LOCUS       sequence-389461-        7059 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants 
            (proUBQ10::PIP2A-mCherry-NOSt_6XLacIO-min35S::GFPint-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7059)
  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 7059)
  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: JAB2493
FEATURES             Location/Qualifiers
     source          1..7059
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    35..57
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     CDS             1580..2290
                     /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(1725..1743)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      2309..2561
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    2618..2634
                     /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)."
     protein_bind    2641..2657
                     /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)."
     protein_bind    2660..2683
                     /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)."
     protein_bind    2687..2703
                     /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)."
     protein_bind    2710..2726
                     /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)."
     protein_bind    2730..2752
                     /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        2796..2842
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     primer_bind     complement(2896..2924)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          3064..3252
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     3708..3728
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4099..4119)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4099..4117)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4138..4154)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4138..4154)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4151..4173)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4162..4178
                     /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(4186..4192,4193..4210,4211..4216))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4231..4252
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4369..4386)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4491..4515
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4606..5194)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4686..4705)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5285..6073)
                     /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      6531..6966
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 gcttctggat ttccgatccc cggaattaga ctcttggcag gatatattgt ggtgtaacgt
       61 taagctttgg ttgtcgacga gtcagtaata aacggcgtca aagtggttgc agccggcaca
      121 cacgagtcgt gtttatcaac tcaaagcaca aatacttttc ctcaacctaa aaataaggca
      181 attagccaaa aacaactttg cgtgtaaaca acgctcaata cacgtgtcat tttattatta
      241 gctattgctt caccgcctta gctttctcgt gacctagtcg tcctcgtctt ttcttcttct
      301 tcttctataa aacaataccc aaagagctct tcttcttcac aattcagatt tcaatttctc
      361 aaaatcttaa aaactttctc tcaattctct ctaccgtgat caaggtaaat ttctgtgttc
      421 cttattctct caaaatcttc gattttgttt tcgttcgatc ccaatttcgt atatgttctt
      481 tggtttagat tctgttaatc ttagatcgta gacgattttc tgggtttgat cgttagatat
      541 catcttaatt ctcgattagg gtttcataga tatcatccga tttgttcaaa taatttgagt
      601 tttgtcgaat aattactctt cgatttgtga tttctatcta gatctggtgt tagtttctag
      661 tttgtgcgat cgaatttgtc gattaatctg agtttttctg attaacagca ccatggcaaa
      721 ggatgtggaa gccgttcccg gagaaggatt tcagacaaga gactatcaag atccgccacc
      781 agctccgttt attgatggag cggagctaaa gaagtggtct ttctacagag cagttatcgc
      841 agagttcgta gccactctcc tcttcttata catcaccgtt ttgacagtca tcggttacaa
      901 gattcagtcc gatactgatg ccggtggcgt agattgcggc ggagttggaa tcctcggtat
      961 cgcttgggcc tttggtggta tgatcttcat cctcgtctac tgcaccgccg gtatctctgg
     1021 tggtcacatt aacccagcgg tgacatttgg gctattcttg gcacgtaaag tgtcgttacc
     1081 tagggcccta ttgtacataa tcgctcagtg tttgggtgcg atttgtggag ttggttttgt
     1141 caaagccttc caaagctctt actacacccg ttacggaggt ggagccaact ctctagccga
     1201 tggctacagc acagggaccg gtctagccgc agagatcatt ggtactttcg ttcttgtcta
     1261 caccgtattc tctgccactg accccaaacg tagtgccaga gactcccacg ttccggtgtt
     1321 ggcgccactt ccaatcggat ttgccgtgtt catggtacat ttggctacca ttcccattac
     1381 cggaaccgga attaacccgg caaggagttt cggagctgcc gtaatctaca acaagagcaa
     1441 gccatgggat gaccactgga tattttgggt tggaccattc attggagctg cgatagctgc
     1501 attctaccac caattcgttc taagagcttc aggttctaag tctcttggat cattcagaag
     1561 tgctgccaac gtcggatcca tggtgagcaa gggcgaggag gataacatgg ccatcatcaa
     1621 ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat
     1681 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
     1741 caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc
     1801 caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga
     1861 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca
     1921 ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt
     1981 cccctccgac ggccccgtaa tgcagaataa gaccatgggc tgggaggcct cctccgagcg
     2041 gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga
     2101 cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct
     2161 gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac
     2221 catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca tggaggagct
     2281 gtacaagtaa agcggccgcc cggctgcaga tcgttcaaac atttggcaat aaagtttctt
     2341 aagattgaat cctgttgccg gtcttgcgat gattatcata taatttctgt tgaattacgt
