LOCUS       sequence-393426-        8534 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_5XAmtRO::GFPi
            nt-NLS-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8534)
  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 8534)
  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: JAB2603
FEATURES             Location/Qualifiers
     source          1..8534
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    17..39
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     CDS             1562..2272
                     /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(1707..1725)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      2291..2543
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        2759..2805
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     primer_bind     complement(2859..2887)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          3027..3215
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     3671..3691
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             3732..3752
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        4212..4258
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             4943..5176
                     /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             5177..5197
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5253..5429
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(5515..5535)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5515..5533)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5554..5570)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5554..5570)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5567..5589)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5578..5594
                     /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(5602..5608,5609..5626,5627..5632))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5647..5668
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5785..5802)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5907..5931
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(6022..6610)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6102..6121)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6697..7491)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      7988..8423
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 cccggaatta gactcttggc aggatatatt gtggtgtaac gttaagcttt ggttgtcgac
       61 gagtcagtaa taaacggcgt caaagtggtt gcagccggca cacacgagtc gtgtttatca
      121 actcaaagca caaatacttt tcctcaacct aaaaataagg caattagcca aaaacaactt
      181 tgcgtgtaaa caacgctcaa tacacgtgtc attttattat tagctattgc ttcaccgcct
      241 tagctttctc gtgacctagt cgtcctcgtc ttttcttctt cttcttctat aaaacaatac
      301 ccaaagagct cttcttcttc acaattcaga tttcaatttc tcaaaatctt aaaaactttc
      361 tctcaattct ctctaccgtg atcaaggtaa atttctgtgt tccttattct ctcaaaatct
      421 tcgattttgt tttcgttcga tcccaatttc gtatatgttc tttggtttag attctgttaa
      481 tcttagatcg tagacgattt tctgggtttg atcgttagat atcatcttaa ttctcgatta
      541 gggtttcata gatatcatcc gatttgttca aataatttga gttttgtcga ataattactc
      601 ttcgatttgt gatttctatc tagatctggt gttagtttct agtttgtgcg atcgaatttg
      661 tcgattaatc tgagtttttc tgattaacag caccatggca aaggatgtgg aagccgttcc
      721 cggagaagga tttcagacaa gagactatca agatccgcca ccagctccgt ttattgatgg
      781 agcggagcta aagaagtggt ctttctacag agcagttatc gcagagttcg tagccactct
      841 cctcttctta tacatcaccg ttttgacagt catcggttac aagattcagt ccgatactga
      901 tgccggtggc gtagattgcg gcggagttgg aatcctcggt atcgcttggg cctttggtgg
      961 tatgatcttc atcctcgtct actgcaccgc cggtatctct ggtggtcaca ttaacccagc
     1021 ggtgacattt gggctattct tggcacgtaa agtgtcgtta cctagggccc tattgtacat
     1081 aatcgctcag tgtttgggtg cgatttgtgg agttggtttt gtcaaagcct tccaaagctc
     1141 ttactacacc cgttacggag gtggagccaa ctctctagcc gatggctaca gcacagggac
     1201 cggtctagcc gcagagatca ttggtacttt cgttcttgtc tacaccgtat tctctgccac
     1261 tgaccccaaa cgtagtgcca gagactccca cgttccggtg ttggcgccac ttccaatcgg
     1321 atttgccgtg ttcatggtac atttggctac cattcccatt accggaaccg gaattaaccc
     1381 ggcaaggagt ttcggagctg ccgtaatcta caacaagagc aagccatggg atgaccactg
     1441 gatattttgg gttggaccat tcattggagc tgcgatagct gcattctacc accaattcgt
