LOCUS       sequence-393458-        8447 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-ERF2(AD)-19St_1XAmtRO(M
            2)::GFPint-NLS-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8447)
  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 8447)
  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: JAB3278
FEATURES             Location/Qualifiers
     source          1..8447
                     /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        2650..2696
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     primer_bind     complement(2750..2778)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          2918..3106
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     3562..3582
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             3623..3643
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        4103..4149
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             5077..5097
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5153..5329
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(5415..5435)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5415..5433)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5454..5470)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5454..5470)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5467..5489)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5478..5494
                     /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(5502..5508,5509..5526,5527..5532))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5547..5568
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5685..5702)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5807..5831
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5922..6510)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6002..6021)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6597..7391)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      7888..8323
                     /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 gaatttctgt cgatctctaa ataatcgggt ctaagctagc taggccaccc acaatcccac
     2641 tatcctttcg caagaccctt cctctatata aggaagttca tttcatttgg agagaacacg
     2701 ggggactcta gacaccatgg tgagtaaagg agaagaactt ttcactggag ttgtcccaat
     2761 tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga
     2821 aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc
     2881 tgttccatgg ccaaccctgg tcaccaccct gacctacgta agtttctgct tctacctttg
     2941 atatatatat aataattatc attaattagt agtaatataa tatttcaaat atttttttca
     3001 aaataaaaga atgtagtata tagcaattgc ttttctgtag tttataagtg tgtatatttt
     3061 aatttataac ttttctaata tatgaccaaa atttgttgat gtgcagggcg tgcagtgctt
     3121 ctcccgttac cctgatcata tgaagcggca cgacttcttc aagagcgcca tgcctgaggg
     3181 atacgtgcag gagaggacca tcttcttcaa ggacgacggg aactacaaga cacgtgctga
     3241 agtcaagttt gagggtgaca ccctcgtcaa caggatcgag cttaagggaa tcgatttcaa
     3301 ggaggacgga aacatcctcg gccacaagtt ggaatacaac tacaactccc acaacgtata
     3361 catcatggcc gacaagcaaa agaacggcat caaagccaac ttcaagaccc gccacaacat
     3421 cgaggacggc ggcgtgcaac tcgctgatca ttatcaacaa aatactccaa ttggcgatgg
     3481 ccctgtcctt ttaccagaca accattacct gtccacacaa tctgcccttt cgaaagatcc
     3541 caacgaaaag agagatcaca tggtccttct tgagtttgta acagctgctg ggattacaca
     3601 tggcatggat gaactataca aacctaagaa aaagcgtaag gtcgaggacc cttaatgtca
     3661 agccattctc tcgcagatga tgttcacttt gtgttttact tcctttatgc attcacagca
     3721 ataaaagaaa gaaatctcca tcgcttttgg ttttcttctc tgtcttaagt tagtcgtttt
     3781 cgtgtctaat ctattactta tcattgtaat agactcttct tctattgaga tttgaatata
     3841 aactaaaaca cattccattt tactgtgttc tcaacattca gaatgcaaac ggactaaccg
     3901 tagtgactcg gcttctaatg tagtttaaac tgaaggcggg aaacgacaat ctgatccaag
     3961 ctcaagctaa gcttgcatgc ctgcaggata tcgtggatcc aagcttgcca cgtgccgcca
     4021 cgtgccgcca cgtgccgcca cgtgcctcta gaggatccat ctccactgac gtaagggatg
     4081 acgcacaatc ccactatcct tcgcaagacc cttcctctat ataaggaagt tcatttcatt
     4141 tggagaggac acgctgggat ccccaccatg gctggcgccg tgggcagacc cagaagatct
     4201 gctcctcgga gagccggcaa gaacccccgg gaagagattc tggatgccag cgccgagctg
     4261 ttcaccagac agggctttgc caccaccagc acccaccaga ttgccgacgc tgtgggcatc
     4321 agacaggcca gcctgtacta ccacttcccc agcaagaccg agatcttcct gaccctgctg
     4381 aaaagcaccg tggaaccctc caccgtgctg gccgaggatc tgtctaccct ggacgccgga
     4441 cccgaaatga gactgtgggc tatcgtggcc agcgaagtgc ggctgctgct gagcaccaag
     4501 tggaacgtgg gccggctgta ccagctgccc atcgtgggct ctgaggaatt cgccgagtac
     4561 cacagccagc gcgaggccct gaccaacgtg ttcagagatc tggccaccga gattgtgggc
