LOCUS       sequence-393423-        7700 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants 
            (proUBQ10::PIP2A-mCherry-NOSt_pro35S-BetIO::GFPint-NLS_ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7700)
  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 7700)
  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: JAB2500
FEATURES             Location/Qualifiers
     source          1..7700
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    8..30
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     CDS             1553..2263
                     /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(1698..1716)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      2282..2534
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        3057..3402
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     3345..3365
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     complement(3480..3508)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          3648..3836
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     4292..4312
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             4353..4373
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4713..4733)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4713..4731)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4752..4768)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4752..4768)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4765..4787)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4776..4792
                     /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(4800..4806,4807..4824,4825..4830))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4845..4866
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4983..5000)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5105..5129
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5220..5808)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5300..5319)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5899..6687)
                     /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      7145..7580
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 agactcttgg caggatatat tgtggtgtaa cgttaagctt tggttgtcga cgagtcagta
       61 ataaacggcg tcaaagtggt tgcagccggc acacacgagt cgtgtttatc aactcaaagc
      121 acaaatactt ttcctcaacc taaaaataag gcaattagcc aaaaacaact ttgcgtgtaa
      181 acaacgctca atacacgtgt cattttatta ttagctattg cttcaccgcc ttagctttct
      241 cgtgacctag tcgtcctcgt cttttcttct tcttcttcta taaaacaata cccaaagagc
      301 tcttcttctt cacaattcag atttcaattt ctcaaaatct taaaaacttt ctctcaattc
      361 tctctaccgt gatcaaggta aatttctgtg ttccttattc tctcaaaatc ttcgattttg
      421 ttttcgttcg atcccaattt cgtatatgtt ctttggttta gattctgtta atcttagatc
      481 gtagacgatt ttctgggttt gatcgttaga tatcatctta attctcgatt agggtttcat
      541 agatatcatc cgatttgttc aaataatttg agttttgtcg aataattact cttcgatttg
      601 tgatttctat ctagatctgg tgttagtttc tagtttgtgc gatcgaattt gtcgattaat
      661 ctgagttttt ctgattaaca gcaccatggc aaaggatgtg gaagccgttc ccggagaagg
      721 atttcagaca agagactatc aagatccgcc accagctccg tttattgatg gagcggagct
      781 aaagaagtgg tctttctaca gagcagttat cgcagagttc gtagccactc tcctcttctt
      841 atacatcacc gttttgacag tcatcggtta caagattcag tccgatactg atgccggtgg
      901 cgtagattgc ggcggagttg gaatcctcgg tatcgcttgg gcctttggtg gtatgatctt
      961 catcctcgtc tactgcaccg ccggtatctc tggtggtcac attaacccag cggtgacatt
     1021 tgggctattc ttggcacgta aagtgtcgtt acctagggcc ctattgtaca taatcgctca
     1081 gtgtttgggt gcgatttgtg gagttggttt tgtcaaagcc ttccaaagct cttactacac
     1141 ccgttacgga ggtggagcca actctctagc cgatggctac agcacaggga ccggtctagc
     1201 cgcagagatc attggtactt tcgttcttgt ctacaccgta ttctctgcca ctgaccccaa
     1261 acgtagtgcc agagactccc acgttccggt gttggcgcca cttccaatcg gatttgccgt
