LOCUS       sequence-391158-        7647 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proUBQ10::PIP2A-mCherry-NOSt_proTT*::GFPint_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: JAB3236
FEATURES             Location/Qualifiers
     source          1..7647
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    28..50
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     CDS             1573..2283
                     /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(1718..1736)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      2302..2554
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3477..3505)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          3645..3833
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     4289..4309
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4680..4700)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4680..4698)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4719..4735)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4719..4735)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4732..4754)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4743..4759
                     /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(4767..4773,4774..4791,4792..4797))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4812..4833
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4950..4967)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5072..5096
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5187..5775)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5267..5286)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5866..6654)
                     /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      7112..7547
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 gatttccgat ccccggaatt agactcttgg caggatatat tgtggtgtaa cgttaagctt
       61 tggttgtcga cgagtcagta ataaacggcg tcaaagtggt tgcagccggc acacacgagt
      121 cgtgtttatc aactcaaagc acaaatactt ttcctcaacc taaaaataag gcaattagcc
      181 aaaaacaact ttgcgtgtaa acaacgctca atacacgtgt cattttatta ttagctattg
      241 cttcaccgcc ttagctttct cgtgacctag tcgtcctcgt cttttcttct tcttcttcta
      301 taaaacaata cccaaagagc tcttcttctt cacaattcag atttcaattt ctcaaaatct
      361 taaaaacttt ctctcaattc tctctaccgt gatcaaggta aatttctgtg ttccttattc
      421 tctcaaaatc ttcgattttg ttttcgttcg atcccaattt cgtatatgtt ctttggttta
      481 gattctgtta atcttagatc gtagacgatt ttctgggttt gatcgttaga tatcatctta
      541 attctcgatt agggtttcat agatatcatc cgatttgttc aaataatttg agttttgtcg
      601 aataattact cttcgatttg tgatttctat ctagatctgg tgttagtttc tagtttgtgc
      661 gatcgaattt gtcgattaat ctgagttttt ctgattaaca gcaccatggc aaaggatgtg
      721 gaagccgttc ccggagaagg atttcagaca agagactatc aagatccgcc accagctccg
      781 tttattgatg gagcggagct aaagaagtgg tctttctaca gagcagttat cgcagagttc
      841 gtagccactc tcctcttctt atacatcacc gttttgacag tcatcggtta caagattcag
      901 tccgatactg atgccggtgg cgtagattgc ggcggagttg gaatcctcgg tatcgcttgg
      961 gcctttggtg gtatgatctt catcctcgtc tactgcaccg ccggtatctc tggtggtcac
     1021 attaacccag cggtgacatt tgggctattc ttggcacgta aagtgtcgtt acctagggcc
     1081 ctattgtaca taatcgctca gtgtttgggt gcgatttgtg gagttggttt tgtcaaagcc
     1141 ttccaaagct cttactacac ccgttacgga ggtggagcca actctctagc cgatggctac
     1201 agcacaggga ccggtctagc cgcagagatc attggtactt tcgttcttgt ctacaccgta
     1261 ttctctgcca ctgaccccaa acgtagtgcc agagactccc acgttccggt gttggcgcca
     1321 cttccaatcg gatttgccgt gttcatggta catttggcta ccattcccat taccggaacc
     1381 ggaattaacc cggcaaggag tttcggagct gccgtaatct acaacaagag caagccatgg
     1441 gatgaccact ggatattttg ggttggacca ttcattggag ctgcgatagc tgcattctac
