LOCUS       sequence-389759-        7676 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-NLS_ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7676)
  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 7676)
  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: JAB2938
FEATURES             Location/Qualifiers
     source          1..7676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    42..64
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     CDS             1587..2297
                     /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(1732..1750)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      2316..2568
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     3379..3399
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     complement(3490..3518)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          3658..3846
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     4302..4322
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             4363..4383
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4723..4743)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4723..4741)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4762..4778)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4762..4778)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4775..4797)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4786..4802
                     /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(4810..4816,4817..4834,4835..4840))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4855..4876
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4993..5010)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5115..5139
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5230..5818)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5310..5329)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5909..6697)
                     /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      7155..7590
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 ctctcgggct tctggatttc cgatccccgg aattagactc ttggcaggat atattgtggt
       61 gtaacgttaa gctttggttg tcgacgagtc agtaataaac ggcgtcaaag tggttgcagc
      121 cggcacacac gagtcgtgtt tatcaactca aagcacaaat acttttcctc aacctaaaaa
      181 taaggcaatt agccaaaaac aactttgcgt gtaaacaacg ctcaatacac gtgtcatttt
      241 attattagct attgcttcac cgccttagct ttctcgtgac ctagtcgtcc tcgtcttttc
      301 ttcttcttct tctataaaac aatacccaaa gagctcttct tcttcacaat tcagatttca
      361 atttctcaaa atcttaaaaa ctttctctca attctctcta ccgtgatcaa ggtaaatttc
      421 tgtgttcctt attctctcaa aatcttcgat tttgttttcg ttcgatccca atttcgtata
      481 tgttctttgg tttagattct gttaatctta gatcgtagac gattttctgg gtttgatcgt
      541 tagatatcat cttaattctc gattagggtt tcatagatat catccgattt gttcaaataa
      601 tttgagtttt gtcgaataat tactcttcga tttgtgattt ctatctagat ctggtgttag
      661 tttctagttt gtgcgatcga atttgtcgat taatctgagt ttttctgatt aacagcacca
      721 tggcaaagga tgtggaagcc gttcccggag aaggatttca gacaagagac tatcaagatc
      781 cgccaccagc tccgtttatt gatggagcgg agctaaagaa gtggtctttc tacagagcag
      841 ttatcgcaga gttcgtagcc actctcctct tcttatacat caccgttttg acagtcatcg
      901 gttacaagat tcagtccgat actgatgccg gtggcgtaga ttgcggcgga gttggaatcc
      961 tcggtatcgc ttgggccttt ggtggtatga tcttcatcct cgtctactgc accgccggta
     1021 tctctggtgg tcacattaac ccagcggtga catttgggct attcttggca cgtaaagtgt
     1081 cgttacctag ggccctattg tacataatcg ctcagtgttt gggtgcgatt tgtggagttg
     1141 gttttgtcaa agccttccaa agctcttact acacccgtta cggaggtgga gccaactctc
     1201 tagccgatgg ctacagcaca gggaccggtc tagccgcaga gatcattggt actttcgttc
     1261 ttgtctacac cgtattctct gccactgacc ccaaacgtag tgccagagac tcccacgttc
     1321 cggtgttggc gccacttcca atcggatttg ccgtgttcat ggtacatttg gctaccattc
