LOCUS       sequence-389441-        7699 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants 
            (proUBQ10::PIP2A-mCherry-NOSt_pro35S-AmtRO::GFPint-NLS-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7699)
  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 7699)
  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: JAB2275
FEATURES             Location/Qualifiers
     source          1..7699
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    13..35
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     CDS             1558..2268
                     /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(1703..1721)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      2287..2539
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        3062..3407
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     3350..3370
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     complement(3484..3512)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          3652..3840
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     4296..4316
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             4357..4377
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4717..4737)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4717..4735)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4756..4772)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4756..4772)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4769..4791)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4780..4796
                     /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(4804..4810,4811..4828,4829..4834))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4849..4870
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4987..5004)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5109..5133
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5224..5812)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5304..5323)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5903..6691)
                     /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      7149..7584
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 gaattagact cttggcagga tatattgtgg tgtaacgtta agctttggtt gtcgacgagt
       61 cagtaataaa cggcgtcaaa gtggttgcag ccggcacaca cgagtcgtgt ttatcaactc
      121 aaagcacaaa tacttttcct caacctaaaa ataaggcaat tagccaaaaa caactttgcg
      181 tgtaaacaac gctcaataca cgtgtcattt tattattagc tattgcttca ccgccttagc
      241 tttctcgtga cctagtcgtc ctcgtctttt cttcttcttc ttctataaaa caatacccaa
      301 agagctcttc ttcttcacaa ttcagatttc aatttctcaa aatcttaaaa actttctctc
      361 aattctctct accgtgatca aggtaaattt ctgtgttcct tattctctca aaatcttcga
      421 ttttgttttc gttcgatccc aatttcgtat atgttctttg gtttagattc tgttaatctt
      481 agatcgtaga cgattttctg ggtttgatcg ttagatatca tcttaattct cgattagggt
      541 ttcatagata tcatccgatt tgttcaaata atttgagttt tgtcgaataa ttactcttcg
      601 atttgtgatt tctatctaga tctggtgtta gtttctagtt tgtgcgatcg aatttgtcga
      661 ttaatctgag tttttctgat taacagcacc atggcaaagg atgtggaagc cgttcccgga
      721 gaaggatttc agacaagaga ctatcaagat ccgccaccag ctccgtttat tgatggagcg
      781 gagctaaaga agtggtcttt ctacagagca gttatcgcag agttcgtagc cactctcctc
      841 ttcttataca tcaccgtttt gacagtcatc ggttacaaga ttcagtccga tactgatgcc
      901 ggtggcgtag attgcggcgg agttggaatc ctcggtatcg cttgggcctt tggtggtatg
      961 atcttcatcc tcgtctactg caccgccggt atctctggtg gtcacattaa cccagcggtg
     1021 acatttgggc tattcttggc acgtaaagtg tcgttaccta gggccctatt gtacataatc
     1081 gctcagtgtt tgggtgcgat ttgtggagtt ggttttgtca aagccttcca aagctcttac
     1141 tacacccgtt acggaggtgg agccaactct ctagccgatg gctacagcac agggaccggt
     1201 ctagccgcag agatcattgg tactttcgtt cttgtctaca ccgtattctc tgccactgac
     1261 cccaaacgta gtgccagaga ctcccacgtt ccggtgttgg cgccacttcc aatcggattt
