LOCUS       sequence-393618-        7647 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proUBQ10::PIP2A-mCherry-NOSt_proT::GFP-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7647)
  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 7647)
  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: JAB3424
FEATURES             Location/Qualifiers
     source          1..7647
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    19..41
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     CDS             1564..2274
                     /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(1709..1727)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      2293..2545
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3468..3496)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          3636..3824
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     4280..4300
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4671..4691)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4671..4689)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4710..4726)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4710..4726)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4723..4745)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4734..4750
                     /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(4758..4764,4765..4782,4783..4788))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4803..4824
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4941..4958)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5063..5087
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5178..5766)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5258..5277)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5857..6645)
                     /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      7103..7538
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 tccccggaat tagactcttg gcaggatata ttgtggtgta acgttaagct ttggttgtcg
       61 acgagtcagt aataaacggc gtcaaagtgg ttgcagccgg cacacacgag tcgtgtttat
      121 caactcaaag cacaaatact tttcctcaac ctaaaaataa ggcaattagc caaaaacaac
      181 tttgcgtgta aacaacgctc aatacacgtg tcattttatt attagctatt gcttcaccgc
      241 cttagctttc tcgtgaccta gtcgtcctcg tcttttcttc ttcttcttct ataaaacaat
      301 acccaaagag ctcttcttct tcacaattca gatttcaatt tctcaaaatc ttaaaaactt
      361 tctctcaatt ctctctaccg tgatcaaggt aaatttctgt gttccttatt ctctcaaaat
      421 cttcgatttt gttttcgttc gatcccaatt tcgtatatgt tctttggttt agattctgtt
      481 aatcttagat cgtagacgat tttctgggtt tgatcgttag atatcatctt aattctcgat
      541 tagggtttca tagatatcat ccgatttgtt caaataattt gagttttgtc gaataattac
      601 tcttcgattt gtgatttcta tctagatctg gtgttagttt ctagtttgtg cgatcgaatt
      661 tgtcgattaa tctgagtttt tctgattaac agcaccatgg caaaggatgt ggaagccgtt
      721 cccggagaag gatttcagac aagagactat caagatccgc caccagctcc gtttattgat
      781 ggagcggagc taaagaagtg gtctttctac agagcagtta tcgcagagtt cgtagccact
      841 ctcctcttct tatacatcac cgttttgaca gtcatcggtt acaagattca gtccgatact
      901 gatgccggtg gcgtagattg cggcggagtt ggaatcctcg gtatcgcttg ggcctttggt
      961 ggtatgatct tcatcctcgt ctactgcacc gccggtatct ctggtggtca cattaaccca
     1021 gcggtgacat ttgggctatt cttggcacgt aaagtgtcgt tacctagggc cctattgtac
     1081 ataatcgctc agtgtttggg tgcgatttgt ggagttggtt ttgtcaaagc cttccaaagc
     1141 tcttactaca cccgttacgg aggtggagcc aactctctag ccgatggcta cagcacaggg
     1201 accggtctag ccgcagagat cattggtact ttcgttcttg tctacaccgt attctctgcc
     1261 actgacccca aacgtagtgc cagagactcc cacgttccgg tgttggcgcc acttccaatc
     1321 ggatttgccg tgttcatggt acatttggct accattccca ttaccggaac cggaattaac
     1381 ccggcaagga gtttcggagc tgccgtaatc tacaacaaga gcaagccatg ggatgaccac
