LOCUS       sequence-389434-        7053 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::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: JAB2144
FEATURES             Location/Qualifiers
     source          1..7053
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..23
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     CDS             1546..2256
                     /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(1691..1709)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      2275..2527
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        2806..2852
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     primer_bind     complement(2940..2968)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3668..3688
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4059..4079)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4059..4077)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4098..4114)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4098..4114)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4111..4133)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4122..4138
                     /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(4146..4152,4153..4170,4171..4176))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4191..4212
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4329..4346)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4451..4475
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4566..5154)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4646..4665)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5245..6033)
                     /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      6491..6926
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 tggcaggata tattgtggtg taacgttaag ctttggttgt cgacgagtca gtaataaacg
       61 gcgtcaaagt ggttgcagcc ggcacacacg agtcgtgttt atcaactcaa agcacaaata
      121 cttttcctca acctaaaaat aaggcaatta gccaaaaaca actttgcgtg taaacaacgc
      181 tcaatacacg tgtcatttta ttattagcta ttgcttcacc gccttagctt tctcgtgacc
      241 tagtcgtcct cgtcttttct tcttcttctt ctataaaaca atacccaaag agctcttctt
      301 cttcacaatt cagatttcaa tttctcaaaa tcttaaaaac tttctctcaa ttctctctac
      361 cgtgatcaag gtaaatttct gtgttcctta ttctctcaaa atcttcgatt ttgttttcgt
      421 tcgatcccaa tttcgtatat gttctttggt ttagattctg ttaatcttag atcgtagacg
      481 attttctggg tttgatcgtt agatatcatc ttaattctcg attagggttt catagatatc
      541 atccgatttg ttcaaataat ttgagttttg tcgaataatt actcttcgat ttgtgatttc
      601 tatctagatc tggtgttagt ttctagtttg tgcgatcgaa tttgtcgatt aatctgagtt
      661 tttctgatta acagcaccat ggcaaaggat gtggaagccg ttcccggaga aggatttcag
      721 acaagagact atcaagatcc gccaccagct ccgtttattg atggagcgga gctaaagaag
      781 tggtctttct acagagcagt tatcgcagag ttcgtagcca ctctcctctt cttatacatc
      841 accgttttga cagtcatcgg ttacaagatt cagtccgata ctgatgccgg tggcgtagat
      901 tgcggcggag ttggaatcct cggtatcgct tgggcctttg gtggtatgat cttcatcctc
      961 gtctactgca ccgccggtat ctctggtggt cacattaacc cagcggtgac atttgggcta
     1021 ttcttggcac gtaaagtgtc gttacctagg gccctattgt acataatcgc tcagtgtttg
     1081 ggtgcgattt gtggagttgg ttttgtcaaa gccttccaaa gctcttacta cacccgttac
     1141 ggaggtggag ccaactctct agccgatggc tacagcacag ggaccggtct agccgcagag
     1201 atcattggta ctttcgttct tgtctacacc gtattctctg ccactgaccc caaacgtagt
     1261 gccagagact cccacgttcc ggtgttggcg ccacttccaa tcggatttgc cgtgttcatg
     1321 gtacatttgg ctaccattcc cattaccgga accggaatta acccggcaag gagtttcgga
     1381 gctgccgtaa tctacaacaa gagcaagcca tgggatgacc actggatatt ttgggttgga
     1441 ccattcattg gagctgcgat agctgcattc taccaccaat tcgttctaag agcttcaggt
     1501 tctaagtctc ttggatcatt cagaagtgct gccaacgtcg gatccatggt gagcaagggc
     1561 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
     1621 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     1681 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
     1741 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
     1801 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     1861 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
     1921 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaataagacc
     1981 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
     2041 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
     2101 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac
     2161 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
     2221 cactccaccg gcggcatgga ggagctgtac aagtaaagcg gccgcccggc tgcagatcgt
     2281 tcaaacattt ggcaataaag tttcttaaga ttgaatcctg ttgccggtct tgcgatgatt
     2341 atcatataat ttctgttgaa ttacgttaag catgtaataa ttaacatgta atgcatgacg
     2401 ttatttatga gatgggtttt tatgattaga gtcccgcaat tatacattta atacgcgata
     2461 gaaaacaaaa tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc atctatgtta
     2521 ctagatcggg gcgcgccctc gaggaagacg taatgataga aaagttggct gcggccgcga
     2581 atttctatcg atctatagat aatgatttct atcgatctat agataatcat ttctatcgat
     2641 ctatagataa tcatttctat cgatctatag ataatgtttt ctatcgatct atagataata
     2701 gtttctatcg atctatagat aatcgggtct aagctagcta ggccacctag catgatacga