     1501 caccaattcg ttctaagagc ttcaggttct aagtctcttg gatcattcag aagtgctgcc
     1561 aacgtcggat ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
     1621 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
     1681 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
     1741 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
     1801 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
     1861 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
     1921 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     1981 gacggccccg taatgcagaa taagaccatg ggctgggagg cctcctccga gcggatgtac
     2041 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     2101 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     2161 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     2221 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggagga gctgtacaag
     2281 taaagcggcc gcccggctgc agatcgttca aacatttggc aataaagttt cttaagattg
     2341 aatcctgttg ccggtcttgc gatgattatc atataatttc tgttgaatta cgttaagcat
     2401 gtaataatta acatgtaatg catgacgtta tttatgagat gggtttttat gattagagtc
     2461 ccgcaattat acatttaata cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa
     2521 ttatcgcgcg cggtgtcatc tatgttacta gatcggggcg cgccctcgag gaagacgtaa
     2581 tgggtcccca gatttgcctt ttcaatttca gaaagaatgc taacccacag atggttagag
     2641 aggcttacgc agcagctctc atcaagacga tctacccgag caataatctc caggaaatca
     2701 aataccttcc caagaaggtt aaagatgcag tcaaaagatt caggactaac tgcatcaaga
     2761 acacagagaa agatatattt ctcaagatca gaagtactat tccagtatgg acgattcaag
     2821 gcttgcttca caaaccaagg caagtaatag agattggagt ctctaaaaag gtagttccca
     2881 ctgaatcaaa ggccatggag tcaaagattc aaatagagga cctaacagaa ctcgccgtaa
     2941 agactggcga acagttcata cagagtctct tacgactcaa tgacaagaag aaaatcttcg
     3001 tcaacatggt ggagcacgac acacttgtct actccaaaaa tatcaaagat acagtctcag
     3061 aagaccaaag ggcaattgag acttttcaac aaagggtaat atccggaaac ctcctcggat
     3121 tccattgccc agctatctgt cactttattg tgaagatagt ggaaaaggaa ggtggctcct
     3181 acaaatgcca tcattgcgat aaaggaaagg ccatcgttga agatgcctct gccgacagtg
     3241 gtcccaaaga tggaccccca cccacgagga gcatcgtgga aaaagaagac gttccaacca
     3301 cgtcttcaaa gcaagtggat tgatgtgata tctccactga cgtaagggat gacgcacaat
     3361 cccactatcc ttcgcaagat agataattct atataaactc tatcactgat agagtggact
     3421 ctatcactga tagagtagac accatggtga gtaaaggaga agaacttttc actggagttg
     3481 tcccaattct tgttgaatta gatggtgatg ttaatgggca caaattttct gtcagtggag
     3541 agggtgaagg tgatgcaaca tacggaaaac ttacccttaa atttatttgc actactggaa
     3601 aactacctgt tccatggcca accctggtca ccaccctgac ctacgtaagt ttctgcttct
     3661 acctttgata tatatataat aattatcatt aattagtagt aatataatat ttcaaatatt
     3721 tttttcaaaa taaaagaatg tagtatatag caattgcttt tctgtagttt ataagtgtgt
     3781 atattttaat ttataacttt tctaatatat gaccaaaatt tgttgatgtg cagggcgtgc
     3841 agtgcttctc ccgttaccct gatcatatga agcggcacga cttcttcaag agcgccatgc
     3901 ctgagggata cgtgcaggag aggaccatct tcttcaagga cgacgggaac tacaagacac
     3961 gtgctgaagt caagtttgag ggtgacaccc tcgtcaacag gatcgagctt aagggaatcg
     4021 atttcaagga ggacggaaac atcctcggcc acaagttgga atacaactac aactcccaca
     4081 acgtatacat catggccgac aagcaaaaga acggcatcaa agccaacttc aagacccgcc
     4141 acaacatcga agacggcggc gtgcaactcg ctgatcatta tcaacaaaat actccaattg
     4201 gcgatggccc tgtcctttta ccagacaacc attacctgtc cacacaatct gccctttcga
     4261 aagatcccaa cgaaaagaga gatcacatgg tccttcttga gtttgtaaca gctgctggga
     4321 ttacacatgg catggatgaa ctatacaaat aatgtcaagc cattctctcg cagatgatgt
     4381 tcactttgtg ttttacttcc tttatgcatt cacagcaata aaagaaagaa atctccatcg
     4441 cttttggttt tcttctctgt cttaagttag tcgttttcgt gtctaatcta ttacttatca
