LOCUS       sequence-387075-        7047 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants 
            (proUBQ10::PIP2A-mCherry-NOSt_6XTetRO-min35S::GFPint-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7047)
  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 7047)
  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: JAB2193
FEATURES             Location/Qualifiers
     source          1..7047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    16..38
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     CDS             1561..2271
                     /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(1706..1724)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      2290..2542
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    2597..2615
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    2618..2636
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    2639..2657
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    2660..2678
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    2681..2699
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    2702..2720
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     promoter        2765..2811
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     primer_bind     complement(2865..2893)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          3033..3221
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     3677..3697
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4068..4088)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4068..4086)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4107..4123)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4107..4123)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4120..4142)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4131..4147
                     /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(4155..4161,4162..4179,4180..4185))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4200..4221
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4338..4355)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4460..4484
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4575..5163)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4655..4674)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5254..6042)
                     /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      6500..6935
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 ccggaattag actcttggca ggatatattg tggtgtaacg ttaagctttg gttgtcgacg
       61 agtcagtaat aaacggcgtc aaagtggttg cagccggcac acacgagtcg tgtttatcaa
      121 ctcaaagcac aaatactttt cctcaaccta aaaataaggc aattagccaa aaacaacttt
      181 gcgtgtaaac aacgctcaat acacgtgtca ttttattatt agctattgct tcaccgcctt
      241 agctttctcg tgacctagtc gtcctcgtct tttcttcttc ttcttctata aaacaatacc
      301 caaagagctc ttcttcttca caattcagat ttcaatttct caaaatctta aaaactttct
      361 ctcaattctc tctaccgtga tcaaggtaaa tttctgtgtt ccttattctc tcaaaatctt
      421 cgattttgtt ttcgttcgat cccaatttcg tatatgttct ttggtttaga ttctgttaat
      481 cttagatcgt agacgatttt ctgggtttga tcgttagata tcatcttaat tctcgattag
      541 ggtttcatag atatcatccg atttgttcaa ataatttgag ttttgtcgaa taattactct
      601 tcgatttgtg atttctatct agatctggtg ttagtttcta gtttgtgcga tcgaatttgt
      661 cgattaatct gagtttttct gattaacagc accatggcaa aggatgtgga agccgttccc
      721 ggagaaggat ttcagacaag agactatcaa gatccgccac cagctccgtt tattgatgga
      781 gcggagctaa agaagtggtc tttctacaga gcagttatcg cagagttcgt agccactctc
      841 ctcttcttat acatcaccgt tttgacagtc atcggttaca agattcagtc cgatactgat
      901 gccggtggcg tagattgcgg cggagttgga atcctcggta tcgcttgggc ctttggtggt
