LOCUS       sequence-389459-        7702 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants 
            (proUBQ10::PIP2A-mCherry-NOSt_pro35S-QacRO::GFPint-NLS-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7702)
  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 7702)
  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: JAB2463
FEATURES             Location/Qualifiers
     source          1..7702
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    14..36
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     CDS             1559..2269
                     /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(1704..1722)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      2288..2540
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        3063..3408
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     3351..3371
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     complement(3488..3516)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          3656..3844
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     4300..4320
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             4361..4381
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4721..4741)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4721..4739)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4760..4776)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4760..4776)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4773..4795)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4784..4800
                     /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(4808..4814,4815..4832,4833..4838))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4853..4874
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4991..5008)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5113..5137
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5228..5816)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5308..5327)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5907..6695)
                     /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      7153..7588
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 ggaattagac tcttggcagg atatattgtg gtgtaacgtt aagctttggt tgtcgacgag
       61 tcagtaataa acggcgtcaa agtggttgca gccggcacac acgagtcgtg tttatcaact
      121 caaagcacaa atacttttcc tcaacctaaa aataaggcaa ttagccaaaa acaactttgc
      181 gtgtaaacaa cgctcaatac acgtgtcatt ttattattag ctattgcttc accgccttag
      241 ctttctcgtg acctagtcgt cctcgtcttt tcttcttctt cttctataaa acaataccca
      301 aagagctctt cttcttcaca attcagattt caatttctca aaatcttaaa aactttctct
      361 caattctctc taccgtgatc aaggtaaatt tctgtgttcc ttattctctc aaaatcttcg
      421 attttgtttt cgttcgatcc caatttcgta tatgttcttt ggtttagatt ctgttaatct
      481 tagatcgtag acgattttct gggtttgatc gttagatatc atcttaattc tcgattaggg
      541 tttcatagat atcatccgat ttgttcaaat aatttgagtt ttgtcgaata attactcttc
      601 gatttgtgat ttctatctag atctggtgtt agtttctagt ttgtgcgatc gaatttgtcg
      661 attaatctga gtttttctga ttaacagcac catggcaaag gatgtggaag ccgttcccgg
      721 agaaggattt cagacaagag actatcaaga tccgccacca gctccgttta ttgatggagc
      781 ggagctaaag aagtggtctt tctacagagc agttatcgca gagttcgtag ccactctcct
      841 cttcttatac atcaccgttt tgacagtcat cggttacaag attcagtccg atactgatgc
      901 cggtggcgta gattgcggcg gagttggaat cctcggtatc gcttgggcct ttggtggtat
      961 gatcttcatc ctcgtctact gcaccgccgg tatctctggt ggtcacatta acccagcggt
     1021 gacatttggg ctattcttgg cacgtaaagt gtcgttacct agggccctat tgtacataat
     1081 cgctcagtgt ttgggtgcga tttgtggagt tggttttgtc aaagccttcc aaagctctta
     1141 ctacacccgt tacggaggtg gagccaactc tctagccgat ggctacagca cagggaccgg
     1201 tctagccgca gagatcattg gtactttcgt tcttgtctac accgtattct ctgccactga
     1261 ccccaaacgt agtgccagag actcccacgt tccggtgttg gcgccacttc caatcggatt