     1501 tctaagagct tcaggttcta agtctcttgg atcattcaga agtgctgcca acgtcggatc
     1561 catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
     1621 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     1681 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
     1741 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
     1801 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
     1861 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
     1921 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     1981 aatgcagaat aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     2041 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     2101 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     2161 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     2221 acgcgccgag ggccgccact ccaccggcgg catggaggag ctgtacaagt aaagcggccg
     2281 cccggctgca gatcgttcaa acatttggca ataaagtttc ttaagattga atcctgttgc
     2341 cggtcttgcg atgattatca tataatttct gttgaattac gttaagcatg taataattaa
     2401 catgtaatgc atgacgttat ttatgagatg ggtttttatg attagagtcc cgcaattata
     2461 catttaatac gcgatagaaa acaaaatata gcgcgcaaac taggataaat tatcgcgcgc
     2521 ggtgtcatct atgttactag atcggggcgc gccctcgagg gtctctaatg atagaaaagt
     2581 tggctgcggc cgcgaatttc tatcgatcta tagataatgt tttctatcga tctatagata
     2641 atgttttcta tcgatctata gataatagtt tctatcgatc tatagataat agtttctatc
     2701 gatctataga taatcgggtc taagctagct aggccaccca caatcccact atcctttcgc
     2761 aagacccttc ctctatataa ggaagttcat ttcatttgga gagaacacgg gggactctag
     2821 acaccatggt gagtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat
     2881 tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa
     2941 catacggaaa acttaccctt aaatttattt gcactactgg aaaactacct gttccatggc
     3001 caaccctggt caccaccctg acctacgtaa gtttctgctt ctacctttga tatatatata
     3061 ataattatca ttaattagta gtaatataat atttcaaata tttttttcaa aataaaagaa
     3121 tgtagtatat agcaattgct tttctgtagt ttataagtgt gtatatttta atttataact
     3181 tttctaatat atgaccaaaa tttgttgatg tgcagggcgt gcagtgcttc tcccgttacc
     3241 ctgatcatat gaagcggcac gacttcttca agagcgccat gcctgaggga tacgtgcagg
     3301 agaggaccat cttcttcaag gacgacggga actacaagac acgtgctgaa gtcaagtttg
     3361 agggtgacac cctcgtcaac aggatcgagc ttaagggaat cgatttcaag gaggacggaa
     3421 acatcctcgg ccacaagttg gaatacaact acaactccca caacgtatac atcatggccg
     3481 acaagcaaaa gaacggcatc aaagccaact tcaagacccg ccacaacatc gaggacggcg
     3541 gcgtgcaact cgctgatcat tatcaacaaa atactccaat tggcgatggc cctgtccttt
     3601 taccagacaa ccattacctg tccacacaat ctgccctttc gaaagatccc aacgaaaaga
     3661 gagatcacat ggtccttctt gagtttgtaa cagctgctgg gattacacat ggcatggatg
     3721 aactatacaa acctaagaaa aagcgtaagg tcgaggaccc ttaatgtcaa gccattctct
     3781 cgcagatgat gttcactttg tgttttactt cctttatgca ttcacagcaa taaaagaaag
     3841 aaatctccat cgcttttggt tttcttctct gtcttaagtt agtcgttttc gtgtctaatc
     3901 tattacttat cattgtaata gactcttctt ctattgagat ttgaatataa actaaaacac
     3961 attccatttt actgtgttct caacattcag aatgcaaacg gactaaccgt agtgactcgg
     4021 cttctaatgt agtttaaact gaaggcggga aacgacaatc tgatccaagc tcaagctaag
     4081 cttgcatgcc tgcaggatat cgtggatcca agcttgccac gtgccgccac gtgccgccac
     4141 gtgccgccac gtgcctctag aggatccatc tccactgacg taagggatga cgcacaatcc
     4201 cactatcctt cgcaagaccc ttcctctata taaggaagtt catttcattt ggagaggaca
     4261 cgctgggatc cccaccatgg ctggcgccgt gggcagaccc agaagatctg ctcctcggag
     4321 agccggcaag aacccccggg aagagattct ggatgccagc gccgagctgt tcaccagaca
     4381 gggctttgcc accaccagca cccaccagat tgccgacgct gtgggcatca gacaggccag
     4441 cctgtactac cacttcccca gcaagaccga gatcttcctg accctgctga aaagcaccgt
     4501 ggaaccctcc accgtgctgg ccgaggatct gtctaccctg gacgccggac ccgaaatgag
     4561 actgtgggct atcgtggcca gcgaagtgcg gctgctgctg agcaccaagt ggaacgtggg
     4621 ccggctgtac cagctgccca tcgtgggctc tgaggaattc gccgagtacc acagccagcg
     4681 cgaggccctg accaacgtgt tcagagatct ggccaccgag attgtgggcg acgaccccag
     4741 agccgaactg cccttccaca tcaccatgag cgtgatcgag atgcggcgga acgacggcaa
     4801 gatccctagc cctctgagcg ccgactctct gcccgagaca gccattatgc tggctgacgc
     4861 ctccctggct gtgctgggag cacctctgcc tgccgacaga gtggaaaaga cactggaact
