LOCUS       sequence-389486-        7697 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants 
            (proUBQ10::PIP2A-mCherry-NOSt_pro35S-TetRO::GFPint-NLS-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7697)
  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 7697)
  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: JAB2515
FEATURES             Location/Qualifiers
     source          1..7697
                     /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"
     protein_bind    3426..3444
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     primer_bind     complement(3482..3510)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          3650..3838
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     4294..4314
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             4355..4375
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4715..4735)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4715..4733)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4754..4770)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4754..4770)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4767..4789)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4778..4794
                     /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(4802..4808,4809..4826,4827..4832))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4847..4868
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4985..5002)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5107..5131
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5222..5810)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5302..5321)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5901..6689)
                     /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      7147..7582
                     /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 attgaactct atcattgata gagtcaccat ggtgagtaaa ggagaagaac ttttcactgg
     3481 agttgtccca attcttgttg aattagatgg tgatgttaat gggcacaaat tttctgtcag
     3541 tggagagggt gaaggtgatg caacatacgg aaaacttacc cttaaattta tttgcactac
     3601 tggaaaacta cctgttccat ggccaaccct ggtcaccacc ctgacctacg taagtttctg
     3661 cttctacctt tgatatatat ataataatta tcattaatta gtagtaatat aatatttcaa
     3721 atattttttt caaaataaaa gaatgtagta tatagcaatt gcttttctgt agtttataag
     3781 tgtgtatatt ttaatttata acttttctaa tatatgacca aaatttgttg atgtgcaggg
     3841 cgtgcagtgc ttctcccgtt accctgatca tatgaagcgg cacgacttct tcaagagcgc
     3901 catgcctgag ggatacgtgc aggagaggac catcttcttc aaggacgacg ggaactacaa
     3961 gacacgtgct gaagtcaagt ttgagggtga caccctcgtc aacaggatcg agcttaaggg
     4021 aatcgatttc aaggaggacg gaaacatcct cggccacaag ttggaataca actacaactc
     4081 ccacaacgta tacatcatgg ccgacaagca aaagaacggc atcaaagcca acttcaagac
     4141 ccgccacaac atcgaggacg gcggcgtgca actcgctgat cattatcaac aaaatactcc
     4201 aattggcgat ggccctgtcc ttttaccaga caaccattac ctgtccacac aatctgccct
     4261 ttcgaaagat cccaacgaaa agagagatca catggtcctt cttgagtttg taacagctgc
     4321 tgggattaca catggcatgg atgaactata caaacctaag aaaaagcgta aggtcgagga
     4381 cccttaatgt caagccattc tctcgcagat gatgttcact ttgtgtttta cttcctttat
     4441 gcattcacag caataaaaga aagaaatctc catcgctttt ggttttcttc tctgtcttaa
     4501 gttagtcgtt ttcgtgtcta atctattact tatcattgta atagactctt cttctattga
     4561 gatttgaata taaactaaaa cacattccat tttactgtgt tctcaacatt cagaatgcaa
     4621 acggactaac cgtagtgact cggcttgagt cttcgggcta gagcggacta gttctagagc
     4681 ggccgccacc gcggtggagc tccagctttt gttcccttta gtgagggtta attgcgcgct
     4741 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     4801 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     4861 tcacattaat tgcgttgcgc tcactgccag ctttccagtc gggaaacctg tcgtgccagc
     4921 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     4981 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5041 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatga
     5101 aggccttgac aggatatatt ggcgggtaaa ctaagtcgct gtatgtgttt gtttgagatc
     5161 tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5221 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5281 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5341 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5401 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5461 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5521 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5581 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5641 ctaactacgg ctacactaga agaacagtat ttggtatctg tgctctgctg aagccagtta
     5701 ccttcggaag aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5761 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5821 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5881 tcatgaattc ttgttagaca ttatttgccg actaccttgg tgatctcgcc tttcacgtag
     5941 tgaacaaatt cttccaactg atctgcgcgc gaggccaagc gatcttcttg tccaagataa
     6001 gcctgcctag cttcaagtat gacgggctga tactgggccg gcaggcgctc cattgcccag
     6061 tcggcagcga catccttcgg cgcgattttg ccggttactg cgctgtacca aatgcgggac
     6121 aacgtaagca ctacatttcg ctcatcgcca gcccagtcgg gcggcgagtt ccatagcgtt
     6181 aaggtttcat ttagcgcctc aaatagatcc tgttcaggaa ccggatcaaa gagttcctcc
     6241 gccgctggac ctaccaaggc aacgctatgt tctcttgctt ttgtcagcaa gatagccaga
     6301 tcaatgtcga tcgtggctgg ctcgaagata cctgcaagaa tgtcattgcg ctgccattct
     6361 ccaaattgca gttcgcgctt agctggataa cgccacggaa tgatgtcgtc gtgcacaaca
     6421 atggtgactt ctacagcgcg gagaatctcg ctctctccag gggaagccga agtttccaaa
     6481 aggtcgttga tcaaagctcg ccgcgttgtt tcatcaagcc ttacggtcac cgtaaccagc
     6541 aaatcaatat cactgtgtgg cttcaggccg ccatccactg cggagccgta caaatgtacg
     6601 gccagcaacg tcggttcgag atggcgctcg atgacgccaa ctacctctga tagttgagtc
     6661 gatacttcgg cgatcaccgc ttccctcatg atgtttaact cctgaattaa gccgcgccgc
     6721 gaagcggtgt cggcttgaat gaattgttag gcgtcatcct gtgctcccga gaaccagtac
     6781 cagtacatcg ctgtttcgtt cgagacttga ggtctagttt tatacgtgaa caggtcaatg
     6841 ccgccgagag taaagccaca ttttgcgtac aaattgcagg caggtacatt gttcgtttgt
     6901 gccatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga gacacaacgt
     6961 ggctttgttg aataaatcga acttttgctg agttgaagga tcagatcacg catcttcccg
     7021 acaacgcaga ccgttccgtg gcaaagcaaa agttcaaaat caccaactgg tccacctaca
     7081 acaaagctct catcaaccgt ggctccctca ctttctggct ggatgatggg gcgattcagg
     7141 agatccccat ccaacagccc gccgtcgagc gggctttttt atccccggaa gcctgtggat
     7201 agagggtagt tatccacgtg aaaccgctaa tgccccgcaa agccttgatt cacggggctt
     7261 tccggcccgc tccaaaaact atccacgtga aatcgctaat cagggtacgt gaaatcgcta
     7321 atcggagtac gtgaaatcgc taataaggtc acgtgaaatc gctaatcaaa aaggcacgtg
     7381 agaacgctaa tagccctttc agatcaacag cttgcaaaca cccctcgctc cggcaagtag
     7441 ttacagcaag tagtatgttc aattagcttt tcaattatga atatatatat caattattgg
     7501 tcgcccttgg cttgtggaca atgcgctacg cgcaccggct ccgcccgtgg acaaccgcaa
     7561 gcggttgccc accgtcgagc gcctttgccc acaacccggc ggccggccgc aacagatcgt
     7621 tttataaatt tttttttttg aaaaagaaaa agcccgaaag gcggcaacct ctcgggcttc
     7681 tggatttccg atccccg
//