     4501 ttgtaataga ctcttcttct attgagattt gaatataaac taaaacacat tccattttac
     4561 tgtgttctca acattcagaa tgcaaacgga ctaaccgtag tgactcggct tgagtcttcg
     4621 ggctagagcg gactagttct agagcggccg ccaccgcggt ggagctccag cttttgttcc
     4681 ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
     4741 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4801 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4861 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4921 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4981 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5041 ggggataacg caggaaagaa catgaaggcc ttgacaggat atattggcgg gtaaactaag
     5101 tcgctgtatg tgtttgtttg agatctcatg tgagcaaaag gccagcaaaa ggccaggaac
     5161 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5221 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5281 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5341 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5401 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5461 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5521 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5581 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5641 atctgtgctc tgctgaagcc agttaccttc ggaagaagag ttggtagctc ttgatccggc
     5701 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5761 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5821 gaaaactcac gttaagggat tttggtcatg aattcttgtt agacattatt tgccgactac
     5881 cttggtgatc tcgcctttca cgtagtgaac aaattcttcc aactgatctg cgcgcgaggc
     5941 caagcgatct tcttgtccaa gataagcctg cctagcttca agtatgacgg gctgatactg
     6001 ggccggcagg cgctccattg cccagtcggc agcgacatcc ttcggcgcga ttttgccggt
     6061 tactgcgctg taccaaatgc gggacaacgt aagcactaca tttcgctcat cgccagccca
     6121 gtcgggcggc gagttccata gcgttaaggt ttcatttagc gcctcaaata gatcctgttc
     6181 aggaaccgga tcaaagagtt cctccgccgc tggacctacc aaggcaacgc tatgttctct
     6241 tgcttttgtc agcaagatag ccagatcaat gtcgatcgtg gctggctcga agatacctgc
     6301 aagaatgtca ttgcgctgcc attctccaaa ttgcagttcg cgcttagctg gataacgcca
     6361 cggaatgatg tcgtcgtgca caacaatggt gacttctaca gcgcggagaa tctcgctctc
     6421 tccaggggaa gccgaagttt ccaaaaggtc gttgatcaaa gctcgccgcg ttgtttcatc
     6481 aagccttacg gtcaccgtaa ccagcaaatc aatatcactg tgtggcttca ggccgccatc
     6541 cactgcggag ccgtacaaat gtacggccag caacgtcggt tcgagatggc gctcgatgac
     6601 gccaactacc tctgatagtt gagtcgatac ttcggcgatc accgcttccc tcatgatgtt
     6661 taactcctga attaagccgc gccgcgaagc ggtgtcggct tgaatgaatt gttaggcgtc
     6721 atcctgtgct cccgagaacc agtaccagta catcgctgtt tcgttcgaga cttgaggtct
     6781 agttttatac gtgaacaggt caatgccgcc gagagtaaag ccacattttg cgtacaaatt
     6841 gcaggcaggt acattgttcg tttgtgccat gatgatatat ttttatcttg tgcaatgtaa
     6901 catcagagat tttgagacac aacgtggctt tgttgaataa atcgaacttt tgctgagttg
     6961 aaggatcaga tcacgcatct tcccgacaac gcagaccgtt ccgtggcaaa gcaaaagttc
     7021 aaaatcacca actggtccac ctacaacaaa gctctcatca accgtggctc cctcactttc
     7081 tggctggatg atggggcgat tcaggagatc cccatccaac agcccgccgt cgagcgggct
     7141 tttttatccc cggaagcctg tggatagagg gtagttatcc acgtgaaacc gctaatgccc
     7201 cgcaaagcct tgattcacgg ggctttccgg cccgctccaa aaactatcca cgtgaaatcg
     7261 ctaatcaggg tacgtgaaat cgctaatcgg agtacgtgaa atcgctaata aggtcacgtg
     7321 aaatcgctaa tcaaaaaggc acgtgagaac gctaatagcc ctttcagatc aacagcttgc
     7381 aaacacccct cgctccggca agtagttaca gcaagtagta tgttcaatta gcttttcaat
     7441 tatgaatata tatatcaatt attggtcgcc cttggcttgt ggacaatgcg ctacgcgcac
     7501 cggctccgcc cgtggacaac cgcaagcggt tgcccaccgt cgagcgcctt tgcccacaac
     7561 ccggcggccg gccgcaacag atcgttttat aaattttttt ttttgaaaaa gaaaaagccc
     7621 gaaaggcggc aacctctcgg gcttctg
//