      961 atgatcttca tcctcgtcta ctgcaccgcc ggtatctctg gtggtcacat taacccagcg
     1021 gtgacatttg ggctattctt ggcacgtaaa gtgtcgttac ctagggccct attgtacata
     1081 atcgctcagt gtttgggtgc gatttgtgga gttggttttg tcaaagcctt ccaaagctct
     1141 tactacaccc gttacggagg tggagccaac tctctagccg atggctacag cacagggacc
     1201 ggtctagccg cagagatcat tggtactttc gttcttgtct acaccgtatt ctctgccact
     1261 gaccccaaac gtagtgccag agactcccac gttccggtgt tggcgccact tccaatcgga
     1321 tttgccgtgt tcatggtaca tttggctacc attcccatta ccggaaccgg aattaacccg
     1381 gcaaggagtt tcggagctgc cgtaatctac aacaagagca agccatggga tgaccactgg
     1441 atattttggg ttggaccatt cattggagct gcgatagctg cattctacca ccaattcgtt
     1501 ctaagagctt caggttctaa gtctcttgga tcattcagaa gtgctgccaa cgtcggatcc
     1561 atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag
     1621 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
     1681 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
     1741 ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac
     1801 cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc
     1861 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac
     1921 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta
     1981 atgcagaata agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc
     2041 gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct
     2101 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc
     2161 aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa
     2221 cgcgccgagg gccgccactc caccggcggc atggaggagc tgtacaagta aagcggccgc
     2281 ccggctgcag atcgttcaaa catttggcaa taaagtttct taagattgaa tcctgttgcc
     2341 ggtcttgcga tgattatcat ataatttctg ttgaattacg ttaagcatgt aataattaac
     2401 atgtaatgca tgacgttatt tatgagatgg gtttttatga ttagagtccc gcaattatac
     2461 atttaatacg cgatagaaaa caaaatatag cgcgcaaact aggataaatt atcgcgcgcg
     2521 gtgtcatcta tgttactaga tcggggcgcg ccctcgagga agacgtaatg atagaaaagt
     2581 tggctgcggc cgcgaaactc tatcattgat agagtgaact ctatcattga tagagtcaac
     2641 tctatcattg atagagtcaa ctctatcatt gatagagtgt actctatcat tgatagagta
     2701 gactctatca ttgatagagt cgggtctaag ctagctaggc cacccacaat cccactatcc
     2761 tttcgcaaga cccttcctct atataaggaa gttcatttca tttggagaga acacggggga
     2821 ctctagacac catggtgagt aaaggagaag aacttttcac tggagttgtc ccaattcttg
     2881 ttgaattaga tggtgatgtt aatgggcaca aattttctgt cagtggagag ggtgaaggtg
     2941 atgcaacata cggaaaactt acccttaaat ttatttgcac tactggaaaa ctacctgttc
     3001 catggccaac cctggtcacc accctgacct acgtaagttt ctgcttctac ctttgatata
     3061 tatataataa ttatcattaa ttagtagtaa tataatattt caaatatttt tttcaaaata
     3121 aaagaatgta gtatatagca attgcttttc tgtagtttat aagtgtgtat attttaattt
     3181 ataacttttc taatatatga ccaaaatttg ttgatgtgca gggcgtgcag tgcttctccc
     3241 gttaccctga tcatatgaag cggcacgact tcttcaagag cgccatgcct gagggatacg
     3301 tgcaggagag gaccatcttc ttcaaggacg acgggaacta caagacacgt gctgaagtca
     3361 agtttgaggg tgacaccctc gtcaacagga tcgagcttaa gggaatcgat ttcaaggagg
     3421 acggaaacat cctcggccac aagttggaat acaactacaa ctcccacaac gtatacatca
     3481 tggccgacaa gcaaaagaac ggcatcaaag ccaacttcaa gacccgccac aacatcgagg
     3541 acggcggcgt gcaactcgct gatcattatc aacaaaatac tccaattggc gatggccctg
     3601 tccttttacc agacaaccat tacctgtcca cacaatctgc cctttcgaaa gatcccaacg
     3661 aaaagagaga tcacatggtc cttcttgagt ttgtaacagc tgctgggatt acacatggca
     3721 tggatgaact atacaaataa tgtcaagcca ttctctcgca gatgatgttc actttgtgtt
     3781 ttacttcctt tatgcattca cagcaataaa agaaagaaat ctccatcgct tttggttttc
     3841 ttctctgtct taagttagtc gttttcgtgt ctaatctatt acttatcatt gtaatagact
     3901 cttcttctat tgagatttga atataaacta aaacacattc cattttactg tgttctcaac
