LOCUS       sequence-387069-        7107 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants 
            (proUBQ10::PIP2A-mCherry-NOSt_6XIcaRO-min35S::GFPint-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7107)
  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 7107)
  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: JAB2178
FEATURES             Location/Qualifiers
     source          1..7107
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1540..2250
                     /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(1685..1703)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      2269..2521
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        2804..2850
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     primer_bind     complement(2904..2932)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          3072..3260
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     3716..3736
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4107..4127)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4107..4125)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4146..4162)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4146..4162)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4159..4181)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4170..4186
                     /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(4194..4200,4201..4218,4219..4224))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4239..4260
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4377..4394)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4499..4523
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4614..5202)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4694..4713)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5293..6081)
                     /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      6539..6974
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(7102..7107,1..17)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
ORIGIN
        1 gatatattgt ggtgtaacgt taagctttgg ttgtcgacga gtcagtaata aacggcgtca
       61 aagtggttgc agccggcaca cacgagtcgt gtttatcaac tcaaagcaca aatacttttc
      121 ctcaacctaa aaataaggca attagccaaa aacaactttg cgtgtaaaca acgctcaata
      181 cacgtgtcat tttattatta gctattgctt caccgcctta gctttctcgt gacctagtcg
      241 tcctcgtctt ttcttcttct tcttctataa aacaataccc aaagagctct tcttcttcac
      301 aattcagatt tcaatttctc aaaatcttaa aaactttctc tcaattctct ctaccgtgat
      361 caaggtaaat ttctgtgttc cttattctct caaaatcttc gattttgttt tcgttcgatc
      421 ccaatttcgt atatgttctt tggtttagat tctgttaatc ttagatcgta gacgattttc
      481 tgggtttgat cgttagatat catcttaatt ctcgattagg gtttcataga tatcatccga
      541 tttgttcaaa taatttgagt tttgtcgaat aattactctt cgatttgtga tttctatcta
      601 gatctggtgt tagtttctag tttgtgcgat cgaatttgtc gattaatctg agtttttctg
      661 attaacagca ccatggcaaa ggatgtggaa gccgttcccg gagaaggatt tcagacaaga
      721 gactatcaag atccgccacc agctccgttt attgatggag cggagctaaa gaagtggtct
      781 ttctacagag cagttatcgc agagttcgta gccactctcc tcttcttata catcaccgtt
      841 ttgacagtca tcggttacaa gattcagtcc gatactgatg ccggtggcgt agattgcggc
      901 ggagttggaa tcctcggtat cgcttgggcc tttggtggta tgatcttcat cctcgtctac
      961 tgcaccgccg gtatctctgg tggtcacatt aacccagcgg tgacatttgg gctattcttg
     1021 gcacgtaaag tgtcgttacc tagggcccta ttgtacataa tcgctcagtg tttgggtgcg
     1081 atttgtggag ttggttttgt caaagccttc caaagctctt actacacccg ttacggaggt
     1141 ggagccaact ctctagccga tggctacagc acagggaccg gtctagccgc agagatcatt
     1201 ggtactttcg ttcttgtcta caccgtattc tctgccactg accccaaacg tagtgccaga
     1261 gactcccacg ttccggtgtt ggcgccactt ccaatcggat ttgccgtgtt catggtacat
     1321 ttggctacca ttcccattac cggaaccgga attaacccgg caaggagttt cggagctgcc
     1381 gtaatctaca acaagagcaa gccatgggat gaccactgga tattttgggt tggaccattc
     1441 attggagctg cgatagctgc attctaccac caattcgttc taagagcttc aggttctaag
     1501 tctcttggat cattcagaag tgctgccaac gtcggatcca tggtgagcaa gggcgaggag
     1561 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
     1621 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     1681 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     1741 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
     1801 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     1861 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     1921 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaataa gaccatgggc
     1981 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
     2041 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
     2101 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc
     2161 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     2221 accggcggca tggaggagct gtacaagtaa agcggccgcc cggctgcaga tcgttcaaac
     2281 atttggcaat aaagtttctt aagattgaat cctgttgccg gtcttgcgat gattatcata
     2341 taatttctgt tgaattacgt taagcatgta ataattaaca tgtaatgcat gacgttattt
     2401 atgagatggg tttttatgat tagagtcccg caattataca tttaatacgc gatagaaaac
     2461 aaaatatagc gcgcaaacta ggataaatta tcgcgcgcgg tgtcatctat gttactagat
     2521 cggggcgcgc cctcgaggaa gacgtaatga tagaaaagtt ggctgcggcc gcgaattcac
     2581 ctacctttcg ttaggttagg ttgtgattca cctacctttc gttaggttag gttgtcattc
     2641 acctaccttt cgttaggtta ggttgtcatt cacctacctt tcgttaggtt aggttgtgtt