     1321 gccgtgttca tggtacattt ggctaccatt cccattaccg gaaccggaat taacccggca
     1381 aggagtttcg gagctgccgt aatctacaac aagagcaagc catgggatga ccactggata
     1441 ttttgggttg gaccattcat tggagctgcg atagctgcat tctaccacca attcgttcta
     1501 agagcttcag gttctaagtc tcttggatca ttcagaagtg ctgccaacgt cggatccatg
     1561 gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg
     1621 cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
     1681 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc
     1741 gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc
     1801 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
     1861 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
     1921 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
     1981 cagaataaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc
     2041 ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag
     2101 gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac
     2161 atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc
     2221 gccgagggcc gccactccac cggcggcatg gaggagctgt acaagtaaag cggccgcccg
     2281 gctgcagatc gttcaaacat ttggcaataa agtttcttaa gattgaatcc tgttgccggt
     2341 cttgcgatga ttatcatata atttctgttg aattacgtta agcatgtaat aattaacatg
     2401 taatgcatga cgttatttat gagatgggtt tttatgatta gagtcccgca attatacatt
     2461 taatacgcga tagaaaacaa aatatagcgc gcaaactagg ataaattatc gcgcgcggtg
     2521 tcatctatgt tactagatcg gggcgcgccc tcgaggaaga cgtaatgggt ccccagattt
     2581 gccttttcaa tttcagaaag aatgctaacc cacagatggt tagagaggct tacgcagcag
     2641 ctctcatcaa gacgatctac ccgagcaata atctccagga aatcaaatac cttcccaaga
     2701 aggttaaaga tgcagtcaaa agattcagga ctaactgcat caagaacaca gagaaagata
     2761 tatttctcaa gatcagaagt actattccag tatggacgat tcaaggcttg cttcacaaac
     2821 caaggcaagt aatagagatt ggagtctcta aaaaggtagt tcccactgaa tcaaaggcca
     2881 tggagtcaaa gattcaaata gaggacctaa cagaactcgc cgtaaagact ggcgaacagt
     2941 tcatacagag tctcttacga ctcaatgaca agaagaaaat cttcgtcaac atggtggagc
     3001 acgacacact tgtctactcc aaaaatatca aagatacagt ctcagaagac caaagggcaa
     3061 ttgagacttt tcaacaaagg gtaatatccg gaaacctcct cggattccat tgcccagcta
     3121 tctgtcactt tattgtgaag atagtggaaa aggaaggtgg ctcctacaaa tgccatcatt
     3181 gcgataaagg aaaggccatc gttgaagatg cctctgccga cagtggtccc aaagatggac
     3241 ccccacccac gaggagcatc gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag
     3301 tggattgatg tgatatctcc actgacgtaa gggatgacgc acaatcccac tatccttcgc
     3361 aagacccttc ctctatataa ggaagttcat ttcatttgga gagaacacgg gggactctag
     3421 attgattcta tcgatctata gataatcacc atggtgagta aaggagaaga acttttcact
     3481 ggagttgtcc caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc
     3541 agtggagagg gtgaaggtga tgcaacatac ggaaaactta cccttaaatt tatttgcact
     3601 actggaaaac tacctgttcc atggccaacc ctggtcacca ccctgaccta cgtaagtttc
     3661 tgcttctacc tttgatatat atataataat tatcattaat tagtagtaat ataatatttc
     3721 aaatattttt ttcaaaataa aagaatgtag tatatagcaa ttgcttttct gtagtttata
     3781 agtgtgtata ttttaattta taacttttct aatatatgac caaaatttgt tgatgtgcag
     3841 ggcgtgcagt gcttctcccg ttaccctgat catatgaagc ggcacgactt cttcaagagc
     3901 gccatgcctg agggatacgt gcaggagagg accatcttct tcaaggacga cgggaactac
     3961 aagacacgtg ctgaagtcaa gtttgagggt gacaccctcg tcaacaggat cgagcttaag
     4021 ggaatcgatt tcaaggagga cggaaacatc ctcggccaca agttggaata caactacaac
     4081 tcccacaacg tatacatcat ggccgacaag caaaagaacg gcatcaaagc caacttcaag
     4141 acccgccaca acatcgagga cggcggcgtg caactcgctg atcattatca acaaaatact
     4201 ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccac acaatctgcc
     4261 ctttcgaaag atcccaacga aaagagagat cacatggtcc ttcttgagtt tgtaacagct
     4321 gctgggatta cacatggcat ggatgaacta tacaaaccta agaaaaagcg taaggtcgag
     4381 gacccttaat gtcaagccat tctctcgcag atgatgttca ctttgtgttt tacttccttt