     3961 attcagaatg caaacggact aaccgtagtg actcggcttg agtcttcggg ctagagcgga
     4021 ctagttctag agcggccgcc accgcggtgg agctccagct tttgttccct ttagtgaggg
     4081 ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     4141 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4201 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4261 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4321 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4381 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4441 ggaaagaaca tgaaggcctt gacaggatat attggcgggt aaactaagtc gctgtatgtg
     4501 tttgtttgag atctcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4561 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4621 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4681 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4741 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4801 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4861 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4921 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4981 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgtgctctg
     5041 ctgaagccag ttaccttcgg aagaagagtt ggtagctctt gatccggcaa acaaaccacc
     5101 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5161 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5221 taagggattt tggtcatgaa ttcttgttag acattatttg ccgactacct tggtgatctc
     5281 gcctttcacg tagtgaacaa attcttccaa ctgatctgcg cgcgaggcca agcgatcttc
     5341 ttgtccaaga taagcctgcc tagcttcaag tatgacgggc tgatactggg ccggcaggcg
     5401 ctccattgcc cagtcggcag cgacatcctt cggcgcgatt ttgccggtta ctgcgctgta
     5461 ccaaatgcgg gacaacgtaa gcactacatt tcgctcatcg ccagcccagt cgggcggcga
     5521 gttccatagc gttaaggttt catttagcgc ctcaaataga tcctgttcag gaaccggatc
     5581 aaagagttcc tccgccgctg gacctaccaa ggcaacgcta tgttctcttg cttttgtcag
     5641 caagatagcc agatcaatgt cgatcgtggc tggctcgaag atacctgcaa gaatgtcatt
     5701 gcgctgccat tctccaaatt gcagttcgcg cttagctgga taacgccacg gaatgatgtc
     5761 gtcgtgcaca acaatggtga cttctacagc gcggagaatc tcgctctctc caggggaagc
     5821 cgaagtttcc aaaaggtcgt tgatcaaagc tcgccgcgtt gtttcatcaa gccttacggt
     5881 caccgtaacc agcaaatcaa tatcactgtg tggcttcagg ccgccatcca ctgcggagcc
     5941 gtacaaatgt acggccagca acgtcggttc gagatggcgc tcgatgacgc caactacctc
     6001 tgatagttga gtcgatactt cggcgatcac cgcttccctc atgatgttta actcctgaat
     6061 taagccgcgc cgcgaagcgg tgtcggcttg aatgaattgt taggcgtcat cctgtgctcc
     6121 cgagaaccag taccagtaca tcgctgtttc gttcgagact tgaggtctag ttttatacgt
     6181 gaacaggtca atgccgccga gagtaaagcc acattttgcg tacaaattgc aggcaggtac
     6241 attgttcgtt tgtgccatga tgatatattt ttatcttgtg caatgtaaca tcagagattt
     6301 tgagacacaa cgtggctttg ttgaataaat cgaacttttg ctgagttgaa ggatcagatc
     6361 acgcatcttc ccgacaacgc agaccgttcc gtggcaaagc aaaagttcaa aatcaccaac
     6421 tggtccacct acaacaaagc tctcatcaac cgtggctccc tcactttctg gctggatgat
     6481 ggggcgattc aggagatccc catccaacag cccgccgtcg agcgggcttt tttatccccg
     6541 gaagcctgtg gatagagggt agttatccac gtgaaaccgc taatgccccg caaagccttg
     6601 attcacgggg ctttccggcc cgctccaaaa actatccacg tgaaatcgct aatcagggta
     6661 cgtgaaatcg ctaatcggag tacgtgaaat cgctaataag gtcacgtgaa atcgctaatc
     6721 aaaaaggcac gtgagaacgc taatagccct ttcagatcaa cagcttgcaa acacccctcg
     6781 ctccggcaag tagttacagc aagtagtatg ttcaattagc ttttcaatta tgaatatata
     6841 tatcaattat tggtcgccct tggcttgtgg acaatgcgct acgcgcaccg gctccgcccg
     6901 tggacaaccg caagcggttg cccaccgtcg agcgcctttg cccacaaccc ggcggccggc
     6961 cgcaacagat cgttttataa attttttttt ttgaaaaaga aaaagcccga aaggcggcaa
     7021 cctctcgggc ttctggattt ccgatcc
//
