LOCUS       sequence-391303-        7049 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proUBQ10::PIP2A-mCherry-NOSt_4XPhlFO::GFPint-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7049)
  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 7049)
  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: JAB3603
FEATURES             Location/Qualifiers
     source          1..7049
                     /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        2752..2798
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     primer_bind     complement(2852..2880)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          3020..3208
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     3664..3684
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4055..4075)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4055..4073)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4094..4110)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4094..4110)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4107..4129)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4118..4134
                     /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(4142..4148,4149..4166,4167..4172))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4187..4208
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4325..4342)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4447..4471
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4562..5150)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4642..4661)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5241..6029)
                     /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      6487..6922
                     /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 aatgatacga aacgtaccgt atcgttaagg tcaatgatac gaaacgtacc gtatcgttaa
     2641 ggtgtatgat acgaaacgta ccgtatcgtt aaggtagatg atacgaaacg taccgtatcg
     2701 ttaaggtcgg gtctaagcta gctaggccac ccacaatccc actatccttt cgcaagaccc
     2761 ttcctctata taaggaagtt catttcattt ggagagaaca cgggggactc tagacaccat
     2821 ggtgagtaaa ggagaagaac ttttcactgg agttgtccca attcttgttg aattagatgg
     2881 tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg caacatacgg
     2941 aaaacttacc cttaaattta tttgcactac tggaaaacta cctgttccat ggccaaccct
     3001 ggtcaccacc ctgacctacg taagtttctg cttctacctt tgatatatat ataataatta
     3061 tcattaatta gtagtaatat aatatttcaa atattttttt caaaataaaa gaatgtagta
     3121 tatagcaatt gcttttctgt agtttataag tgtgtatatt ttaatttata acttttctaa
     3181 tatatgacca aaatttgttg atgtgcaggg cgtgcagtgc ttctcccgtt accctgatca
     3241 tatgaagcgg cacgacttct tcaagagcgc catgcctgag ggatacgtgc aggagaggac
     3301 catcttcttc aaggacgacg ggaactacaa gacacgtgct gaagtcaagt ttgagggtga
     3361 caccctcgtc aacaggatcg agcttaaggg aatcgatttc aaggaggacg gaaacatcct
     3421 cggccacaag ttggaataca actacaactc ccacaacgta tacatcatgg ccgacaagca
     3481 aaagaacggc atcaaagcca acttcaagac ccgccacaac atcgaggacg gcggcgtgca
     3541 actcgctgat cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga
     3601 caaccattac ctgtccacac aatctgccct ttcgaaagat cccaacgaaa agagagatca
     3661 catggtcctt cttgagtttg taacagctgc tgggattaca catggcatgg atgaactata
     3721 caaataatgt caagccattc tctcgcagat gatgttcact ttgtgtttta cttcctttat
     3781 gcattcacag caataaaaga aagaaatctc catcgctttt ggttttcttc tctgtcttaa
     3841 gttagtcgtt ttcgtgtcta atctattact tatcattgta atagactctt cttctattga
     3901 gatttgaata taaactaaaa cacattccat tttactgtgt tctcaacatt cagaatgcaa
     3961 acggactaac cgtagtgact cggcttgagt cttcgggcta gagcggacta gttctagagc
     4021 ggccgccacc gcggtggagc tccagctttt gttcccttta gtgagggtta attgcgcgct
     4081 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     4141 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     4201 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     4261 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     4321 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     4381 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatga
     4441 aggccttgac aggatatatt ggcgggtaaa ctaagtcgct gtatgtgttt gtttgagatc
     4501 tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4561 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4621 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4681 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4741 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4801 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4861 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4921 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4981 ctaactacgg ctacactaga agaacagtat ttggtatctg tgctctgctg aagccagtta
     5041 ccttcggaag aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5101 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5161 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5221 tcatgaattc ttgttagaca ttatttgccg actaccttgg tgatctcgcc tttcacgtag
     5281 tgaacaaatt cttccaactg atctgcgcgc gaggccaagc gatcttcttg tccaagataa
     5341 gcctgcctag cttcaagtat gacgggctga tactgggccg gcaggcgctc cattgcccag
     5401 tcggcagcga catccttcgg cgcgattttg ccggttactg cgctgtacca aatgcgggac
     5461 aacgtaagca ctacatttcg ctcatcgcca gcccagtcgg gcggcgagtt ccatagcgtt
     5521 aaggtttcat ttagcgcctc aaatagatcc tgttcaggaa ccggatcaaa gagttcctcc
     5581 gccgctggac ctaccaaggc aacgctatgt tctcttgctt ttgtcagcaa gatagccaga
     5641 tcaatgtcga tcgtggctgg ctcgaagata cctgcaagaa tgtcattgcg ctgccattct
     5701 ccaaattgca gttcgcgctt agctggataa cgccacggaa tgatgtcgtc gtgcacaaca
     5761 atggtgactt ctacagcgcg gagaatctcg ctctctccag gggaagccga agtttccaaa
     5821 aggtcgttga tcaaagctcg ccgcgttgtt tcatcaagcc ttacggtcac cgtaaccagc
     5881 aaatcaatat cactgtgtgg cttcaggccg ccatccactg cggagccgta caaatgtacg
     5941 gccagcaacg tcggttcgag atggcgctcg atgacgccaa ctacctctga tagttgagtc
     6001 gatacttcgg cgatcaccgc ttccctcatg atgtttaact cctgaattaa gccgcgccgc
     6061 gaagcggtgt cggcttgaat gaattgttag gcgtcatcct gtgctcccga gaaccagtac
     6121 cagtacatcg ctgtttcgtt cgagacttga ggtctagttt tatacgtgaa caggtcaatg
     6181 ccgccgagag taaagccaca ttttgcgtac aaattgcagg caggtacatt gttcgtttgt
     6241 gccatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga gacacaacgt
     6301 ggctttgttg aataaatcga acttttgctg agttgaagga tcagatcacg catcttcccg
     6361 acaacgcaga ccgttccgtg gcaaagcaaa agttcaaaat caccaactgg tccacctaca
     6421 acaaagctct catcaaccgt ggctccctca ctttctggct ggatgatggg gcgattcagg
     6481 agatccccat ccaacagccc gccgtcgagc gggctttttt atccccggaa gcctgtggat
     6541 agagggtagt tatccacgtg aaaccgctaa tgccccgcaa agccttgatt cacggggctt
     6601 tccggcccgc tccaaaaact atccacgtga aatcgctaat cagggtacgt gaaatcgcta
     6661 atcggagtac gtgaaatcgc taataaggtc acgtgaaatc gctaatcaaa aaggcacgtg
     6721 agaacgctaa tagccctttc agatcaacag cttgcaaaca cccctcgctc cggcaagtag
     6781 ttacagcaag tagtatgttc aattagcttt tcaattatga atatatatat caattattgg
     6841 tcgcccttgg cttgtggaca atgcgctacg cgcaccggct ccgcccgtgg acaaccgcaa
     6901 gcggttgccc accgtcgagc gcctttgccc acaacccggc ggccggccgc aacagatcgt
     6961 tttataaatt tttttttttg aaaaagaaaa agcccgaaag gcggcaacct ctcgggcttc
     7021 tggatttccg atccccggaa ttagactct
//