     4441 atgcattcac agcaataaaa gaaagaaatc tccatcgctt ttggttttct tctctgtctt
     4501 aagttagtcg ttttcgtgtc taatctatta cttatcattg taatagactc ttcttctatt
     4561 gagatttgaa tataaactaa aacacattcc attttactgt gttctcaaca ttcagaatgc
     4621 aaacggacta accgtagtga ctcggcttga gtcttcgggc tagagcggac tagttctaga
     4681 gcggccgcca ccgcggtgga gctccagctt ttgttccctt tagtgagggt taattgcgcg
     4741 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4801 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4861 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4921 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4981 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5041 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5101 gaaggccttg acaggatata ttggcgggta aactaagtcg ctgtatgtgt ttgtttgaga
     5161 tctcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5221 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5281 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5341 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5401 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5461 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5521 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5581 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5641 gcctaactac ggctacacta gaagaacagt atttggtatc tgtgctctgc tgaagccagt
     5701 taccttcgga agaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5761 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5821 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5881 ggtcatgaat tcttgttaga cattatttgc cgactacctt ggtgatctcg cctttcacgt
     5941 agtgaacaaa ttcttccaac tgatctgcgc gcgaggccaa gcgatcttct tgtccaagat
     6001 aagcctgcct agcttcaagt atgacgggct gatactgggc cggcaggcgc tccattgccc
     6061 agtcggcagc gacatccttc ggcgcgattt tgccggttac tgcgctgtac caaatgcggg
     6121 acaacgtaag cactacattt cgctcatcgc cagcccagtc gggcggcgag ttccatagcg
     6181 ttaaggtttc atttagcgcc tcaaatagat cctgttcagg aaccggatca aagagttcct
     6241 ccgccgctgg acctaccaag gcaacgctat gttctcttgc ttttgtcagc aagatagcca
     6301 gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag aatgtcattg cgctgccatt
     6361 ctccaaattg cagttcgcgc ttagctggat aacgccacgg aatgatgtcg tcgtgcacaa
     6421 caatggtgac ttctacagcg cggagaatct cgctctctcc aggggaagcc gaagtttcca
     6481 aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag ccttacggtc accgtaacca
     6541 gcaaatcaat atcactgtgt ggcttcaggc cgccatccac tgcggagccg tacaaatgta
     6601 cggccagcaa cgtcggttcg agatggcgct cgatgacgcc aactacctct gatagttgag
     6661 tcgatacttc ggcgatcacc gcttccctca tgatgtttaa ctcctgaatt aagccgcgcc
     6721 gcgaagcggt gtcggcttga atgaattgtt aggcgtcatc ctgtgctccc gagaaccagt
     6781 accagtacat cgctgtttcg ttcgagactt gaggtctagt tttatacgtg aacaggtcaa
     6841 tgccgccgag agtaaagcca cattttgcgt acaaattgca ggcaggtaca ttgttcgttt
     6901 gtgccatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt gagacacaac
     6961 gtggctttgt tgaataaatc gaacttttgc tgagttgaag gatcagatca cgcatcttcc
     7021 cgacaacgca gaccgttccg tggcaaagca aaagttcaaa atcaccaact ggtccaccta
     7081 caacaaagct ctcatcaacc gtggctccct cactttctgg ctggatgatg gggcgattca
     7141 ggagatcccc atccaacagc ccgccgtcga gcgggctttt ttatccccgg aagcctgtgg
     7201 atagagggta gttatccacg tgaaaccgct aatgccccgc aaagccttga ttcacggggc
     7261 tttccggccc gctccaaaaa ctatccacgt gaaatcgcta atcagggtac gtgaaatcgc
     7321 taatcggagt acgtgaaatc gctaataagg tcacgtgaaa tcgctaatca aaaaggcacg
     7381 tgagaacgct aatagccctt tcagatcaac agcttgcaaa cacccctcgc tccggcaagt
     7441 agttacagca agtagtatgt tcaattagct tttcaattat gaatatatat atcaattatt
     7501 ggtcgccctt ggcttgtgga caatgcgcta cgcgcaccgg ctccgcccgt ggacaaccgc
     7561 aagcggttgc ccaccgtcga gcgcctttgc ccacaacccg gcggccggcc gcaacagatc
     7621 gttttataaa tttttttttt tgaaaaagaa aaagcccgaa aggcggcaac ctctcgggct
     7681 tctggatttc cgatccccg
//
