LOCUS       sequence-393465-        7242 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proUBQ10::PIP2A-mCherry-NOSt_proUBQ10::GFP-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7242)
  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 7242)
  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: JAB3423
FEATURES             Location/Qualifiers
     source          1..7242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    10..32
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     CDS             1555..2265
                     /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(1700..1718)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      2284..2536
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3243..3271)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3866..3886
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4257..4277)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4257..4275)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4296..4312)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4296..4312)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4309..4331)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4320..4336
                     /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(4344..4350,4351..4368,4369..4374))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4389..4410
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4527..4544)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4649..4673
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4764..5352)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4844..4863)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5443..6231)
                     /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      6689..7124
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 ttagactctt ggcaggatat attgtggtgt aacgttaagc tttggttgtc gacgagtcag
       61 taataaacgg cgtcaaagtg gttgcagccg gcacacacga gtcgtgttta tcaactcaaa
      121 gcacaaatac ttttcctcaa cctaaaaata aggcaattag ccaaaaacaa ctttgcgtgt
      181 aaacaacgct caatacacgt gtcattttat tattagctat tgcttcaccg ccttagcttt
      241 ctcgtgacct agtcgtcctc gtcttttctt cttcttcttc tataaaacaa tacccaaaga
      301 gctcttcttc ttcacaattc agatttcaat ttctcaaaat cttaaaaact ttctctcaat
      361 tctctctacc gtgatcaagg taaatttctg tgttccttat tctctcaaaa tcttcgattt
      421 tgttttcgtt cgatcccaat ttcgtatatg ttctttggtt tagattctgt taatcttaga
      481 tcgtagacga ttttctgggt ttgatcgtta gatatcatct taattctcga ttagggtttc
      541 atagatatca tccgatttgt tcaaataatt tgagttttgt cgaataatta ctcttcgatt
      601 tgtgatttct atctagatct ggtgttagtt tctagtttgt gcgatcgaat ttgtcgatta
      661 atctgagttt ttctgattaa cagcaccatg gcaaaggatg tggaagccgt tcccggagaa
      721 ggatttcaga caagagacta tcaagatccg ccaccagctc cgtttattga tggagcggag
      781 ctaaagaagt ggtctttcta cagagcagtt atcgcagagt tcgtagccac tctcctcttc
      841 ttatacatca ccgttttgac agtcatcggt tacaagattc agtccgatac tgatgccggt
      901 ggcgtagatt gcggcggagt tggaatcctc ggtatcgctt gggcctttgg tggtatgatc
      961 ttcatcctcg tctactgcac cgccggtatc tctggtggtc acattaaccc agcggtgaca
     1021 tttgggctat tcttggcacg taaagtgtcg ttacctaggg ccctattgta cataatcgct
     1081 cagtgtttgg gtgcgatttg tggagttggt tttgtcaaag ccttccaaag ctcttactac
     1141 acccgttacg gaggtggagc caactctcta gccgatggct acagcacagg gaccggtcta
     1201 gccgcagaga tcattggtac tttcgttctt gtctacaccg tattctctgc cactgacccc
     1261 aaacgtagtg ccagagactc ccacgttccg gtgttggcgc cacttccaat cggatttgcc
     1321 gtgttcatgg tacatttggc taccattccc attaccggaa ccggaattaa cccggcaagg
     1381 agtttcggag ctgccgtaat ctacaacaag agcaagccat gggatgacca ctggatattt
     1441 tgggttggac cattcattgg agctgcgata gctgcattct accaccaatt cgttctaaga
     1501 gcttcaggtt ctaagtctct tggatcattc agaagtgctg ccaacgtcgg atccatggtg
     1561 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
     1621 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     1681 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
     1741 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
     1801 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     1861 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     1921 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     1981 aataagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
     2041 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
     2101 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
     2161 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
     2221 gagggccgcc actccaccgg cggcatggag gagctgtaca agtaaagcgg ccgcccggct
     2281 gcagatcgtt caaacatttg gcaataaagt ttcttaagat tgaatcctgt tgccggtctt
     2341 gcgatgatta tcatataatt tctgttgaat tacgttaagc atgtaataat taacatgtaa
     2401 tgcatgacgt tatttatgag atgggttttt atgattagag tcccgcaatt atacatttaa
     2461 tacgcgatag aaaacaaaat atagcgcgca aactaggata aattatcgcg cgcggtgtca
     2521 tctatgttac tagatcgggg cgcgccctcg aggaagacgt aatgtggttg tcgacgagtc
     2581 agtaataaac ggcgtcaaag tggttgcagc cggcacacac gagtcgtgtt tatcaactca
     2641 aagcacaaat acttttcctc aacctaaaaa taaggcaatt agccaaaaac aactttgcgt
     2701 gtaaacaacg ctcaatacac gtgtcatttt attattagct attgcttcac cgccttagct
     2761 ttctcgtgac ctagtcgtcc tcgtcttttc ttcttcttct tctataaaac aatacccaaa
