LOCUS       sequence-389462-        7053 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants 
            (proUBQ10::PIP2A-mCherry-NOSt_6XRpaRO-min35S::GFPint-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7053)
  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 7053)
  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: JAB2511
FEATURES             Location/Qualifiers
     source          1..7053
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1539..2249
                     /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(1684..1702)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      2268..2520
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        2749..2795
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     primer_bind     complement(2849..2877)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          3017..3205
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     3661..3681
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4052..4072)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4052..4070)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4091..4107)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4091..4107)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4104..4126)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4115..4131
                     /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(4139..4145,4146..4163,4164..4169))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4184..4205
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4322..4339)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4444..4468
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4559..5147)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4639..4658)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5238..6026)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      6484..6919
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(7047..7053,1..16)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
ORIGIN
        1 atatattgtg gtgtaacgtt aagctttggt tgtcgacgag tcagtaataa acggcgtcaa
       61 agtggttgca gccggcacac acgagtcgtg tttatcaact caaagcacaa atacttttcc
      121 tcaacctaaa aataaggcaa ttagccaaaa acaactttgc gtgtaaacaa cgctcaatac
      181 acgtgtcatt ttattattag ctattgcttc accgccttag ctttctcgtg acctagtcgt
      241 cctcgtcttt tcttcttctt cttctataaa acaataccca aagagctctt cttcttcaca
      301 attcagattt caatttctca aaatcttaaa aactttctct caattctctc taccgtgatc
      361 aaggtaaatt tctgtgttcc ttattctctc aaaatcttcg attttgtttt cgttcgatcc
      421 caatttcgta tatgttcttt ggtttagatt ctgttaatct tagatcgtag acgattttct
      481 gggtttgatc gttagatatc atcttaattc tcgattaggg tttcatagat atcatccgat
      541 ttgttcaaat aatttgagtt ttgtcgaata attactcttc gatttgtgat ttctatctag
      601 atctggtgtt agtttctagt ttgtgcgatc gaatttgtcg attaatctga gtttttctga
      661 ttaacagcac catggcaaag gatgtggaag ccgttcccgg agaaggattt cagacaagag
      721 actatcaaga tccgccacca gctccgttta ttgatggagc ggagctaaag aagtggtctt
      781 tctacagagc agttatcgca gagttcgtag ccactctcct cttcttatac atcaccgttt
      841 tgacagtcat cggttacaag attcagtccg atactgatgc cggtggcgta gattgcggcg
      901 gagttggaat cctcggtatc gcttgggcct ttggtggtat gatcttcatc ctcgtctact
      961 gcaccgccgg tatctctggt ggtcacatta acccagcggt gacatttggg ctattcttgg
     1021 cacgtaaagt gtcgttacct agggccctat tgtacataat cgctcagtgt ttgggtgcga
     1081 tttgtggagt tggttttgtc aaagccttcc aaagctctta ctacacccgt tacggaggtg
     1141 gagccaactc tctagccgat ggctacagca cagggaccgg tctagccgca gagatcattg
     1201 gtactttcgt tcttgtctac accgtattct ctgccactga ccccaaacgt agtgccagag
     1261 actcccacgt tccggtgttg gcgccacttc caatcggatt tgccgtgttc atggtacatt
     1321 tggctaccat tcccattacc ggaaccggaa ttaacccggc aaggagtttc ggagctgccg
     1381 taatctacaa caagagcaag ccatgggatg accactggat attttgggtt ggaccattca
     1441 ttggagctgc gatagctgca ttctaccacc aattcgttct aagagcttca ggttctaagt
     1501 ctcttggatc attcagaagt gctgccaacg tcggatccat ggtgagcaag ggcgaggagg
     1561 ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga
     1621 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     1681 ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc
     1741 ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact
     1801 tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     1861 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga
     1921 agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaataag accatgggct
     1981 gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc
     2041 agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg
     2101 ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct
     2161 cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca
     2221 ccggcggcat ggaggagctg tacaagtaaa gcggccgccc ggctgcagat cgttcaaaca
     2281 tttggcaata aagtttctta agattgaatc ctgttgccgg tcttgcgatg attatcatat
     2341 aatttctgtt gaattacgtt aagcatgtaa taattaacat gtaatgcatg acgttattta
     2401 tgagatgggt ttttatgatt agagtcccgc aattatacat ttaatacgcg atagaaaaca
     2461 aaatatagcg cgcaaactag gataaattat cgcgcgcggt gtcatctatg ttactagatc
     2521 ggggcgcgcc ctcgaggaag acgtaatgat agaaaagttg gctgcggccg cgaaacctgt
     2581 ccgatcggac aatagaacct gtccgatcgg acaataggac ctgtccgatc ggacaataca
