LOCUS       sequence-389754-        7647 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proUBQ10::PIP2A-mCherry-NOSt_proAA::GFPint-ADHter).
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: JAB3410
FEATURES             Location/Qualifiers
     source          1..7647
                     /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"
     primer_bind     complement(3450..3478)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          3618..3806
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     4262..4282
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4653..4673)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4653..4671)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4692..4708)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4692..4708)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4705..4727)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4716..4732
                     /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(4740..4746,4747..4764,4765..4770))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4785..4806
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4923..4940)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5045..5069
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5160..5748)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5240..5259)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5839..6627)
                     /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      7085..7520
                     /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 taatgggtcc ccagatttgc cttttcaatt
     2581 tcagaaagaa tgctaaccca cagatgggta gagaggctta cgcagcagct ctcatcaaga
     2641 cgatctaccc gagcaataat ctccaggaaa tcaaatacct tcccaagaag gttaaagatg
     2701 cagtcaaaag attcaggact aactgcatca agaacacaga gaaagatata tttctcaaga
     2761 tcagaagtac tattccagta tggacgattc aaggcttgct tcacaaacca aggcaagtaa
     2821 tagagattgg agtctctaaa aaggtagttc ccactgaatc aaaggccatg gagtcaaaga
     2881 ttcaaataga ggacctaaca gaactcgccg taaagactgg cgaacagttc atacagagtc
     2941 tcttacgact caatgacaag aagaaaatct tcgtcaacat ggtggagcac gacacacttg
     3001 tctactccaa aaatatcaaa gatacagtct cagaagacca aagggcaatt gagacttttc
     3061 aacaaagggt aatatccgga aacctcctcg gattccattg cccagctatc tgtcacttta
     3121 ttgtgaagat agtggaaaag gaaggtggct cctacaaatg ccatcattgc gataaaggaa
     3181 aggccatcgt tgaagatgcc tctgccgaca gtggtcccaa agatggaccc ccacccacga
     3241 ggagcatcgt ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg gattgatgtg
     3301 atatctccac tgacgtaagg gatgacgcac aatcccacta tcttctatcg atctatagat
     3361 aatctatata attctatcga tctatagata atagaacacg ggggactcta gacaccatgg
     3421 tgagtaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa ttagatggtg
     3481 atgttaatgg gcacaaattt tctgtcagtg gagagggtga aggtgatgca acatacggaa
     3541 aacttaccct taaatttatt tgcactactg gaaaactacc tgttccatgg ccaaccctgg
     3601 tcaccaccct gacctacgta agtttctgct tctacctttg atatatatat aataattatc
     3661 attaattagt agtaatataa tatttcaaat atttttttca aaataaaaga atgtagtata
     3721 tagcaattgc ttttctgtag tttataagtg tgtatatttt aatttataac ttttctaata
     3781 tatgaccaaa atttgttgat gtgcagggcg tgcagtgctt ctcccgttac cctgatcata
     3841 tgaagcggca cgacttcttc aagagcgcca tgcctgaggg atacgtgcag gagaggacca
     3901 tcttcttcaa ggacgacggg aactacaaga cacgtgctga agtcaagttt gagggtgaca
     3961 ccctcgtcaa caggatcgag cttaagggaa tcgatttcaa ggaggacgga aacatcctcg
     4021 gccacaagtt ggaatacaac tacaactccc acaacgtata catcatggcc gacaagcaaa
     4081 agaacggcat caaagccaac ttcaagaccc gccacaacat cgaggacggc ggcgtgcaac
     4141 tcgctgatca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca
     4201 accattacct gtccacacaa tctgcccttt cgaaagatcc caacgaaaag agagatcaca
     4261 tggtccttct tgagtttgta acagctgctg ggattacaca tggcatggat gaactataca
     4321 aataatgtca agccattctc tcgcagatga tgttcacttt gtgttttact tcctttatgc
