LOCUS       sequence-391299-        7431 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants 
            (proUBQ10::PIP2A-mCherry-NOSt_proUBQ10s::GFPint-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7431)
  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 7431)
  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: JAB3064
FEATURES             Location/Qualifiers
     source          1..7431
                     /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(3234..3262)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          3402..3590
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     4046..4066
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4437..4457)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4437..4455)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4476..4492)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4476..4492)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4489..4511)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4500..4516
                     /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(4524..4530,4531..4548,4549..4554))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4569..4590
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4707..4724)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4829..4853
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4944..5532)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5024..5043)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5623..6411)
                     /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      6869..7304
                     /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 taatgtggtt gtcgacgagt cagtaataaa
     2581 cggcgtcaaa gtggttgcag ccggcacaca cgagtcgtgt ttatcaactc aaagcacaaa
     2641 tacttttcct caacctaaaa ataaggcaat tagccaaaaa caactttgcg tgtaaacaac
     2701 gctcaataca cgtgtcattt tattattagc tattgcttca ccgccttagc tttctcgtga
     2761 cctagtcgtc ctcgtctttt cttcttcttc ttctataaaa caatacccaa agagctcttc
     2821 ttcttcacaa ttcagatttc aatttctcaa aatcttaaaa actttctctc aattctctct
     2881 accgtgatca aggtaaattt ctgtgttcct tattctctca aaatcttcga ttttgttttc
     2941 gttcgatccc aatttcgtat atgttctttg gtttagattc tgttaatctt agatcgtaga
     3001 cgattttctg ggtttgatcg ttagatatca tcttaattct cgattagggt ttcatagata
     3061 tcatccgatt tgttcaaata atttgagttt tgtcgaataa ttactcttcg atttgtgatt
     3121 tctatctaga tctggtgtta gtttctagtt tgtgcgatcg aatttgtcga ttaatctgag
     3181 tttttctgat taacagcacc atggtgagta aaggagaaga acttttcact ggagttgtcc
     3241 caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc agtggagagg
     3301 gtgaaggtga tgcaacatac ggaaaactta cccttaaatt tatttgcact actggaaaac
     3361 tacctgttcc atggccaacc ctggtcacca ccctgaccta cgtaagtttc tgcttctacc
     3421 tttgatatat atataataat tatcattaat tagtagtaat ataatatttc aaatattttt
     3481 ttcaaaataa aagaatgtag tatatagcaa ttgcttttct gtagtttata agtgtgtata
     3541 ttttaattta taacttttct aatatatgac caaaatttgt tgatgtgcag ggcgtgcagt
     3601 gcttctcccg ttaccctgat catatgaagc ggcacgactt cttcaagagc gccatgcctg
     3661 agggatacgt gcaggagagg accatcttct tcaaggacga cgggaactac aagacacgtg
     3721 ctgaagtcaa gtttgagggt gacaccctcg tcaacaggat cgagcttaag ggaatcgatt
     3781 tcaaggagga cggaaacatc ctcggccaca agttggaata caactacaac tcccacaacg
     3841 tatacatcat ggccgacaag caaaagaacg gcatcaaagc caacttcaag acccgccaca
     3901 acatcgagga cggcggcgtg caactcgctg atcattatca acaaaatact ccaattggcg
     3961 atggccctgt ccttttacca gacaaccatt acctgtccac acaatctgcc ctttcgaaag
     4021 atcccaacga aaagagagat cacatggtcc ttcttgagtt tgtaacagct gctgggatta
     4081 cacatggcat ggatgaacta tacaaataat gtcaagccat tctctcgcag atgatgttca
     4141 ctttgtgttt tacttccttt atgcattcac agcaataaaa gaaagaaatc tccatcgctt
