LOCUS       sequence-393623-        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-LacIO::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: JAB2652
FEATURES             Location/Qualifiers
     source          1..7699
                     /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"
     promoter        3050..3395
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     3338..3358
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     protein_bind    3416..3432
                     /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)."
     primer_bind     complement(3472..3500)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          3640..3828
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     4284..4304
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             4345..4365
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4705..4725)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4705..4723)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4744..4760)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4744..4760)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4757..4779)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4768..4784
                     /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(4792..4798,4799..4816,4817..4822))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4837..4858
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4975..4992)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5097..5121
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5212..5800)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5292..5311)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5891..6679)
                     /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      7137..7572
                     /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 cagatggtta 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 tccttcgcaa gacccttcct
     3361 ctatataagg aagttcattt catttggaga gaacacgggg gactctagat tgaaattgtg
     3421 agcggataac aattcaccat ggtgagtaaa ggagaagaac ttttcactgg agttgtccca
     3481 attcttgttg aattagatgg tgatgttaat gggcacaaat tttctgtcag tggagagggt
     3541 gaaggtgatg caacatacgg aaaacttacc cttaaattta tttgcactac tggaaaacta
     3601 cctgttccat ggccaaccct ggtcaccacc ctgacctacg taagtttctg cttctacctt
     3661 tgatatatat ataataatta tcattaatta gtagtaatat aatatttcaa atattttttt
     3721 caaaataaaa gaatgtagta tatagcaatt gcttttctgt agtttataag tgtgtatatt
     3781 ttaatttata acttttctaa tatatgacca aaatttgttg atgtgcaggg cgtgcagtgc
     3841 ttctcccgtt accctgatca tatgaagcgg cacgacttct tcaagagcgc catgcctgag
     3901 ggatacgtgc aggagaggac catcttcttc aaggacgacg ggaactacaa gacacgtgct
     3961 gaagtcaagt ttgagggtga caccctcgtc aacaggatcg agcttaaggg aatcgatttc
     4021 aaggaggacg gaaacatcct cggccacaag ttggaataca actacaactc ccacaacgta
     4081 tacatcatgg ccgacaagca aaagaacggc atcaaagcca acttcaagac ccgccacaac
     4141 atcgaggacg gcggcgtgca actcgctgat cattatcaac aaaatactcc aattggcgat
     4201 ggccctgtcc ttttaccaga caaccattac ctgtccacac aatctgccct ttcgaaagat
     4261 cccaacgaaa agagagatca catggtcctt cttgagtttg taacagctgc tgggattaca
     4321 catggcatgg atgaactata caaacctaag aaaaagcgta aggtcgagga cccttaatgt
     4381 caagccattc tctcgcagat gatgttcact ttgtgtttta cttcctttat gcattcacag
     4441 caataaaaga aagaaatctc catcgctttt ggttttcttc tctgtcttaa gttagtcgtt
     4501 ttcgtgtcta atctattact tatcattgta atagactctt cttctattga gatttgaata
     4561 taaactaaaa cacattccat tttactgtgt tctcaacatt cagaatgcaa acggactaac
     4621 cgtagtgact cggcttgagt cttcgggcta gagcggacta gttctagagc ggccgccacc
     4681 gcggtggagc tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
     4741 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4801 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4861 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4921 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4981 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5041 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatga aggccttgac
     5101 aggatatatt ggcgggtaaa ctaagtcgct gtatgtgttt gtttgagatc tcatgtgagc
     5161 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5221 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5281 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5341 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5401 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5461 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5521 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5581 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5641 ctacactaga agaacagtat ttggtatctg tgctctgctg aagccagtta ccttcggaag
     5701 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5761 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5821 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgaattc
     5881 ttgttagaca ttatttgccg actaccttgg tgatctcgcc tttcacgtag tgaacaaatt
     5941 cttccaactg atctgcgcgc gaggccaagc gatcttcttg tccaagataa gcctgcctag
     6001 cttcaagtat gacgggctga tactgggccg gcaggcgctc cattgcccag tcggcagcga
     6061 catccttcgg cgcgattttg ccggttactg cgctgtacca aatgcgggac aacgtaagca
     6121 ctacatttcg ctcatcgcca gcccagtcgg gcggcgagtt ccatagcgtt aaggtttcat
     6181 ttagcgcctc aaatagatcc tgttcaggaa ccggatcaaa gagttcctcc gccgctggac
     6241 ctaccaaggc aacgctatgt tctcttgctt ttgtcagcaa gatagccaga tcaatgtcga
     6301 tcgtggctgg ctcgaagata cctgcaagaa tgtcattgcg ctgccattct ccaaattgca
     6361 gttcgcgctt agctggataa cgccacggaa tgatgtcgtc gtgcacaaca atggtgactt
     6421 ctacagcgcg gagaatctcg ctctctccag gggaagccga agtttccaaa aggtcgttga
     6481 tcaaagctcg ccgcgttgtt tcatcaagcc ttacggtcac cgtaaccagc aaatcaatat
     6541 cactgtgtgg cttcaggccg ccatccactg cggagccgta caaatgtacg gccagcaacg
     6601 tcggttcgag atggcgctcg atgacgccaa ctacctctga tagttgagtc gatacttcgg
     6661 cgatcaccgc ttccctcatg atgtttaact cctgaattaa gccgcgccgc gaagcggtgt
     6721 cggcttgaat gaattgttag gcgtcatcct gtgctcccga gaaccagtac cagtacatcg
     6781 ctgtttcgtt cgagacttga ggtctagttt tatacgtgaa caggtcaatg ccgccgagag
     6841 taaagccaca ttttgcgtac aaattgcagg caggtacatt gttcgtttgt gccatgatga
     6901 tatattttta tcttgtgcaa tgtaacatca gagattttga gacacaacgt ggctttgttg
     6961 aataaatcga acttttgctg agttgaagga tcagatcacg catcttcccg acaacgcaga
     7021 ccgttccgtg gcaaagcaaa agttcaaaat caccaactgg tccacctaca acaaagctct
     7081 catcaaccgt ggctccctca ctttctggct ggatgatggg gcgattcagg agatccccat
     7141 ccaacagccc gccgtcgagc gggctttttt atccccggaa gcctgtggat agagggtagt
     7201 tatccacgtg aaaccgctaa tgccccgcaa agccttgatt cacggggctt tccggcccgc
     7261 tccaaaaact atccacgtga aatcgctaat cagggtacgt gaaatcgcta atcggagtac
     7321 gtgaaatcgc taataaggtc acgtgaaatc gctaatcaaa aaggcacgtg agaacgctaa
     7381 tagccctttc agatcaacag cttgcaaaca cccctcgctc cggcaagtag ttacagcaag
     7441 tagtatgttc aattagcttt tcaattatga atatatatat caattattgg tcgcccttgg
     7501 cttgtggaca atgcgctacg cgcaccggct ccgcccgtgg acaaccgcaa gcggttgccc
     7561 accgtcgagc gcctttgccc acaacccggc ggccggccgc aacagatcgt tttataaatt
     7621 tttttttttg aaaaagaaaa agcccgaaag gcggcaacct ctcgggcttc tggatttccg
     7681 atccccggaa ttagactct
//