     1321 gttcatggta catttggcta ccattcccat taccggaacc ggaattaacc cggcaaggag
     1381 tttcggagct gccgtaatct acaacaagag caagccatgg gatgaccact ggatattttg
     1441 ggttggacca ttcattggag ctgcgatagc tgcattctac caccaattcg ttctaagagc
     1501 ttcaggttct aagtctcttg gatcattcag aagtgctgcc aacgtcggat ccatggtgag
     1561 caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat
     1621 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
     1681 cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg
     1741 ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga
     1801 catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     1861 cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt
     1921 catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa
     1981 taagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa
     2041 gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa
     2101 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa
     2161 gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga
     2221 gggccgccac tccaccggcg gcatggagga gctgtacaag taaagcggcc gcccggctgc
     2281 agatcgttca aacatttggc aataaagttt cttaagattg aatcctgttg ccggtcttgc
     2341 gatgattatc atataatttc tgttgaatta cgttaagcat gtaataatta acatgtaatg
     2401 catgacgtta tttatgagat gggtttttat gattagagtc ccgcaattat acatttaata
     2461 cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg cggtgtcatc
     2521 tatgttacta gatcggggcg cgccctcgag gaagacgtaa tgggtcccca gatttgcctt
     2581 ttcaatttca gaaagaatgc taacccacag atggttagag aggcttacgc agcagctctc
     2641 atcaagacga tctacccgag caataatctc caggaaatca aataccttcc caagaaggtt
     2701 aaagatgcag tcaaaagatt caggactaac tgcatcaaga acacagagaa agatatattt
     2761 ctcaagatca gaagtactat tccagtatgg acgattcaag gcttgcttca caaaccaagg
     2821 caagtaatag agattggagt ctctaaaaag gtagttccca ctgaatcaaa ggccatggag
     2881 tcaaagattc aaatagagga cctaacagaa ctcgccgtaa agactggcga acagttcata
     2941 cagagtctct tacgactcaa tgacaagaag aaaatcttcg tcaacatggt ggagcacgac
     3001 acacttgtct actccaaaaa tatcaaagat acagtctcag aagaccaaag ggcaattgag
     3061 acttttcaac aaagggtaat atccggaaac ctcctcggat tccattgccc agctatctgt
     3121 cactttattg tgaagatagt ggaaaaggaa ggtggctcct acaaatgcca tcattgcgat
     3181 aaaggaaagg ccatcgttga agatgcctct gccgacagtg gtcccaaaga tggaccccca
     3241 cccacgagga gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa gcaagtggat
     3301 tgatgtgata tctccactga cgtaagggat gacgcacaat cccactatcc ttcgcaagac
     3361 ccttcctcta tataaggaag ttcatttcat ttggagagaa cacgggggac tctagattga
     3421 attgattgga cgttcaatat aacaccatgg tgagtaaagg agaagaactt ttcactggag
     3481 ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg
     3541 gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg
     3601 gaaaactacc tgttccatgg ccaaccctgg tcaccaccct gacctacgta agtttctgct
     3661 tctacctttg atatatatat aataattatc attaattagt agtaatataa tatttcaaat
     3721 atttttttca aaataaaaga atgtagtata tagcaattgc ttttctgtag tttataagtg
     3781 tgtatatttt aatttataac ttttctaata tatgaccaaa atttgttgat gtgcagggcg
     3841 tgcagtgctt ctcccgttac cctgatcata tgaagcggca cgacttcttc aagagcgcca
     3901 tgcctgaggg atacgtgcag gagaggacca tcttcttcaa ggacgacggg aactacaaga
     3961 cacgtgctga agtcaagttt gagggtgaca ccctcgtcaa caggatcgag cttaagggaa
     4021 tcgatttcaa ggaggacgga aacatcctcg gccacaagtt ggaatacaac tacaactccc
     4081 acaacgtata catcatggcc gacaagcaaa agaacggcat caaagccaac ttcaagaccc
     4141 gccacaacat cgaggacggc ggcgtgcaac tcgctgatca ttatcaacaa aatactccaa
     4201 ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa tctgcccttt
     4261 cgaaagatcc caacgaaaag agagatcaca tggtccttct tgagtttgta acagctgctg
     4321 ggattacaca tggcatggat gaactataca aacctaagaa aaagcgtaag gtcgaggacc
     4381 cttaatgtca agccattctc tcgcagatga tgttcacttt gtgttttact tcctttatgc