     2701 tcacctacct ttcgttaggt taggttgtag ttcacctacc tttcgttagg ttaggttgtc
     2761 gggtctaagc tagctaggcc acccacaatc ccactatcct ttcgcaagac ccttcctcta
     2821 tataaggaag ttcatttcat ttggagagaa cacgggggac tctagacacc atggtgagta
     2881 aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat ggtgatgtta
     2941 atgggcacaa attttctgtc agtggagagg gtgaaggtga tgcaacatac ggaaaactta
     3001 cccttaaatt tatttgcact actggaaaac tacctgttcc atggccaacc ctggtcacca
     3061 ccctgaccta cgtaagtttc tgcttctacc tttgatatat atataataat tatcattaat
     3121 tagtagtaat ataatatttc aaatattttt ttcaaaataa aagaatgtag tatatagcaa
     3181 ttgcttttct gtagtttata agtgtgtata ttttaattta taacttttct aatatatgac
     3241 caaaatttgt tgatgtgcag ggcgtgcagt gcttctcccg ttaccctgat catatgaagc
     3301 ggcacgactt cttcaagagc gccatgcctg agggatacgt gcaggagagg accatcttct
     3361 tcaaggacga cgggaactac aagacacgtg ctgaagtcaa gtttgagggt gacaccctcg
     3421 tcaacaggat cgagcttaag ggaatcgatt tcaaggagga cggaaacatc ctcggccaca
     3481 agttggaata caactacaac tcccacaacg tatacatcat ggccgacaag caaaagaacg
     3541 gcatcaaagc caacttcaag acccgccaca acatcgagga cggcggcgtg caactcgctg
     3601 atcattatca acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt
     3661 acctgtccac acaatctgcc ctttcgaaag atcccaacga aaagagagat cacatggtcc
     3721 ttcttgagtt tgtaacagct gctgggatta cacatggcat ggatgaacta tacaaataat
     3781 gtcaagccat tctctcgcag atgatgttca ctttgtgttt tacttccttt atgcattcac
     3841 agcaataaaa gaaagaaatc tccatcgctt ttggttttct tctctgtctt aagttagtcg
     3901 ttttcgtgtc taatctatta cttatcattg taatagactc ttcttctatt gagatttgaa
     3961 tataaactaa aacacattcc attttactgt gttctcaaca ttcagaatgc aaacggacta
     4021 accgtagtga ctcggcttga gtcttcgggc tagagcggac tagttctaga gcggccgcca
     4081 ccgcggtgga gctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
     4141 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4201 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4261 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4321 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4381 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4441 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gaaggccttg
     4501 acaggatata ttggcgggta aactaagtcg ctgtatgtgt ttgtttgaga tctcatgtga
     4561 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4621 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4681 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4741 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4801 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4861 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4921 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4981 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5041 ggctacacta gaagaacagt atttggtatc tgtgctctgc tgaagccagt taccttcgga
     5101 agaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5161 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5221 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaat
     5281 tcttgttaga cattatttgc cgactacctt ggtgatctcg cctttcacgt agtgaacaaa
     5341 ttcttccaac tgatctgcgc gcgaggccaa gcgatcttct tgtccaagat aagcctgcct
     5401 agcttcaagt atgacgggct gatactgggc cggcaggcgc tccattgccc agtcggcagc
     5461 gacatccttc ggcgcgattt tgccggttac tgcgctgtac caaatgcggg acaacgtaag
     5521 cactacattt cgctcatcgc cagcccagtc gggcggcgag ttccatagcg ttaaggtttc
     5581 atttagcgcc tcaaatagat cctgttcagg aaccggatca aagagttcct ccgccgctgg
     5641 acctaccaag gcaacgctat gttctcttgc ttttgtcagc aagatagcca gatcaatgtc
     5701 gatcgtggct ggctcgaaga tacctgcaag aatgtcattg cgctgccatt ctccaaattg
     5761 cagttcgcgc ttagctggat aacgccacgg aatgatgtcg tcgtgcacaa caatggtgac
     5821 ttctacagcg cggagaatct cgctctctcc aggggaagcc gaagtttcca aaaggtcgtt
     5881 gatcaaagct cgccgcgttg tttcatcaag ccttacggtc accgtaacca gcaaatcaat
     5941 atcactgtgt ggcttcaggc cgccatccac tgcggagccg tacaaatgta cggccagcaa
     6001 cgtcggttcg agatggcgct cgatgacgcc aactacctct gatagttgag tcgatacttc
     6061 ggcgatcacc gcttccctca tgatgtttaa ctcctgaatt aagccgcgcc gcgaagcggt
     6121 gtcggcttga atgaattgtt aggcgtcatc ctgtgctccc gagaaccagt accagtacat
     6181 cgctgtttcg ttcgagactt gaggtctagt tttatacgtg aacaggtcaa tgccgccgag
     6241 agtaaagcca cattttgcgt acaaattgca ggcaggtaca ttgttcgttt gtgccatgat
     6301 gatatatttt tatcttgtgc aatgtaacat cagagatttt gagacacaac gtggctttgt
     6361 tgaataaatc gaacttttgc tgagttgaag gatcagatca cgcatcttcc cgacaacgca
     6421 gaccgttccg tggcaaagca aaagttcaaa atcaccaact ggtccaccta caacaaagct
     6481 ctcatcaacc gtggctccct cactttctgg ctggatgatg gggcgattca ggagatcccc
     6541 atccaacagc ccgccgtcga gcgggctttt ttatccccgg aagcctgtgg atagagggta
     6601 gttatccacg tgaaaccgct aatgccccgc aaagccttga ttcacggggc tttccggccc
     6661 gctccaaaaa ctatccacgt gaaatcgcta atcagggtac gtgaaatcgc taatcggagt
     6721 acgtgaaatc gctaataagg tcacgtgaaa tcgctaatca aaaaggcacg tgagaacgct
     6781 aatagccctt tcagatcaac agcttgcaaa cacccctcgc tccggcaagt agttacagca
     6841 agtagtatgt tcaattagct tttcaattat gaatatatat atcaattatt ggtcgccctt
     6901 ggcttgtgga caatgcgcta cgcgcaccgg ctccgcccgt ggacaaccgc aagcggttgc
     6961 ccaccgtcga gcgcctttgc ccacaacccg gcggccggcc gcaacagatc gttttataaa
     7021 tttttttttt tgaaaaagaa aaagcccgaa aggcggcaac ctctcgggct tctggatttc
     7081 cgatccccgg aattagactc ttggcag
//