     4381 attcacagca ataaaagaaa gaaatctcca tcgcttttgg ttttcttctc tgtcttaagt
     4441 tagtcgtttt cgtgtctaat ctattactta tcattgtaat agactcttct tctattgaga
     4501 tttgaatata aactaaaaca cattccattt tactgtgttc tcaacattca gaatgcaaac
     4561 ggactaaccg tagtgactcg gcttgagtct tcgggctaga gcggactagt tctagagcgg
     4621 ccgccaccgc ggtggagctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg
     4681 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4741 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4801 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4861 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4921 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4981 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgaag
     5041 gccttgacag gatatattgg cgggtaaact aagtcgctgt atgtgtttgt ttgagatctc
     5101 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5161 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5221 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5281 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5341 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5401 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5461 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5521 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5581 aactacggct acactagaag aacagtattt ggtatctgtg ctctgctgaa gccagttacc
     5641 ttcggaagaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5701 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5761 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5821 atgaattctt gttagacatt atttgccgac taccttggtg atctcgcctt tcacgtagtg
     5881 aacaaattct tccaactgat ctgcgcgcga ggccaagcga tcttcttgtc caagataagc
     5941 ctgcctagct tcaagtatga cgggctgata ctgggccggc aggcgctcca ttgcccagtc
     6001 ggcagcgaca tccttcggcg cgattttgcc ggttactgcg ctgtaccaaa tgcgggacaa
     6061 cgtaagcact acatttcgct catcgccagc ccagtcgggc ggcgagttcc atagcgttaa
     6121 ggtttcattt agcgcctcaa atagatcctg ttcaggaacc ggatcaaaga gttcctccgc
     6181 cgctggacct accaaggcaa cgctatgttc tcttgctttt gtcagcaaga tagccagatc
     6241 aatgtcgatc gtggctggct cgaagatacc tgcaagaatg tcattgcgct gccattctcc
     6301 aaattgcagt tcgcgcttag ctggataacg ccacggaatg atgtcgtcgt gcacaacaat
     6361 ggtgacttct acagcgcgga gaatctcgct ctctccaggg gaagccgaag tttccaaaag
     6421 gtcgttgatc aaagctcgcc gcgttgtttc atcaagcctt acggtcaccg taaccagcaa
     6481 atcaatatca ctgtgtggct tcaggccgcc atccactgcg gagccgtaca aatgtacggc
     6541 cagcaacgtc ggttcgagat ggcgctcgat gacgccaact acctctgata gttgagtcga
     6601 tacttcggcg atcaccgctt ccctcatgat gtttaactcc tgaattaagc cgcgccgcga
     6661 agcggtgtcg gcttgaatga attgttaggc gtcatcctgt gctcccgaga accagtacca
     6721 gtacatcgct gtttcgttcg agacttgagg tctagtttta tacgtgaaca ggtcaatgcc
     6781 gccgagagta aagccacatt ttgcgtacaa attgcaggca ggtacattgt tcgtttgtgc
     6841 catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga cacaacgtgg
     6901 ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc agatcacgca tcttcccgac
     6961 aacgcagacc gttccgtggc aaagcaaaag ttcaaaatca ccaactggtc cacctacaac
     7021 aaagctctca tcaaccgtgg ctccctcact ttctggctgg atgatggggc gattcaggag
     7081 atccccatcc aacagcccgc cgtcgagcgg gcttttttat ccccggaagc ctgtggatag
     7141 agggtagtta tccacgtgaa accgctaatg ccccgcaaag ccttgattca cggggctttc
     7201 cggcccgctc caaaaactat ccacgtgaaa tcgctaatca gggtacgtga aatcgctaat
     7261 cggagtacgt gaaatcgcta ataaggtcac gtgaaatcgc taatcaaaaa ggcacgtgag
     7321 aacgctaata gccctttcag atcaacagct tgcaaacacc cctcgctccg gcaagtagtt
     7381 acagcaagta gtatgttcaa ttagcttttc aattatgaat atatatatca attattggtc
     7441 gcccttggct tgtggacaat gcgctacgcg caccggctcc gcccgtggac aaccgcaagc
     7501 ggttgcccac cgtcgagcgc ctttgcccac aacccggcgg ccggccgcaa cagatcgttt
     7561 tataaatttt tttttttgaa aaagaaaaag cccgaaaggc ggcaacctct cgggcttctg
     7621 gatttccgat ccccggaatt agactct
//