     1441 tggatatttt gggttggacc attcattgga gctgcgatag ctgcattcta ccaccaattc
     1501 gttctaagag cttcaggttc taagtctctt ggatcattca gaagtgctgc caacgtcgga
     1561 tccatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
     1621 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     1681 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
     1741 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
     1801 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     1861 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     1921 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     1981 gtaatgcaga ataagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     2041 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     2101 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     2161 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     2221 gaacgcgccg agggccgcca ctccaccggc ggcatggagg agctgtacaa gtaaagcggc
     2281 cgcccggctg cagatcgttc aaacatttgg caataaagtt tcttaagatt gaatcctgtt
     2341 gccggtcttg cgatgattat catataattt ctgttgaatt acgttaagca tgtaataatt
     2401 aacatgtaat gcatgacgtt atttatgaga tgggttttta tgattagagt cccgcaatta
     2461 tacatttaat acgcgataga aaacaaaata tagcgcgcaa actaggataa attatcgcgc
     2521 gcggtgtcat ctatgttact agatcggggc gcgccctcga ggaagacgta atgggtcccc
     2581 agatttgcct tttcaatttc agaaagaatg ctaacccaca gatggttaga gaggcttacg
     2641 cagcagctct catcaagacg atctacccga gcaataatct ccaggaaatc aaataccttc
     2701 ccaagaaggt taaagatgca gtcaaaagat tcaggactaa ctgcatcaag aacacagaga
     2761 aagatatatt tctcaagatc agaagtacta ttccagtatg gacgattcaa ggcttgcttc
     2821 acaaaccaag gcaagtaata gagattggag tctctaaaaa ggtagttccc actgaatcaa
     2881 aggccatgga gtcaaagatt caaatagagg acctaacaga actcgccgta aagactggcg
     2941 aacagttcat acagagtctc ttacgactca atgacaagaa gaaaatcttc gtcaacatgg
     3001 tggagcacga cacacttgtc tactccaaaa atatcaaaga tacagtctca gaagaccaaa
     3061 gggcaattga gacttttcaa caaagggtaa tatccggaaa cctcctcgga ttccattgcc
     3121 cagctatctg tcactttatt gtgaagatag tggaaaagga aggtggctcc tacaaatgcc
     3181 atcattgcga taaaggaaag gccatcgttg aagatgcctc tgccgacagt ggtcccaaag
     3241 atggaccccc acccacgagg agcatcgtgg aaaaagaaga cgttccaacc acgtcttcaa
     3301 agcaagtgga ttgatgtgat atctccactg acgtaaggga tgacgcacaa tcccactatc
     3361 cttcgcaaga tagataattc tatataaact ctatcactga tagagtggag agaacacggg
     3421 ggactctaga caccatggtg agtaaaggag aagaactttt cactggagtt gtcccaattc
     3481 ttgttgaatt agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag
     3541 gtgatgcaac atacggaaaa cttaccctta aatttatttg cactactgga aaactacctg
     3601 ttccatggcc aaccctggtc accaccctga cctacgtaag tttctgcttc tacctttgat
     3661 atatatataa taattatcat taattagtag taatataata tttcaaatat ttttttcaaa
     3721 ataaaagaat gtagtatata gcaattgctt ttctgtagtt tataagtgtg tatattttaa
     3781 tttataactt ttctaatata tgaccaaaat ttgttgatgt gcagggcgtg cagtgcttct
     3841 cccgttaccc tgatcatatg aagcggcacg acttcttcaa gagcgccatg cctgagggat
     3901 acgtgcagga gaggaccatc ttcttcaagg acgacgggaa ctacaagaca cgtgctgaag
     3961 tcaagtttga gggtgacacc ctcgtcaaca ggatcgagct taagggaatc gatttcaagg
     4021 aggacggaaa catcctcggc cacaagttgg aatacaacta caactcccac aacgtataca
     4081 tcatggccga caagcaaaag aacggcatca aagccaactt caagacccgc cacaacatcg
     4141 aggacggcgg cgtgcaactc gctgatcatt atcaacaaaa tactccaatt ggcgatggcc
     4201 ctgtcctttt accagacaac cattacctgt ccacacaatc tgccctttcg aaagatccca
     4261 acgaaaagag agatcacatg gtccttcttg agtttgtaac agctgctggg attacacatg
     4321 gcatggatga actatacaaa taatgtcaag ccattctctc gcagatgatg ttcactttgt
     4381 gttttacttc ctttatgcat tcacagcaat aaaagaaaga aatctccatc gcttttggtt
     4441 ttcttctctg tcttaagtta gtcgttttcg tgtctaatct attacttatc attgtaatag