     4621 gacgacccca gagccgaact gcccttccac atcaccatga gcgtgatcga gatgcggcgg
     4681 aacgacggca agatccctag ccctctgagc gccgactctc tgcccgagac agccattatg
     4741 ctggctgacg cctccctggc tgtgctggga gcacctctgc ctgccgacag agtggaaaag
     4801 acactggaac tgatcaagca ggccgacgcc aagggaggag gtgaatccga ctacgctttg
     4861 ttggagtcga taacacgtca cttgctagga ggaggaggag agaacgagct gcgactcaat
     4921 gagtcaacac cgagttcgtg tttcacagag agttggggag gtttgccatt gaaagagaat
     4981 gattcagagg acatgttggt gtacggactc ctcaaagatg ccttccattt tgacacgtca
     5041 tcatcggact tgagctgtct ttttgatttt ccggcgccga agaagaagag gaaggtttag
     5101 tgtcggtacg ctgaaatcac cagtctctct ctacaaatct atctctctct attttctcca
     5161 taaataatgt gtgagtagtt tcccgataag ggaaattagg gttcttatag ggtttcgctc
     5221 atgtgttgag catataagaa acccttagta tgtatttgta tttgtaaaat acttctatca
     5281 ataaaatttc taattcctaa aaccaaaatc cagtactaaa atccagatcg atgcttgata
     5341 ctcgcttcga gaccgggcta gagcggacta gttctagagc ggccgccacc gcggtggagc
     5401 tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag
     5461 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     5521 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     5581 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     5641 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5701 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5761 ttatccacag aatcagggga taacgcagga aagaacatga aggccttgac aggatatatt
     5821 ggcgggtaaa ctaagtcgct gtatgtgttt gtttgagatc tcatgtgagc aaaaggccag
     5881 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5941 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6001 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6061 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     6121 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6181 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6241 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6301 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6361 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6421 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6481 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6541 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgcattc taggtactaa
     6601 aacaattcat ccagtaaaat ataatatttt attttctccc aatcaggctt gatccccagt
     6661 aagtcaaaaa atagctcgac atactgttct tccccgatat cctccctgat cgaccggacg
     6721 cagaaggcaa tgtcatacca cttgtccgcc ctgccgcttc tcccaagatc aataaagcca
     6781 cttactttgc catctttcac aaagatgttg ctgtcaccca ggtcgccgtg ggaaaagaca
     6841 agttcctctt cgggcttttc cgtctttaaa aaatcataca gctcgcgcgg atctttaaat
     6901 ggagtatctt cttcccagtt ttcgcaatcc acatcggcca gatcgttatt cagtaagtaa
     6961 tccaattcgg ctaagcggct gtctaagcta ttcgtatagg gacaatccga tatgtcgatg
     7021 gagtgaaaga gcctgatgca ctccgcatac agctcgataa tcttttcagg gctttgttca
     7081 tcttcatact cttccgagca aaggacgcca tcggcctcac tcatgagcag attgctccag
     7141 ccatcatgcc gttcaaagtg caggaccttt ggaacaggca gctttccttc cagccatagc
     7201 atcatgtcct tttcccgttc cacatcatag gtggtccctt tataccggct gtccgtcatt
     7261 tttaaatata ggttttcatt ttctcccacc agcttatata ccttagcagg tgacattcct
     7321 tccgtatctt ttacgcagcg gtatttttcg atcagttttt tcaattccgg tgatattctc
     7381 attttagcca tttattattt ccttcctctt ttctacagta tttaaagata ccccaagaag
     7441 ctaattataa caagacgaac tccaattcac tgttccttgc attctaaaac cttaaatacc
     7501 agaaaacagc tttttcaaag ttgttttcaa agttggcgta taacatagta tcgacggagc
     7561 cgattttgaa accgcggtga tcacaggcag caacgctctg tcatcgttac aatcaacatg
     7621 ctaccctccg cgagatcatc cgtgtttcaa acccggcagc ttattgtgca atgtaacatc
     7681 agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct gagttgaagg
     7741 atcagatcac gcatcttccc gacaacgcag accgttccgt ggcaaagcaa aagttcaaaa
     7801 tcaccaactg gtccacctac aacaaagctc tcatcaaccg tggctccctc actttctggc
     7861 tggatgatgg ggcgattcag gagatcccca tccaacagcc cgccgtcgag cgggcttttt
     7921 tatccccgga agcctgtgga tagagggtag ttatccacgt gaaaccgcta atgccccgca
     7981 aagccttgat tcacggggct ttccggcccg ctccaaaaac tatccacgtg aaatcgctaa
     8041 tcagggtacg tgaaatcgct aatcggagta cgtgaaatcg ctaataaggt cacgtgaaat
     8101 cgctaatcaa aaaggcacgt gagaacgcta atagcccttt cagatcaaca gcttgcaaac
     8161 acccctcgct ccggcaagta gttacagcaa gtagtatgtt caattagctt ttcaattatg
     8221 aatatatata tcaattattg gtcgcccttg gcttgtggac aatgcgctac gcgcaccggc
     8281 tccgcccgtg gacaaccgca agcggttgcc caccgtcgag cgcctttgcc cacaacccgg
     8341 cggccggccg caacagatcg ttttataaat tttttttttt gaaaaagaaa aagcccgaaa
     8401 ggcggcaacc tctcgggctt ctggatttcc gatccccgga attagac
//
