TetR-NLS-19St_proT::G
            FPint-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8923)
  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 8923)
  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: JAB3616
FEATURES             Location/Qualifiers
     source          1..8923
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    18..40
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     CDS             1563..2273
                     /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(1708..1726)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      2292..2544
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3466..3494)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          3634..3822
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     4278..4298
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    4604..4780
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(4836..4856)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(5532..5578)
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     primer_bind     complement(5905..5925)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5905..5923)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5944..5960)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5944..5960)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5957..5979)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5968..5984
                     /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(5992..5998,5999..6016,6017..6022))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6037..6058
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6175..6192)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6297..6321
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(6412..7000)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6492..6511)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7087..7881)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      8378..8813
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 ccccggaatt agactcttgg caggatatat tgtggtgtaa cgttaagctt tggttgtcga
       61 cgagtcagta ataaacggcg tcaaagtggt tgcagccggc acacacgagt cgtgtttatc
      121 aactcaaagc acaaatactt ttcctcaacc taaaaataag gcaattagcc aaaaacaact
      181 ttgcgtgtaa acaacgctca atacacgtgt cattttatta ttagctattg cttcaccgcc
      241 ttagctttct cgtgacctag tcgtcctcgt cttttcttct tcttcttcta taaaacaata
      301 cccaaagagc tcttcttctt cacaattcag atttcaattt ctcaaaatct taaaaacttt
      361 ctctcaattc tctctaccgt gatcaaggta aatttctgtg ttccttattc tctcaaaatc
      421 ttcgattttg ttttcgttcg atcccaattt cgtatatgtt ctttggttta gattctgtta
      481 atcttagatc gtagacgatt ttctgggttt gatcgttaga tatcatctta attctcgatt
      541 agggtttcat agatatcatc cgatttgttc aaataatttg agttttgtcg aataattact
      601 cttcgatttg tgatttctat ctagatctgg tgttagtttc tagtttgtgc gatcgaattt
      661 gtcgattaat ctgagttttt ctgattaaca gcaccatggc aaaggatgtg gaagccgttc
      721 ccggagaagg atttcagaca agagactatc aagatccgcc accagctccg tttattgatg
      781 gagcggagct aaagaagtgg tctttctaca gagcagttat cgcagagttc gtagccactc
      841 tcctcttctt atacatcacc gttttgacag tcatcggtta caagattcag tccgatactg
      901 atgccggtgg cgtagattgc ggcggagttg gaatcctcgg tatcgcttgg gcctttggtg
      961 gtatgatctt catcctcgtc tactgcaccg ccggtatctc tggtggtcac attaacccag
     1021 cggtgacatt tgggctattc ttggcacgta aagtgtcgtt acctagggcc ctattgtaca
     1081 taatcgctca gtgtttgggt gcgatttgtg gagttggttt tgtcaaagcc ttccaaagct
     1141 cttactacac ccgttacgga ggtggagcca actctctagc cgatggctac agcacaggga
     1201 ccggtctagc cgcagagatc attggtactt tcgttcttgt ctacaccgta ttctctgcca
     1261 ctgaccccaa acgtagtgcc agagactccc acgttccggt gttggcgcca cttccaatcg
     1321 gatttgccgt gttcatggta catttggcta ccattcccat taccggaacc ggaattaacc
     1381 cggcaaggag tttcggagct gccgtaatct acaacaagag caagccatgg gatgaccact
     1441 ggatattttg ggttggacca ttcattggag ctgcgatagc tgcattctac caccaattcg
     1501 ttctaagagc ttcaggttct aagtctcttg gatcattcag aagtgctgcc aacgtcggat
     1561 ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
     1621 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     1681 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
     1741 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
     1801 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