     1381 ccattaccgg aaccggaatt aacccggcaa ggagtttcgg agctgccgta atctacaaca
     1441 agagcaagcc atgggatgac cactggatat tttgggttgg accattcatt ggagctgcga
     1501 tagctgcatt ctaccaccaa ttcgttctaa gagcttcagg ttctaagtct cttggatcat
     1561 tcagaagtgc tgccaacgtc ggatccatgg tgagcaaggg cgaggaggat aacatggcca
     1621 tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt
     1681 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
     1741 aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt
     1801 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct
     1861 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
     1921 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
     1981 ccaacttccc ctccgacggc cccgtaatgc agaataagac catgggctgg gaggcctcct
     2041 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc
     2101 tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg
     2161 tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg
     2221 actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg
     2281 aggagctgta caagtaaagc ggccgcccgg ctgcagatcg ttcaaacatt tggcaataaa
     2341 gtttcttaag attgaatcct gttgccggtc ttgcgatgat tatcatataa tttctgttga
     2401 attacgttaa gcatgtaata attaacatgt aatgcatgac gttatttatg agatgggttt
     2461 ttatgattag agtcccgcaa ttatacattt aatacgcgat agaaaacaaa atatagcgcg
     2521 caaactagga taaattatcg cgcgcggtgt catctatgtt actagatcgg ggcgcgccct
     2581 cgaggaagac gtaatgggtc cccagatttg ccttttcaat ttcagaaaga atgctaaccc
     2641 acagatggtt agagaggctt acgcagcagc tctcatcaag acgatctacc cgagcaataa
     2701 tctccaggaa atcaaatacc ttcccaagaa ggttaaagat gcagtcaaaa gattcaggac
     2761 taactgcatc aagaacacag agaaagatat atttctcaag atcagaagta ctattccagt
     2821 atggacgatt caaggcttgc ttcacaaacc aaggcaagta atagagattg gagtctctaa
     2881 aaaggtagtt cccactgaat caaaggccat ggagtcaaag attcaaatag aggacctaac
     2941 agaactcgcc gtaaagactg gcgaacagtt catacagagt ctcttacgac tcaatgacaa
     3001 gaagaaaatc ttcgtcaaca tggtggagca cgacacactt gtctactcca aaaatatcaa
     3061 agatacagtc tcagaagacc aaagggcaat tgagactttt caacaaaggg taatatccgg
     3121 aaacctcctc ggattccatt gcccagctat ctgtcacttt attgtgaaga tagtggaaaa
     3181 ggaaggtggc tcctacaaat gccatcattg cgataaagga aaggccatcg ttgaagatgc
     3241 ctctgccgac agtggtccca aagatggacc cccacccacg aggagcatcg tggaaaaaga
     3301 agacgttcca accacgtctt caaagcaagt ggattgatgt gatatctcca ctgacgtaag
     3361 ggatgacgca caatcccact atccttcgca agacccttcc tctatataaa ctctatcact
     3421 gatagagtgg actctatcac tgatagagta gacaccatgg tgagtaaagg agaagaactt
     3481 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
     3541 tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt
     3601 tgcactactg gaaaactacc tgttccatgg ccaaccctgg tcaccaccct gacctacgta
     3661 agtttctgct tctacctttg atatatatat aataattatc attaattagt agtaatataa
     3721 tatttcaaat atttttttca aaataaaaga atgtagtata tagcaattgc ttttctgtag
     3781 tttataagtg tgtatatttt aatttataac ttttctaata tatgaccaaa atttgttgat
     3841 gtgcagggcg tgcagtgctt ctcccgttac cctgatcata tgaagcggca cgacttcttc
     3901 aagagcgcca tgcctgaggg atacgtgcag gagaggacca tcttcttcaa ggacgacggg
     3961 aactacaaga cacgtgctga agtcaagttt gagggtgaca ccctcgtcaa caggatcgag
     4021 cttaagggaa tcgatttcaa ggaggacgga aacatcctcg gccacaagtt ggaatacaac
     4081 tacaactccc acaacgtata catcatggcc gacaagcaaa agaacggcat caaagccaac
     4141 ttcaagaccc gccacaacat cgaggacggc ggcgtgcaac tcgctgatca ttatcaacaa
     4201 aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa
     4261 tctgcccttt cgaaagatcc caacgaaaag agagatcaca tggtccttct tgagtttgta
     4321 acagctgctg ggattacaca tggcatggat gaactataca aacctaagaa aaagcgtaag
     4381 gtcgaggacc cttaatgtca agccattctc tcgcagatga tgttcacttt gtgttttact
     4441 tcctttatgc attcacagca ataaaagaaa gaaatctcca tcgcttttgg ttttcttctc