     1321 tgccgtgttc atggtacatt tggctaccat tcccattacc ggaaccggaa ttaacccggc
     1381 aaggagtttc ggagctgccg taatctacaa caagagcaag ccatgggatg accactggat
     1441 attttgggtt ggaccattca ttggagctgc gatagctgca ttctaccacc aattcgttct
     1501 aagagcttca ggttctaagt ctcttggatc attcagaagt gctgccaacg tcggatccat
     1561 ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt
     1621 gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
     1681 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt
     1741 cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc
     1801 cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     1861 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg
     1921 cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat
     1981 gcagaataag accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc
     2041 cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga
     2101 ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa
     2161 catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg
     2221 cgccgagggc cgccactcca ccggcggcat ggaggagctg tacaagtaaa gcggccgccc
     2281 ggctgcagat cgttcaaaca tttggcaata aagtttctta agattgaatc ctgttgccgg
     2341 tcttgcgatg attatcatat aatttctgtt gaattacgtt aagcatgtaa taattaacat
     2401 gtaatgcatg acgttattta tgagatgggt ttttatgatt agagtcccgc aattatacat
     2461 ttaatacgcg atagaaaaca aaatatagcg cgcaaactag gataaattat cgcgcgcggt
     2521 gtcatctatg ttactagatc ggggcgcgcc ctcgaggaag acgtaatggg tccccagatt
     2581 tgccttttca atttcagaaa gaatgctaac ccacagatgg ttagagaggc ttacgcagca
     2641 gctctcatca agacgatcta cccgagcaat aatctccagg aaatcaaata ccttcccaag
     2701 aaggttaaag atgcagtcaa aagattcagg actaactgca tcaagaacac agagaaagat
     2761 atatttctca agatcagaag tactattcca gtatggacga ttcaaggctt gcttcacaaa
     2821 ccaaggcaag taatagagat tggagtctct aaaaaggtag ttcccactga atcaaaggcc
     2881 atggagtcaa agattcaaat agaggaccta acagaactcg ccgtaaagac tggcgaacag
     2941 ttcatacaga gtctcttacg actcaatgac aagaagaaaa tcttcgtcaa catggtggag
     3001 cacgacacac ttgtctactc caaaaatatc aaagatacag tctcagaaga ccaaagggca
     3061 attgagactt ttcaacaaag ggtaatatcc ggaaacctcc tcggattcca ttgcccagct
     3121 atctgtcact ttattgtgaa gatagtggaa aaggaaggtg gctcctacaa atgccatcat
     3181 tgcgataaag gaaaggccat cgttgaagat gcctctgccg acagtggtcc caaagatgga
     3241 cccccaccca cgaggagcat cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa
     3301 gtggattgat gtgatatctc cactgacgta agggatgacg cacaatccca ctatccttcg
     3361 caagaccctt cctctatata aggaagttca tttcatttgg agagaacacg ggggactcta
     3421 gattgatata gaccgtgcga tcggtctata caccatggtg agtaaaggag aagaactttt
     3481 cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc acaaattttc
     3541 tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa cttaccctta aatttatttg
     3601 cactactgga aaactacctg ttccatggcc aaccctggtc accaccctga cctacgtaag
     3661 tttctgcttc tacctttgat atatatataa taattatcat taattagtag taatataata
     3721 tttcaaatat ttttttcaaa ataaaagaat gtagtatata gcaattgctt ttctgtagtt
     3781 tataagtgtg tatattttaa tttataactt ttctaatata tgaccaaaat ttgttgatgt
     3841 gcagggcgtg cagtgcttct cccgttaccc tgatcatatg aagcggcacg acttcttcaa
     3901 gagcgccatg cctgagggat acgtgcagga gaggaccatc ttcttcaagg acgacgggaa
     3961 ctacaagaca cgtgctgaag tcaagtttga gggtgacacc ctcgtcaaca ggatcgagct
     4021 taagggaatc gatttcaagg aggacggaaa catcctcggc cacaagttgg aatacaacta
     4081 caactcccac aacgtataca tcatggccga caagcaaaag aacggcatca aagccaactt
     4141 caagacccgc cacaacatcg aggacggcgg cgtgcaactc gctgatcatt atcaacaaaa
     4201 tactccaatt ggcgatggcc ctgtcctttt accagacaac cattacctgt ccacacaatc
     4261 tgccctttcg aaagatccca acgaaaagag agatcacatg gtccttcttg agtttgtaac
     4321 agctgctggg attacacatg gcatggatga actatacaaa cctaagaaaa agcgtaaggt
     4381 cgaggaccct taatgtcaag ccattctctc gcagatgatg ttcactttgt gttttacttc