     1861 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
     1921 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     1981 taatgcagaa taagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
     2041 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     2101 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     2161 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     2221 aacgcgccga gggccgccac tccaccggcg gcatggagga gctgtacaag taaagcggcc
     2281 gcccggctgc agatcgttca aacatttggc aataaagttt cttaagattg aatcctgttg
     2341 ccggtcttgc gatgattatc atataatttc tgttgaatta cgttaagcat gtaataatta
     2401 acatgtaatg catgacgtta tttatgagat gggtttttat gattagagtc ccgcaattat
     2461 acatttaata cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg
     2521 cggtgtcatc tatgttacta gatcggggcg cgccctcgag ggtctctaat gggtccccag
     2581 atttgccttt tcaatttcag aaagaatgct aacccacaga tggttagaga ggcttacgca
     2641 gcagctctca tcaagacgat ctacccgagc aataatctcc aggaaatcaa ataccttccc
     2701 aagaaggtta aagatgcagt caaaagattc aggactaact gcatcaagaa cacagagaaa
     2761 gatatatttc tcaagatcag aagtactatt ccagtatgga cgattcaagg cttgcttcac
     2821 aaaccaaggc aagtaataga gattggagtc tctaaaaagg tagttcccac tgaatcaaag
     2881 gccatggagt caaagattca aatagaggac ctaacagaac tcgccgtaaa gactggcgaa
     2941 cagttcatac agagtctctt acgactcaat gacaagaaga aaatcttcgt caacatggtg
     3001 gagcacgaca cacttgtcta ctccaaaaat atcaaagata cagtctcaga agaccaaagg
     3061 gcaattgaga cttttcaaca aagggtaata tccggaaacc tcctcggatt ccattgccca
     3121 gctatctgtc actttattgt gaagatagtg gaaaaggaag gtggctccta caaatgccat
     3181 cattgcgata aaggaaaggc catcgttgaa gatgcctctg ccgacagtgg tcccaaagat
     3241 ggacccccac ccacgaggag catcgtggaa aaagaagacg ttccaaccac gtcttcaaag
     3301 caagtggatt gatgtgatat ctccactgac gtaagggatg acgcacaatc ccactatcct
     3361 tcgcaagata gataattcta tataaactct atcactgata gagtggagag aacacggggg
     3421 actctagaca ccatggtgag taaaggagaa gaacttttca ctggagttgt cccaattctt
     3481 gttgaattag atggtgatgt taatgggcac aaattttctg tcagtggaga gggtgaaggt
     3541 gatgcaacat acggaaaact tacccttaaa tttatttgca ctactggaaa actacctgtt
     3601 ccatggccaa ccctggtcac caccctgacc tacgtaagtt tctgcttcta cctttgatat
     3661 atatataata attatcatta attagtagta atataatatt tcaaatattt ttttcaaaat
     3721 aaaagaatgt agtatatagc aattgctttt ctgtagttta taagtgtgta tattttaatt
     3781 tataactttt ctaatatatg accaaaattt gttgatgtgc agggcgtgca gtgcttctcc
     3841 cgttaccctg atcatatgaa gcggcacgac ttcttcaaga gcgccatgcc tgagggatac
     3901 gtgcaggaga ggaccatctt cttcaaggac gacgggaact acaagacacg tgctgaagtc
     3961 aagtttgagg gtgacaccct cgtcaacagg atcgagctta agggaatcga tttcaaggag
     4021 gacggaaaca tcctcggcca caagttggaa tacaactaca actcccacaa cgtatacatc
     4081 atggccgaca agcaaaagaa cggcatcaaa gccaacttca agacccgcca caacatcgag
     4141 gacggcggcg tgcaactcgc tgatcattat caacaaaata ctccaattgg cgatggccct
     4201 gtccttttac cagacaacca ttacctgtcc acacaatctg ccctttcgaa agatcccaac
     4261 gaaaagagag atcacatggt ccttcttgag tttgtaacag ctgctgggat tacacatggc
     4321 atggatgaac tatacaaata atgtcaagcc attctctcgc agatgatgtt cactttgtgt
     4381 tttacttcct ttatgcattc acagcaataa aagaaagaaa tctccatcgc ttttggtttt
     4441 cttctctgtc ttaagttagt cgttttcgtg tctaatctat tacttatcat tgtaatagac
     4501 tcttcttcta ttgagatttg aatataaact aaaacacatt ccattttact gtgttctcaa
     4561 cattcagaat gcaaacggac taaccgtagt gactcggctt atcgatctgg attttagtac
     4621 tggattttgg ttttaggaat tagaaatttt attgatagaa gtattttaca aatacaaata
     4681 catactaagg gtttcttata tgctcaacac atgagcgaaa ccctataaga accctaattt
     4741 cccttatcgg gaaactactc acacattatt tatggagaaa atagagagag atagatttgt
     4801 agagagagac tggtgatttc agcgtaccga cactaaacct tcctcttctt cttcgggctg
     4861 ccggactcgc acttcagctg tttttcgagg ccacagatga tcagctcgag gccgaacaga
     4921 aaggcgggct cggcgccctg atgatcgaac agctcaatgg cctgtctcag cagagggggc
     4981 atgctgtcgg tggtgggtgt ctctctttcc tctttggcga cctggtgttc ctgatcttcc
     5041 agcacgcagc ccagtgtaaa gtggcccacg gcgctcaggg cgtacagggc attttccagg
     5101 ctgaagccct gctggcacag gaaggccagc tggttttcca gtgtctcgta ctgcttctcg
     5161 gtaggtctgg tgcccaggtg cactttggcg ccatcccggt gggacagcag agcgcatctg
     5221 aagctcttgg cgttgttccg caggaaatcc tgccagctct cgccttccag ggggcaaaag