     4501 tgtcttaagt tagtcgtttt cgtgtctaat ctattactta tcattgtaat agactcttct
     4561 tctattgaga tttgaatata aactaaaaca cattccattt tactgtgttc tcaacattca
     4621 gaatgcaaac ggactaaccg tagtgactcg gcttgagtct tcgggctaga gcggactagt
     4681 tctagagcgg ccgccaccgc ggtggagctc cagcttttgt tccctttagt gagggttaat
     4741 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4801 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4861 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4921 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4981 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5041 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5101 gaacatgaag gccttgacag gatatattgg cgggtaaact aagtcgctgt atgtgtttgt
     5161 ttgagatctc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5221 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5281 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5341 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5401 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5461 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5521 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5581 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5641 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgtg ctctgctgaa
     5701 gccagttacc ttcggaagaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5761 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5821 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5881 gattttggtc atgaattctt gttagacatt atttgccgac taccttggtg atctcgcctt
     5941 tcacgtagtg aacaaattct tccaactgat ctgcgcgcga ggccaagcga tcttcttgtc
     6001 caagataagc ctgcctagct tcaagtatga cgggctgata ctgggccggc aggcgctcca
     6061 ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc ggttactgcg ctgtaccaaa
     6121 tgcgggacaa cgtaagcact acatttcgct catcgccagc ccagtcgggc ggcgagttcc
     6181 atagcgttaa ggtttcattt agcgcctcaa atagatcctg ttcaggaacc ggatcaaaga
     6241 gttcctccgc cgctggacct accaaggcaa cgctatgttc tcttgctttt gtcagcaaga
     6301 tagccagatc aatgtcgatc gtggctggct cgaagatacc tgcaagaatg tcattgcgct
     6361 gccattctcc aaattgcagt tcgcgcttag ctggataacg ccacggaatg atgtcgtcgt
     6421 gcacaacaat ggtgacttct acagcgcgga gaatctcgct ctctccaggg gaagccgaag
     6481 tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc atcaagcctt acggtcaccg
     6541 taaccagcaa atcaatatca ctgtgtggct tcaggccgcc atccactgcg gagccgtaca
     6601 aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat gacgccaact acctctgata
     6661 gttgagtcga tacttcggcg atcaccgctt ccctcatgat gtttaactcc tgaattaagc
     6721 cgcgccgcga agcggtgtcg gcttgaatga attgttaggc gtcatcctgt gctcccgaga
     6781 accagtacca gtacatcgct gtttcgttcg agacttgagg tctagtttta tacgtgaaca
     6841 ggtcaatgcc gccgagagta aagccacatt ttgcgtacaa attgcaggca ggtacattgt
     6901 tcgtttgtgc catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga
     6961 cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc agatcacgca
     7021 tcttcccgac aacgcagacc gttccgtggc aaagcaaaag ttcaaaatca ccaactggtc
     7081 cacctacaac aaagctctca tcaaccgtgg ctccctcact ttctggctgg atgatggggc
     7141 gattcaggag atccccatcc aacagcccgc cgtcgagcgg gcttttttat ccccggaagc
     7201 ctgtggatag agggtagtta tccacgtgaa accgctaatg ccccgcaaag ccttgattca
     7261 cggggctttc cggcccgctc caaaaactat ccacgtgaaa tcgctaatca gggtacgtga
     7321 aatcgctaat cggagtacgt gaaatcgcta ataaggtcac gtgaaatcgc taatcaaaaa
     7381 ggcacgtgag aacgctaata gccctttcag atcaacagct tgcaaacacc cctcgctccg
     7441 gcaagtagtt acagcaagta gtatgttcaa ttagcttttc aattatgaat atatatatca
     7501 attattggtc gcccttggct tgtggacaat gcgctacgcg caccggctcc gcccgtggac
     7561 aaccgcaagc ggttgcccac cgtcgagcgc ctttgcccac aacccggcgg ccggccgcaa
     7621 cagatcgttt tataaatttt tttttttgaa aaagaaaaag cccgaaaggc ggcaac
//