     4501 actcttcttc tattgagatt tgaatataaa ctaaaacaca ttccatttta ctgtgttctc
     4561 aacattcaga atgcaaacgg actaaccgta gtgactcggc ttgagtcttc gggctagagc
     4621 ggactagttc tagagcggcc gccaccgcgg tggagctcca gcttttgttc cctttagtga
     4681 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4741 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4801 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4861 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4921 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4981 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5041 gcaggaaaga acatgaaggc cttgacagga tatattggcg ggtaaactaa gtcgctgtat
     5101 gtgtttgttt gagatctcat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5161 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5221 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5281 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5341 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5401 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5461 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5521 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5581 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgtgct
     5641 ctgctgaagc cagttacctt cggaagaaga gttggtagct cttgatccgg caaacaaacc
     5701 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5761 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5821 cgttaaggga ttttggtcat gaattcttgt tagacattat ttgccgacta ccttggtgat
     5881 ctcgcctttc acgtagtgaa caaattcttc caactgatct gcgcgcgagg ccaagcgatc
     5941 ttcttgtcca agataagcct gcctagcttc aagtatgacg ggctgatact gggccggcag
     6001 gcgctccatt gcccagtcgg cagcgacatc cttcggcgcg attttgccgg ttactgcgct
     6061 gtaccaaatg cgggacaacg taagcactac atttcgctca tcgccagccc agtcgggcgg
     6121 cgagttccat agcgttaagg tttcatttag cgcctcaaat agatcctgtt caggaaccgg
     6181 atcaaagagt tcctccgccg ctggacctac caaggcaacg ctatgttctc ttgcttttgt
     6241 cagcaagata gccagatcaa tgtcgatcgt ggctggctcg aagatacctg caagaatgtc
     6301 attgcgctgc cattctccaa attgcagttc gcgcttagct ggataacgcc acggaatgat
     6361 gtcgtcgtgc acaacaatgg tgacttctac agcgcggaga atctcgctct ctccagggga
     6421 agccgaagtt tccaaaaggt cgttgatcaa agctcgccgc gttgtttcat caagccttac
     6481 ggtcaccgta accagcaaat caatatcact gtgtggcttc aggccgccat ccactgcgga
     6541 gccgtacaaa tgtacggcca gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac
     6601 ctctgatagt tgagtcgata cttcggcgat caccgcttcc ctcatgatgt ttaactcctg
     6661 aattaagccg cgccgcgaag cggtgtcggc ttgaatgaat tgttaggcgt catcctgtgc
     6721 tcccgagaac cagtaccagt acatcgctgt ttcgttcgag acttgaggtc tagttttata
     6781 cgtgaacagg tcaatgccgc cgagagtaaa gccacatttt gcgtacaaat tgcaggcagg
     6841 tacattgttc gtttgtgcca tgatgatata tttttatctt gtgcaatgta acatcagaga
     6901 ttttgagaca caacgtggct ttgttgaata aatcgaactt ttgctgagtt gaaggatcag
     6961 atcacgcatc ttcccgacaa cgcagaccgt tccgtggcaa agcaaaagtt caaaatcacc
     7021 aactggtcca cctacaacaa agctctcatc aaccgtggct ccctcacttt ctggctggat
     7081 gatggggcga ttcaggagat ccccatccaa cagcccgccg tcgagcgggc ttttttatcc
     7141 ccggaagcct gtggatagag ggtagttatc cacgtgaaac cgctaatgcc ccgcaaagcc
     7201 ttgattcacg gggctttccg gcccgctcca aaaactatcc acgtgaaatc gctaatcagg
     7261 gtacgtgaaa tcgctaatcg gagtacgtga aatcgctaat aaggtcacgt gaaatcgcta
     7321 atcaaaaagg cacgtgagaa cgctaatagc cctttcagat caacagcttg caaacacccc
     7381 tcgctccggc aagtagttac agcaagtagt atgttcaatt agcttttcaa ttatgaatat
     7441 atatatcaat tattggtcgc ccttggcttg tggacaatgc gctacgcgca ccggctccgc
     7501 ccgtggacaa ccgcaagcgg ttgcccaccg tcgagcgcct ttgcccacaa cccggcggcc
     7561 ggccgcaaca gatcgtttta taaatttttt tttttgaaaa agaaaaagcc cgaaaggcgg
     7621 caacctctcg ggcttctgga tttccga
//