     2641 acctgtccga tcggacaata gtacctgtcc gatcggacaa taagacctgt ccgatcggac
     2701 aatacgggtc taagctagct aggccaccca caatcccact atcctttcgc aagacccttc
     2761 ctctatataa ggaagttcat ttcatttgga gagaacacgg gggactctag acaccatggt
     2821 gagtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga
     2881 tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa
     2941 acttaccctt aaatttattt gcactactgg aaaactacct gttccatggc caaccctggt
     3001 caccaccctg acctacgtaa gtttctgctt ctacctttga tatatatata ataattatca
     3061 ttaattagta gtaatataat atttcaaata tttttttcaa aataaaagaa tgtagtatat
     3121 agcaattgct tttctgtagt ttataagtgt gtatatttta atttataact tttctaatat
     3181 atgaccaaaa tttgttgatg tgcagggcgt gcagtgcttc tcccgttacc ctgatcatat
     3241 gaagcggcac gacttcttca agagcgccat gcctgaggga tacgtgcagg agaggaccat
     3301 cttcttcaag gacgacggga actacaagac acgtgctgaa gtcaagtttg agggtgacac
     3361 cctcgtcaac aggatcgagc ttaagggaat cgatttcaag gaggacggaa acatcctcgg
     3421 ccacaagttg gaatacaact acaactccca caacgtatac atcatggccg acaagcaaaa
     3481 gaacggcatc aaagccaact tcaagacccg ccacaacatc gaggacggcg gcgtgcaact
     3541 cgctgatcat tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa
     3601 ccattacctg tccacacaat ctgccctttc gaaagatccc aacgaaaaga gagatcacat
     3661 ggtccttctt gagtttgtaa cagctgctgg gattacacat ggcatggatg aactatacaa
     3721 ataatgtcaa gccattctct cgcagatgat gttcactttg tgttttactt cctttatgca
     3781 ttcacagcaa taaaagaaag aaatctccat cgcttttggt tttcttctct gtcttaagtt
     3841 agtcgttttc gtgtctaatc tattacttat cattgtaata gactcttctt ctattgagat
     3901 ttgaatataa actaaaacac attccatttt actgtgttct caacattcag aatgcaaacg
     3961 gactaaccgt agtgactcgg cttgagtctt cgggctagag cggactagtt ctagagcggc
     4021 cgccaccgcg gtggagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
     4081 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     4141 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     4201 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4261 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4321 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4381 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgaagg
     4441 ccttgacagg atatattggc gggtaaacta agtcgctgta tgtgtttgtt tgagatctca
     4501 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4561 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4621 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4681 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4741 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4801 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4861 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4921 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4981 actacggcta cactagaaga acagtatttg gtatctgtgc tctgctgaag ccagttacct
     5041 tcggaagaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5101 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5161 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5221 tgaattcttg ttagacatta tttgccgact accttggtga tctcgccttt cacgtagtga
     5281 acaaattctt ccaactgatc tgcgcgcgag gccaagcgat cttcttgtcc aagataagcc
     5341 tgcctagctt caagtatgac gggctgatac tgggccggca ggcgctccat tgcccagtcg
     5401 gcagcgacat ccttcggcgc gattttgccg gttactgcgc tgtaccaaat gcgggacaac
     5461 gtaagcacta catttcgctc atcgccagcc cagtcgggcg gcgagttcca tagcgttaag
     5521 gtttcattta gcgcctcaaa tagatcctgt tcaggaaccg gatcaaagag ttcctccgcc
     5581 gctggaccta ccaaggcaac gctatgttct cttgcttttg tcagcaagat agccagatca
     5641 atgtcgatcg tggctggctc gaagatacct gcaagaatgt cattgcgctg ccattctcca
     5701 aattgcagtt cgcgcttagc tggataacgc cacggaatga tgtcgtcgtg cacaacaatg
     5761 gtgacttcta cagcgcggag aatctcgctc tctccagggg aagccgaagt ttccaaaagg
     5821 tcgttgatca aagctcgccg cgttgtttca tcaagcctta cggtcaccgt aaccagcaaa
     5881 tcaatatcac tgtgtggctt caggccgcca tccactgcgg agccgtacaa atgtacggcc
     5941 agcaacgtcg gttcgagatg gcgctcgatg acgccaacta cctctgatag ttgagtcgat
     6001 acttcggcga tcaccgcttc cctcatgatg tttaactcct gaattaagcc gcgccgcgaa
     6061 gcggtgtcgg cttgaatgaa ttgttaggcg tcatcctgtg ctcccgagaa ccagtaccag
     6121 tacatcgctg tttcgttcga gacttgaggt ctagttttat acgtgaacag gtcaatgccg
     6181 ccgagagtaa agccacattt tgcgtacaaa ttgcaggcag gtacattgtt cgtttgtgcc
     6241 atgatgatat atttttatct tgtgcaatgt aacatcagag attttgagac acaacgtggc
     6301 tttgttgaat aaatcgaact tttgctgagt tgaaggatca gatcacgcat cttcccgaca
     6361 acgcagaccg ttccgtggca aagcaaaagt tcaaaatcac caactggtcc acctacaaca
     6421 aagctctcat caaccgtggc tccctcactt tctggctgga tgatggggcg attcaggaga
     6481 tccccatcca acagcccgcc gtcgagcggg cttttttatc cccggaagcc tgtggataga
     6541 gggtagttat ccacgtgaaa ccgctaatgc cccgcaaagc cttgattcac ggggctttcc
     6601 ggcccgctcc aaaaactatc cacgtgaaat cgctaatcag ggtacgtgaa atcgctaatc
     6661 ggagtacgtg aaatcgctaa taaggtcacg tgaaatcgct aatcaaaaag gcacgtgaga
     6721 acgctaatag ccctttcaga tcaacagctt gcaaacaccc ctcgctccgg caagtagtta
     6781 cagcaagtag tatgttcaat tagcttttca attatgaata tatatatcaa ttattggtcg
     6841 cccttggctt gtggacaatg cgctacgcgc accggctccg cccgtggaca accgcaagcg
     6901 gttgcccacc gtcgagcgcc tttgcccaca acccggcggc cggccgcaac agatcgtttt
     6961 ataaattttt ttttttgaaa aagaaaaagc ccgaaaggcg gcaacctctc gggcttctgg
     7021 atttccgatc cccggaatta gactcttggc agg
//