     4441 attcacagca ataaaagaaa gaaatctcca tcgcttttgg ttttcttctc tgtcttaagt
     4501 tagtcgtttt cgtgtctaat ctattactta tcattgtaat agactcttct tctattgaga
     4561 tttgaatata aactaaaaca cattccattt tactgtgttc tcaacattca gaatgcaaac
     4621 ggactaaccg tagtgactcg gcttgagtct tcgggctaga gcggactagt tctagagcgg
     4681 ccgccaccgc ggtggagctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg
     4741 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4801 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4861 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4921 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4981 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5041 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgaag
     5101 gccttgacag gatatattgg cgggtaaact aagtcgctgt atgtgtttgt ttgagatctc
     5161 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5221 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5281 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5341 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5401 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5461 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5521 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5581 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5641 aactacggct acactagaag aacagtattt ggtatctgtg ctctgctgaa gccagttacc
     5701 ttcggaagaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5761 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5821 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5881 atgaattctt gttagacatt atttgccgac taccttggtg atctcgcctt tcacgtagtg
     5941 aacaaattct tccaactgat ctgcgcgcga ggccaagcga tcttcttgtc caagataagc
     6001 ctgcctagct tcaagtatga cgggctgata ctgggccggc aggcgctcca ttgcccagtc
     6061 ggcagcgaca tccttcggcg cgattttgcc ggttactgcg ctgtaccaaa tgcgggacaa
     6121 cgtaagcact acatttcgct catcgccagc ccagtcgggc ggcgagttcc atagcgttaa
     6181 ggtttcattt agcgcctcaa atagatcctg ttcaggaacc ggatcaaaga gttcctccgc
     6241 cgctggacct accaaggcaa cgctatgttc tcttgctttt gtcagcaaga tagccagatc
     6301 aatgtcgatc gtggctggct cgaagatacc tgcaagaatg tcattgcgct gccattctcc
     6361 aaattgcagt tcgcgcttag ctggataacg ccacggaatg atgtcgtcgt gcacaacaat
     6421 ggtgacttct acagcgcgga gaatctcgct ctctccaggg gaagccgaag tttccaaaag
     6481 gtcgttgatc aaagctcgcc gcgttgtttc atcaagcctt acggtcaccg taaccagcaa
     6541 atcaatatca ctgtgtggct tcaggccgcc atccactgcg gagccgtaca aatgtacggc
     6601 cagcaacgtc ggttcgagat ggcgctcgat gacgccaact acctctgata gttgagtcga
     6661 tacttcggcg atcaccgctt ccctcatgat gtttaactcc tgaattaagc cgcgccgcga
     6721 agcggtgtcg gcttgaatga attgttaggc gtcatcctgt gctcccgaga accagtacca
     6781 gtacatcgct gtttcgttcg agacttgagg tctagtttta tacgtgaaca ggtcaatgcc
     6841 gccgagagta aagccacatt ttgcgtacaa attgcaggca ggtacattgt tcgtttgtgc
     6901 catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga cacaacgtgg
     6961 ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc agatcacgca tcttcccgac
     7021 aacgcagacc gttccgtggc aaagcaaaag ttcaaaatca ccaactggtc cacctacaac
     7081 aaagctctca tcaaccgtgg ctccctcact ttctggctgg atgatggggc gattcaggag
     7141 atccccatcc aacagcccgc cgtcgagcgg gcttttttat ccccggaagc ctgtggatag
     7201 agggtagtta tccacgtgaa accgctaatg ccccgcaaag ccttgattca cggggctttc
     7261 cggcccgctc caaaaactat ccacgtgaaa tcgctaatca gggtacgtga aatcgctaat
     7321 cggagtacgt gaaatcgcta ataaggtcac gtgaaatcgc taatcaaaaa ggcacgtgag
     7381 aacgctaata gccctttcag atcaacagct tgcaaacacc cctcgctccg gcaagtagtt
     7441 acagcaagta gtatgttcaa ttagcttttc aattatgaat atatatatca attattggtc
     7501 gcccttggct tgtggacaat gcgctacgcg caccggctcc gcccgtggac aaccgcaagc
     7561 ggttgcccac cgtcgagcgc ctttgcccac aacccggcgg ccggccgcaa cagatcgttt
     7621 tataaatttt tttttttgaa aaagaaaaag cccgaaaggc ggcaacctct cgggcttctg
     7681 gatttccgat ccccggaatt
//