     2761 aacgtaccgt atcgttaagg taggtcacaa tcccactatc ctttcgcaag acccttcctc
     2821 tatataagga agttcatttc atttggagag aacacggggg actctagagt aattgatacg
     2881 aaacgtaccg tatcgttaag gtcaccatgg tgagtaaagg agaagaactt ttcactggag
     2941 ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg
     3001 gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg
     3061 gaaaactacc tgttccatgg ccaaccctgg tcaccaccct gacctacgta agtttctgct
     3121 tctacctttg atatatatat aataagttta taagtgtgta tattttaatt tataactttt
     3181 ctaatatatg accaaaattt gttgatgtgc agggcgtgca gtgcttctcc cgttaccctg
     3241 atcatatgaa gcggcacgac ttcttcaaga gcgccatgcc tgagggatac gtgcaggaga
     3301 ggaccatctt cttcaaggac gacgggaact acaagacacg tgctgaagtc aagtttgagg
     3361 gtgacaccct cgtcaacagg atcgagctta agggaatcga tttcaaggag gacggaaaca
     3421 tcctcggcca caagttggaa tacaactaca actcccacaa cgtatacatc atggccgaca
     3481 agcaaaagaa cggcatcaaa gccaacttca agacccgcca caacatcgag gacggcggcg
     3541 tgcaactcgc tgatcattat caacaaaata ctccaattgg cgatggccct gtccttttac
     3601 cagacaacca ttacctgtcc acacaatctg ccctttcgaa agatcccaac gaaaagagag
     3661 atcacatggt ccttcttgag tttgtaacag ctgctgggat tacacatggc atggatgaac
     3721 tatacaaata atgtcaagcc attctctcgc agatgatgtt cactttgtgt tttacttcct
     3781 ttatgcattc acagcaataa aagaaagaaa tctccatcgc ttttggtttt cttctctgtc
     3841 ttaagttagt cgttttcgtg tctaatctat tacttatcat tgtaatagac tcttcttcta
     3901 ttgagatttg aatataaact aaaacacatt ccattttact gtgttctcaa cattcagaat
     3961 gcaaacggac taaccgtagt gactcggctt gagtcttcgg gctagagcgg actagttcta
     4021 gagcggccgc caccgcggtg gagctccagc ttttgttccc tttagtgagg gttaattgcg
     4081 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     4141 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4201 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4261 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4321 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4381 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4441 atgaaggcct tgacaggata tattggcggg taaactaagt cgctgtatgt gtttgtttga
     4501 gatctcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4561 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4621 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4681 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4741 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4801 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4861 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4921 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4981 tggcctaact acggctacac tagaagaaca gtatttggta tctgtgctct gctgaagcca
     5041 gttaccttcg gaagaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5101 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5161 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5221 ttggtcatga attcttgtta gacattattt gccgactacc ttggtgatct cgcctttcac
     5281 gtagtgaaca aattcttcca actgatctgc gcgcgaggcc aagcgatctt cttgtccaag
     5341 ataagcctgc ctagcttcaa gtatgacggg ctgatactgg gccggcaggc gctccattgc
     5401 ccagtcggca gcgacatcct tcggcgcgat tttgccggtt actgcgctgt accaaatgcg
     5461 ggacaacgta agcactacat ttcgctcatc gccagcccag tcgggcggcg agttccatag
     5521 cgttaaggtt tcatttagcg cctcaaatag atcctgttca ggaaccggat caaagagttc
     5581 ctccgccgct ggacctacca aggcaacgct atgttctctt gcttttgtca gcaagatagc
     5641 cagatcaatg tcgatcgtgg ctggctcgaa gatacctgca agaatgtcat tgcgctgcca
     5701 ttctccaaat tgcagttcgc gcttagctgg ataacgccac ggaatgatgt cgtcgtgcac
     5761 aacaatggtg acttctacag cgcggagaat ctcgctctct ccaggggaag ccgaagtttc
     5821 caaaaggtcg ttgatcaaag ctcgccgcgt tgtttcatca agccttacgg tcaccgtaac
     5881 cagcaaatca atatcactgt gtggcttcag gccgccatcc actgcggagc cgtacaaatg
     5941 tacggccagc aacgtcggtt cgagatggcg ctcgatgacg ccaactacct ctgatagttg
     6001 agtcgatact tcggcgatca ccgcttccct catgatgttt aactcctgaa ttaagccgcg
     6061 ccgcgaagcg gtgtcggctt gaatgaattg ttaggcgtca tcctgtgctc ccgagaacca
     6121 gtaccagtac atcgctgttt cgttcgagac ttgaggtcta gttttatacg tgaacaggtc
     6181 aatgccgccg agagtaaagc cacattttgc gtacaaattg caggcaggta cattgttcgt
     6241 ttgtgccatg atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacaca
     6301 acgtggcttt gttgaataaa tcgaactttt gctgagttga aggatcagat cacgcatctt
     6361 cccgacaacg cagaccgttc cgtggcaaag caaaagttca aaatcaccaa ctggtccacc
     6421 tacaacaaag ctctcatcaa ccgtggctcc ctcactttct ggctggatga tggggcgatt
     6481 caggagatcc ccatccaaca gcccgccgtc gagcgggctt ttttatcccc ggaagcctgt
     6541 ggatagaggg tagttatcca cgtgaaaccg ctaatgcccc gcaaagcctt gattcacggg
     6601 gctttccggc ccgctccaaa aactatccac gtgaaatcgc taatcagggt acgtgaaatc
     6661 gctaatcgga gtacgtgaaa tcgctaataa ggtcacgtga aatcgctaat caaaaaggca
     6721 cgtgagaacg ctaatagccc tttcagatca acagcttgca aacacccctc gctccggcaa
     6781 gtagttacag caagtagtat gttcaattag cttttcaatt atgaatatat atatcaatta
     6841 ttggtcgccc ttggcttgtg gacaatgcgc tacgcgcacc ggctccgccc gtggacaacc
     6901 gcaagcggtt gcccaccgtc gagcgccttt gcccacaacc cggcggccgg ccgcaacaga
     6961 tcgttttata aatttttttt tttgaaaaag aaaaagcccg aaaggcggca acctctcggg
     7021 cttctggatt tccgatcccc ggaattagac tct
//