     5281 tgggtgtggt gccggtccag catctcgatg gccagggcgt ccagcagggc tctcttgttc
     5341 ttcacgtgcc agtacagggt gggctgttcc acgcccagtt tctgagccag cttccgggtg
     5401 gtcaggccct cgatgcccac ttcgttgagc agttccaggg cgctgttgat cactttgctc
     5461 ttgtccagcc ggctcatggt gaccttaacg atacggtacg tttcgtatca attactctag
     5521 agtcccccgt gttctctcca aatgaaatga acttccttat atagaggaag ggtcttgcga
     5581 aaggatagtg ggattgtgac ctaccttaac gatacggtac gtttcgtatc atgctaggtg
     5641 gcctagctag cttagacccg attatctata gatcgataga aactattatc tatagatcga
     5701 tagaaaacat tatctataga tcgatagaaa tgattatcta tagatcgata gaaatgatta
     5761 tctatagatc gatagaaatc attatctata gatcgataga aattcgcggc cgcagccaac
     5821 ttttctatta ctcgcttcga gaccgggcta gagcggacta gttctagagc ggccgccacc
     5881 gcggtggagc tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
     5941 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     6001 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6061 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     6121 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     6181 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     6241 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatga aggccttgac
     6301 aggatatatt ggcgggtaaa ctaagtcgct gtatgtgttt gtttgagatc tcatgtgagc
     6361 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6421 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6481 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6541 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6601 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6661 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6721 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6781 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6841 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6901 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6961 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7021 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgcattc
     7081 taggtactaa aacaattcat ccagtaaaat ataatatttt attttctccc aatcaggctt
     7141 gatccccagt aagtcaaaaa atagctcgac atactgttct tccccgatat cctccctgat
     7201 cgaccggacg cagaaggcaa tgtcatacca cttgtccgcc ctgccgcttc tcccaagatc
     7261 aataaagcca cttactttgc catctttcac aaagatgttg ctgtcaccca ggtcgccgtg
     7321 ggaaaagaca agttcctctt cgggcttttc cgtctttaaa aaatcataca gctcgcgcgg
     7381 atctttaaat ggagtatctt cttcccagtt ttcgcaatcc acatcggcca gatcgttatt
     7441 cagtaagtaa tccaattcgg ctaagcggct gtctaagcta ttcgtatagg gacaatccga
     7501 tatgtcgatg gagtgaaaga gcctgatgca ctccgcatac agctcgataa tcttttcagg
     7561 gctttgttca tcttcatact cttccgagca aaggacgcca tcggcctcac tcatgagcag
     7621 attgctccag ccatcatgcc gttcaaagtg caggaccttt ggaacaggca gctttccttc
     7681 cagccatagc atcatgtcct tttcccgttc cacatcatag gtggtccctt tataccggct
     7741 gtccgtcatt tttaaatata ggttttcatt ttctcccacc agcttatata ccttagcagg
     7801 tgacattcct tccgtatctt ttacgcagcg gtatttttcg atcagttttt tcaattccgg
     7861 tgatattctc attttagcca tttattattt ccttcctctt ttctacagta tttaaagata
     7921 ccccaagaag ctaattataa caagacgaac tccaattcac tgttccttgc attctaaaac
     7981 cttaaatacc agaaaacagc tttttcaaag ttgttttcaa agttggcgta taacatagta
     8041 tcgacggagc cgattttgaa accgcggtga tcacaggcag caacgctctg tcatcgttac
     8101 aatcaacatg ctaccctccg cgagatcatc cgtgtttcaa acccggcagc ttattgtgca
     8161 atgtaacatc agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct
     8221 gagttgaagg atcagatcac gcatcttccc gacaacgcag accgttccgt ggcaaagcaa
     8281 aagttcaaaa tcaccaactg gtccacctac aacaaagctc tcatcaaccg tggctccctc
     8341 actttctggc tggatgatgg ggcgattcag gagatcccca tccaacagcc cgccgtcgag
     8401 cgggcttttt tatccccgga agcctgtgga tagagggtag ttatccacgt gaaaccgcta
     8461 atgccccgca aagccttgat tcacggggct ttccggcccg ctccaaaaac tatccacgtg
     8521 aaatcgctaa tcagggtacg tgaaatcgct aatcggagta cgtgaaatcg ctaataaggt
     8581 cacgtgaaat cgctaatcaa aaaggcacgt gagaacgcta atagcccttt cagatcaaca
     8641 gcttgcaaac acccctcgct ccggcaagta gttacagcaa gtagtatgtt caattagctt
     8701 ttcaattatg aatatatata tcaattattg gtcgcccttg gcttgtggac aatgcgctac
     8761 gcgcaccggc tccgcccgtg gacaaccgca agcggttgcc caccgtcgag cgcctttgcc
     8821 cacaacccgg cggccggccg caacagatcg ttttataaat tttttttttt gaaaaagaaa
     8881 aagcccgaaa ggcggcaacc tctcgggctt ctggatttcc gat
//