     2821 gagctcttct tcttcacaat tcagatttca atttctcaaa atcttaaaaa ctttctctca
     2881 attctctcta ccgtgatcaa ggtaaatttc tgtgttcctt attctctcaa aatcttcgat
     2941 tttgttttcg ttcgatccca atttcgtata tgttctttgg tttagattct gttaatctta
     3001 gatcgaagac gattttctgg gtttgatcgt tagatatcat cttaattctc gattagggtt
     3061 tcatagatat catccgattt gttcaaataa tttgagtttt gtcgaataat tactcttcga
     3121 tttgtgattt ctatctagat ctggtgttag tttctagttt gtgcgatcga atttgtcgat
     3181 taatctgagt ttttctgatt aacagcacca tggtgagtaa aggagaagaa cttttcactg
     3241 gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca
     3301 gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta
     3361 ctggaaaact acctgttcca tggccaaccc tggtcaccac cctgacctac ggcgtgcagt
     3421 gcttctcccg ttaccctgat catatgaagc ggcacgactt cttcaagagc gccatgcctg
     3481 agggatacgt gcaggagagg accatcttct tcaaggacga cgggaactac aagacacgtg
     3541 ctgaagtcaa gtttgaggga gacaccctcg tcaacaggat cgagcttaag ggaatcgatt
     3601 tcaaggagga cggaaacatc ctcggccaca agttggaata caactacaac tcccacaacg
     3661 tatacatcat ggccgacaag caaaagaacg gcatcaaagc caacttcaag acccgccaca
     3721 acatcgaaga cggcggcgtg caactcgctg atcattatca acaaaatact ccaattggcg
     3781 atggccctgt ccttttacca gacaaccatt acctgtccac acaatctgcc ctttcgaaag
     3841 atcccaacga aaagagagac cacatggtcc ttcttgagtt tgtaacagct gctgggatta
     3901 cacatggcat ggatgaacta tacaaataat gtcaagccat tctctcgcag atgatgttca
     3961 ctttgtgttt tacttccttt atgcattcac agcaataaaa gaaagaaatc tccatcgctt
     4021 ttggttttct tctctgtctt aagttagtcg ttttcgtgtc taatctatta cttatcattg
     4081 taatagactc ttcttctatt gagatttgaa tataaactaa aacacattcc attttactgt
     4141 gttctcaaca ttcagaatgc aaacggacta accgtagtga ctcggcttga gtcttcgggc
     4201 tagagcggac tagttctaga gcggccgcca ccgcggtgga gctccagctt ttgttccctt
     4261 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4321 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4381 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4441 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4501 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4561 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4621 gataacgcag gaaagaacat gaaggccttg acaggatata ttggcgggta aactaagtcg
     4681 ctgtatgtgt ttgtttgaga tctcatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4741 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4801 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4861 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4921 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4981 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5041 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5101 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5161 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5221 tgtgctctgc tgaagccagt taccttcgga agaagagttg gtagctcttg atccggcaaa
     5281 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5341 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5401 aactcacgtt aagggatttt ggtcatgaat tcttgttaga cattatttgc cgactacctt
     5461 ggtgatctcg cctttcacgt agtgaacaaa ttcttccaac tgatctgcgc gcgaggccaa
     5521 gcgatcttct tgtccaagat aagcctgcct agcttcaagt atgacgggct gatactgggc
     5581 cggcaggcgc tccattgccc agtcggcagc gacatccttc ggcgcgattt tgccggttac
     5641 tgcgctgtac caaatgcggg acaacgtaag cactacattt cgctcatcgc cagcccagtc
     5701 gggcggcgag ttccatagcg ttaaggtttc atttagcgcc tcaaatagat cctgttcagg
     5761 aaccggatca aagagttcct ccgccgctgg acctaccaag gcaacgctat gttctcttgc
     5821 ttttgtcagc aagatagcca gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag
     5881 aatgtcattg cgctgccatt ctccaaattg cagttcgcgc ttagctggat aacgccacgg
     5941 aatgatgtcg tcgtgcacaa caatggtgac ttctacagcg cggagaatct cgctctctcc
     6001 aggggaagcc gaagtttcca aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag
     6061 ccttacggtc accgtaacca gcaaatcaat atcactgtgt ggcttcaggc cgccatccac
     6121 tgcggagccg tacaaatgta cggccagcaa cgtcggttcg agatggcgct cgatgacgcc
     6181 aactacctct gatagttgag tcgatacttc ggcgatcacc gcttccctca tgatgtttaa
     6241 ctcctgaatt aagccgcgcc gcgaagcggt gtcggcttga atgaattgtt aggcgtcatc
     6301 ctgtgctccc gagaaccagt accagtacat cgctgtttcg ttcgagactt gaggtctagt
     6361 tttatacgtg aacaggtcaa tgccgccgag agtaaagcca cattttgcgt acaaattgca
     6421 ggcaggtaca ttgttcgttt gtgccatgat gatatatttt tatcttgtgc aatgtaacat
     6481 cagagatttt gagacacaac gtggctttgt tgaataaatc gaacttttgc tgagttgaag
     6541 gatcagatca cgcatcttcc cgacaacgca gaccgttccg tggcaaagca aaagttcaaa
     6601 atcaccaact ggtccaccta caacaaagct ctcatcaacc gtggctccct cactttctgg
     6661 ctggatgatg gggcgattca ggagatcccc atccaacagc ccgccgtcga gcgggctttt
     6721 ttatccccgg aagcctgtgg atagagggta gttatccacg tgaaaccgct aatgccccgc
     6781 aaagccttga ttcacggggc tttccggccc gctccaaaaa ctatccacgt gaaatcgcta
     6841 atcagggtac gtgaaatcgc taatcggagt acgtgaaatc gctaataagg tcacgtgaaa
     6901 tcgctaatca aaaaggcacg tgagaacgct aatagccctt tcagatcaac agcttgcaaa
     6961 cacccctcgc tccggcaagt agttacagca agtagtatgt tcaattagct tttcaattat
     7021 gaatatatat atcaattatt ggtcgccctt ggcttgtgga caatgcgcta cgcgcaccgg
     7081 ctccgcccgt ggacaaccgc aagcggttgc ccaccgtcga gcgcctttgc ccacaacccg
     7141 gcggccggcc gcaacagatc gttttataaa tttttttttt tgaaaaagaa aaagcccgaa
     7201 aggcggcaac ctctcgggct tctggatttc cgatccccgg aa
//