     4201 ttggttttct tctctgtctt aagttagtcg ttttcgtgtc taatctatta cttatcattg
     4261 taatagactc ttcttctatt gagatttgaa tataaactaa aacacattcc attttactgt
     4321 gttctcaaca ttcagaatgc aaacggacta accgtagtga ctcggcttga gtcttcgggc
     4381 tagagcggac tagttctaga gcggccgcca ccgcggtgga gctccagctt ttgttccctt
     4441 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4501 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4561 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4621 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4681 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4741 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4801 gataacgcag gaaagaacat gaaggccttg acaggatata ttggcgggta aactaagtcg
     4861 ctgtatgtgt ttgtttgaga tctcatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4921 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4981 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5041 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5101 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5161 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5221 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5281 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5341 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5401 tgtgctctgc tgaagccagt taccttcgga agaagagttg gtagctcttg atccggcaaa
     5461 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5521 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5581 aactcacgtt aagggatttt ggtcatgaat tcttgttaga cattatttgc cgactacctt
     5641 ggtgatctcg cctttcacgt agtgaacaaa ttcttccaac tgatctgcgc gcgaggccaa
     5701 gcgatcttct tgtccaagat aagcctgcct agcttcaagt atgacgggct gatactgggc
     5761 cggcaggcgc tccattgccc agtcggcagc gacatccttc ggcgcgattt tgccggttac
     5821 tgcgctgtac caaatgcggg acaacgtaag cactacattt cgctcatcgc cagcccagtc
     5881 gggcggcgag ttccatagcg ttaaggtttc atttagcgcc tcaaatagat cctgttcagg
     5941 aaccggatca aagagttcct ccgccgctgg acctaccaag gcaacgctat gttctcttgc
     6001 ttttgtcagc aagatagcca gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag
     6061 aatgtcattg cgctgccatt ctccaaattg cagttcgcgc ttagctggat aacgccacgg
     6121 aatgatgtcg tcgtgcacaa caatggtgac ttctacagcg cggagaatct cgctctctcc
     6181 aggggaagcc gaagtttcca aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag
     6241 ccttacggtc accgtaacca gcaaatcaat atcactgtgt ggcttcaggc cgccatccac
     6301 tgcggagccg tacaaatgta cggccagcaa cgtcggttcg agatggcgct cgatgacgcc
     6361 aactacctct gatagttgag tcgatacttc ggcgatcacc gcttccctca tgatgtttaa
     6421 ctcctgaatt aagccgcgcc gcgaagcggt gtcggcttga atgaattgtt aggcgtcatc
     6481 ctgtgctccc gagaaccagt accagtacat cgctgtttcg ttcgagactt gaggtctagt
     6541 tttatacgtg aacaggtcaa tgccgccgag agtaaagcca cattttgcgt acaaattgca
     6601 ggcaggtaca ttgttcgttt gtgccatgat gatatatttt tatcttgtgc aatgtaacat
     6661 cagagatttt gagacacaac gtggctttgt tgaataaatc gaacttttgc tgagttgaag
     6721 gatcagatca cgcatcttcc cgacaacgca gaccgttccg tggcaaagca aaagttcaaa
     6781 atcaccaact ggtccaccta caacaaagct ctcatcaacc gtggctccct cactttctgg
     6841 ctggatgatg gggcgattca ggagatcccc atccaacagc ccgccgtcga gcgggctttt
     6901 ttatccccgg aagcctgtgg atagagggta gttatccacg tgaaaccgct aatgccccgc
     6961 aaagccttga ttcacggggc tttccggccc gctccaaaaa ctatccacgt gaaatcgcta
     7021 atcagggtac gtgaaatcgc taatcggagt acgtgaaatc gctaataagg tcacgtgaaa
     7081 tcgctaatca aaaaggcacg tgagaacgct aatagccctt tcagatcaac agcttgcaaa
     7141 cacccctcgc tccggcaagt agttacagca agtagtatgt tcaattagct tttcaattat
     7201 gaatatatat atcaattatt ggtcgccctt ggcttgtgga caatgcgcta cgcgcaccgg
     7261 ctccgcccgt ggacaaccgc aagcggttgc ccaccgtcga gcgcctttgc ccacaacccg
     7321 gcggccggcc gcaacagatc gttttataaa tttttttttt tgaaaaagaa aaagcccgaa
     7381 aggcggcaac ctctcgggct tctggatttc cgatccccgg aattagactc t
//