     4441 ctttatgcat tcacagcaat aaaagaaaga aatctccatc gcttttggtt ttcttctctg
     4501 tcttaagtta gtcgttttcg tgtctaatct attacttatc attgtaatag actcttcttc
     4561 tattgagatt tgaatataaa ctaaaacaca ttccatttta ctgtgttctc aacattcaga
     4621 atgcaaacgg actaaccgta gtgactcggc ttgagtcttc gggctagagc ggactagttc
     4681 tagagcggcc gccaccgcgg tggagctcca gcttttgttc cctttagtga gggttaattg
     4741 cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     4801 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     4861 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     4921 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     4981 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5041 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5101 acatgaaggc cttgacagga tatattggcg ggtaaactaa gtcgctgtat gtgtttgttt
     5161 gagatctcat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5221 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5281 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5341 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5401 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5461 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5521 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5581 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5641 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgtgct ctgctgaagc
     5701 cagttacctt cggaagaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5761 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5821 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5881 ttttggtcat gaattcttgt tagacattat ttgccgacta ccttggtgat ctcgcctttc
     5941 acgtagtgaa caaattcttc caactgatct gcgcgcgagg ccaagcgatc ttcttgtcca
     6001 agataagcct gcctagcttc aagtatgacg ggctgatact gggccggcag gcgctccatt
     6061 gcccagtcgg cagcgacatc cttcggcgcg attttgccgg ttactgcgct gtaccaaatg
     6121 cgggacaacg taagcactac atttcgctca tcgccagccc agtcgggcgg cgagttccat
     6181 agcgttaagg tttcatttag cgcctcaaat agatcctgtt caggaaccgg atcaaagagt
     6241 tcctccgccg ctggacctac caaggcaacg ctatgttctc ttgcttttgt cagcaagata
     6301 gccagatcaa tgtcgatcgt ggctggctcg aagatacctg caagaatgtc attgcgctgc
     6361 cattctccaa attgcagttc gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc
     6421 acaacaatgg tgacttctac agcgcggaga atctcgctct ctccagggga agccgaagtt
     6481 tccaaaaggt cgttgatcaa agctcgccgc gttgtttcat caagccttac ggtcaccgta
     6541 accagcaaat caatatcact gtgtggcttc aggccgccat ccactgcgga gccgtacaaa
     6601 tgtacggcca gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac ctctgatagt
     6661 tgagtcgata cttcggcgat caccgcttcc ctcatgatgt ttaactcctg aattaagccg
     6721 cgccgcgaag cggtgtcggc ttgaatgaat tgttaggcgt catcctgtgc tcccgagaac
     6781 cagtaccagt acatcgctgt ttcgttcgag acttgaggtc tagttttata cgtgaacagg
     6841 tcaatgccgc cgagagtaaa gccacatttt gcgtacaaat tgcaggcagg tacattgttc
     6901 gtttgtgcca tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca
     6961 caacgtggct ttgttgaata aatcgaactt ttgctgagtt gaaggatcag atcacgcatc
     7021 ttcccgacaa cgcagaccgt tccgtggcaa agcaaaagtt caaaatcacc aactggtcca
     7081 cctacaacaa agctctcatc aaccgtggct ccctcacttt ctggctggat gatggggcga
     7141 ttcaggagat ccccatccaa cagcccgccg tcgagcgggc ttttttatcc ccggaagcct
     7201 gtggatagag ggtagttatc cacgtgaaac cgctaatgcc ccgcaaagcc ttgattcacg
     7261 gggctttccg gcccgctcca aaaactatcc acgtgaaatc gctaatcagg gtacgtgaaa
     7321 tcgctaatcg gagtacgtga aatcgctaat aaggtcacgt gaaatcgcta atcaaaaagg
     7381 cacgtgagaa cgctaatagc cctttcagat caacagcttg caaacacccc tcgctccggc
     7441 aagtagttac agcaagtagt atgttcaatt agcttttcaa ttatgaatat atatatcaat
     7501 tattggtcgc ccttggcttg tggacaatgc gctacgcgca ccggctccgc ccgtggacaa
     7561 ccgcaagcgg ttgcccaccg tcgagcgcct ttgcccacaa cccggcggcc ggccgcaaca
     7621 gatcgtttta taaatttttt tttttgaaaa agaaaaagcc cgaaaggcgg caacctctcg
     7681 ggcttctgga tttccgatcc cc
//