     2401 taagcatgta ataattaaca tgtaatgcat gacgttattt atgagatggg tttttatgat
     2461 tagagtcccg caattataca tttaatacgc gatagaaaac aaaatatagc gcgcaaacta
     2521 ggataaatta tcgcgcgcgg tgtcatctat gttactagat cggggcgcgc cctcgaggaa
     2581 gacgtaatga tagaaaagtt ggctgcggcc gcgaaaattg tgagcggata acaattgaaa
     2641 ttgtgagcgg ataacaattg gaattgtgag cggataacaa ttcaaattgt gagcggataa
     2701 caattgtaat tgtgagcgga taacaattag aattgtgagc ggataacaat tcgggtctaa
     2761 gctagctagg ccacccacaa tcccactatc ctttcgcaag acccttcctc tatataagga
     2821 agttcatttc atttggagag aacacggggg actctagaca ccatggtgag taaaggagaa
     2881 gaacttttca ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac
     2941 aaattttctg tcagtggaga gggtgaaggt gatgcaacat acggaaaact tacccttaaa
     3001 tttatttgca ctactggaaa actacctgtt ccatggccaa ccctggtcac caccctgacc
     3061 tacgtaagtt tctgcttcta cctttgatat atatataata attatcatta attagtagta
     3121 atataatatt tcaaatattt ttttcaaaat aaaagaatgt agtatatagc aattgctttt
     3181 ctgtagttta taagtgtgta tattttaatt tataactttt ctaatatatg accaaaattt
     3241 gttgatgtgc agggcgtgca gtgcttctcc cgttaccctg atcatatgaa gcggcacgac
     3301 ttcttcaaga gcgccatgcc tgagggatac gtgcaggaga ggaccatctt cttcaaggac
     3361 gacgggaact acaagacacg tgctgaagtc aagtttgagg gtgacaccct cgtcaacagg
     3421 atcgagctta agggaatcga tttcaaggag gacggaaaca tcctcggcca caagttggaa
     3481 tacaactaca actcccacaa cgtatacatc atggccgaca agcaaaagaa cggcatcaaa
     3541 gccaacttca agacccgcca caacatcgag gacggcggcg tgcaactcgc tgatcattat
     3601 caacaaaata ctccaattgg cgatggccct gtccttttac cagacaacca ttacctgtcc
     3661 acacaatctg ccctttcgaa agatcccaac gaaaagagag atcacatggt ccttcttgag
     3721 tttgtaacag ctgctgggat tacacatggc atggatgaac tatacaaata atgtcaagcc
     3781 attctctcgc agatgatgtt cactttgtgt tttacttcct ttatgcattc acagcaataa
     3841 aagaaagaaa tctccatcgc ttttggtttt cttctctgtc ttaagttagt cgttttcgtg
     3901 tctaatctat tacttatcat tgtaatagac tcttcttcta ttgagatttg aatataaact
     3961 aaaacacatt ccattttact gtgttctcaa cattcagaat gcaaacggac taaccgtagt
     4021 gactcggctt gagtcttcgg gctagagcgg actagttcta gagcggccgc caccgcggtg
     4081 gagctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc
     4141 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4201 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4261 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4321 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4381 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4441 acggttatcc acagaatcag gggataacgc aggaaagaac atgaaggcct tgacaggata
     4501 tattggcggg taaactaagt cgctgtatgt gtttgtttga gatctcatgt gagcaaaagg
     4561 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4621 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4681 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4741 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4801 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4861 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4921 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4981 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5041 tagaagaaca gtatttggta tctgtgctct gctgaagcca gttaccttcg gaagaagagt
     5101 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5161 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5221 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga attcttgtta
     5281 gacattattt gccgactacc ttggtgatct cgcctttcac gtagtgaaca aattcttcca
     5341 actgatctgc gcgcgaggcc aagcgatctt cttgtccaag ataagcctgc ctagcttcaa
     5401 gtatgacggg ctgatactgg gccggcaggc gctccattgc ccagtcggca gcgacatcct
     5461 tcggcgcgat tttgccggtt actgcgctgt accaaatgcg ggacaacgta agcactacat
     5521 ttcgctcatc gccagcccag tcgggcggcg agttccatag cgttaaggtt tcatttagcg
     5581 cctcaaatag atcctgttca ggaaccggat caaagagttc ctccgccgct ggacctacca
     5641 aggcaacgct atgttctctt gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg
     5701 ctggctcgaa gatacctgca agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc
     5761 gcttagctgg ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg acttctacag
     5821 cgcggagaat ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag
     5881 ctcgccgcgt tgtttcatca agccttacgg tcaccgtaac cagcaaatca atatcactgt
     5941 gtggcttcag gccgccatcc actgcggagc cgtacaaatg tacggccagc aacgtcggtt
     6001 cgagatggcg ctcgatgacg ccaactacct ctgatagttg agtcgatact tcggcgatca
     6061 ccgcttccct catgatgttt aactcctgaa ttaagccgcg ccgcgaagcg gtgtcggctt
     6121 gaatgaattg ttaggcgtca tcctgtgctc ccgagaacca gtaccagtac atcgctgttt
     6181 cgttcgagac ttgaggtcta gttttatacg tgaacaggtc aatgccgccg agagtaaagc
     6241 cacattttgc gtacaaattg caggcaggta cattgttcgt ttgtgccatg atgatatatt
     6301 tttatcttgt gcaatgtaac atcagagatt ttgagacaca acgtggcttt gttgaataaa
     6361 tcgaactttt gctgagttga aggatcagat cacgcatctt cccgacaacg cagaccgttc
     6421 cgtggcaaag caaaagttca aaatcaccaa ctggtccacc tacaacaaag ctctcatcaa
     6481 ccgtggctcc ctcactttct ggctggatga tggggcgatt caggagatcc ccatccaaca
     6541 gcccgccgtc gagcgggctt ttttatcccc ggaagcctgt ggatagaggg tagttatcca
     6601 cgtgaaaccg ctaatgcccc gcaaagcctt gattcacggg gctttccggc ccgctccaaa
     6661 aactatccac gtgaaatcgc taatcagggt acgtgaaatc gctaatcgga gtacgtgaaa
     6721 tcgctaataa ggtcacgtga aatcgctaat caaaaaggca cgtgagaacg ctaatagccc
     6781 tttcagatca acagcttgca aacacccctc gctccggcaa gtagttacag caagtagtat
     6841 gttcaattag cttttcaatt atgaatatat atatcaatta ttggtcgccc ttggcttgtg
     6901 gacaatgcgc tacgcgcacc ggctccgccc gtggacaacc gcaagcggtt gcccaccgtc
     6961 gagcgccttt gcccacaacc cggcggccgg ccgcaacaga tcgttttata aatttttttt
     7021 tttgaaaaag aaaaagcccg aaaggcggca acctctcgg
//