     4921 gatcaagcag gccgacgcca aggccccccc gaccgacgtc agcctggggg acgagctcca
     4981 cttagacggc gaggacgtgg cgatggcgca tgccgacgcg ctagacgatt tcgatctgga
     5041 catgttgggg gacggggatt ccccggggcc gggatttacc ccccacgact ccgcccccta
     5101 cggcgctctg gatacggccg acttcgagtt tgagcagatg tttaccgatg cccttggaat
     5161 tgacgagtac ggtgggccga agaagaagag gaaggtttag tgtcggtacg ctgaaatcac
     5221 cagtctctct ctacaaatct atctctctct attttctcca taaataatgt gtgagtagtt
     5281 tcccgataag ggaaattagg gttcttatag ggtttcgctc atgtgttgag catataagaa
     5341 acccttagta tgtatttgta tttgtaaaat acttctatca ataaaatttc taattcctaa
     5401 aaccaaaatc cagtactaaa atccagatcg atgcttgata ctcgcttcga gaccgggcta
     5461 gagcggacta gttctagagc ggccgccacc gcggtggagc tccagctttt gttcccttta
     5521 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5581 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5641 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5701 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5761 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5821 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5881 taacgcagga aagaacatga aggccttgac aggatatatt ggcgggtaaa ctaagtcgct
     5941 gtatgtgttt gtttgagatc tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6001 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6061 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6121 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6181 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6241 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6301 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6361 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6421 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     6481 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6541 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6601 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6661 ctcacgttaa gggattttgg tcatgcattc taggtactaa aacaattcat ccagtaaaat
     6721 ataatatttt attttctccc aatcaggctt gatccccagt aagtcaaaaa atagctcgac
     6781 atactgttct tccccgatat cctccctgat cgaccggacg cagaaggcaa tgtcatacca
     6841 cttgtccgcc ctgccgcttc tcccaagatc aataaagcca cttactttgc catctttcac
     6901 aaagatgttg ctgtcaccca ggtcgccgtg ggaaaagaca agttcctctt cgggcttttc
     6961 cgtctttaaa aaatcataca gctcgcgcgg atctttaaat ggagtatctt cttcccagtt
     7021 ttcgcaatcc acatcggcca gatcgttatt cagtaagtaa tccaattcgg ctaagcggct
     7081 gtctaagcta ttcgtatagg gacaatccga tatgtcgatg gagtgaaaga gcctgatgca
     7141 ctccgcatac agctcgataa tcttttcagg gctttgttca tcttcatact cttccgagca
     7201 aaggacgcca tcggcctcac tcatgagcag attgctccag ccatcatgcc gttcaaagtg
     7261 caggaccttt ggaacaggca gctttccttc cagccatagc atcatgtcct tttcccgttc
     7321 cacatcatag gtggtccctt tataccggct gtccgtcatt tttaaatata ggttttcatt
     7381 ttctcccacc agcttatata ccttagcagg tgacattcct tccgtatctt ttacgcagcg
     7441 gtatttttcg atcagttttt tcaattccgg tgatattctc attttagcca tttattattt
     7501 ccttcctctt ttctacagta tttaaagata ccccaagaag ctaattataa caagacgaac
     7561 tccaattcac tgttccttgc attctaaaac cttaaatacc agaaaacagc tttttcaaag
     7621 ttgttttcaa agttggcgta taacatagta tcgacggagc cgattttgaa accgcggtga
     7681 tcacaggcag caacgctctg tcatcgttac aatcaacatg ctaccctccg cgagatcatc
     7741 cgtgtttcaa acccggcagc ttattgtgca atgtaacatc agagattttg agacacaacg
     7801 tggctttgtt gaataaatcg aacttttgct gagttgaagg atcagatcac gcatcttccc
     7861 gacaacgcag accgttccgt ggcaaagcaa aagttcaaaa tcaccaactg gtccacctac
     7921 aacaaagctc tcatcaaccg tggctccctc actttctggc tggatgatgg ggcgattcag
     7981 gagatcccca tccaacagcc cgccgtcgag cgggcttttt tatccccgga agcctgtgga
     8041 tagagggtag ttatccacgt gaaaccgcta atgccccgca aagccttgat tcacggggct
     8101 ttccggcccg ctccaaaaac tatccacgtg aaatcgctaa tcagggtacg tgaaatcgct
     8161 aatcggagta cgtgaaatcg ctaataaggt cacgtgaaat cgctaatcaa aaaggcacgt
     8221 gagaacgcta atagcccttt cagatcaaca gcttgcaaac acccctcgct ccggcaagta
     8281 gttacagcaa gtagtatgtt caattagctt ttcaattatg aatatatata tcaattattg
     8341 gtcgcccttg gcttgtggac aatgcgctac gcgcaccggc tccgcccgtg gacaaccgca
     8401 agcggttgcc caccgtcgag cgcctttgcc cacaacccgg cggccggccg caacagatcg
     8461 ttttataaat tttttttttt gaaaaagaaa aagcccgaaa ggcggcaacc tctcgggctt
     8521 ctggatttcc gatc
//
