LOCUS       sequence-387067-        7065 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants 
            (proUBQ10::PIP2A-mCherry-NOSt_6XAmtRO-min35S::GFPint-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7065)
  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 7065)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: JAB2177
FEATURES             Location/Qualifiers
     source          1..7065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    10..32
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     CDS             1555..2265
                     /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(1700..1718)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      2284..2536
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        2777..2823
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     primer_bind     complement(2877..2905)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          3045..3233
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     3689..3709
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4080..4100)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4080..4098)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4119..4135)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4119..4135)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4132..4154)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4143..4159
                     /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(4167..4173,4174..4191,4192..4197))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4212..4233
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4350..4367)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4472..4496
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4587..5175)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4667..4686)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5266..6054)
                     /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      6512..6947
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 ttagactctt ggcaggatat attgtggtgt aacgttaagc tttggttgtc gacgagtcag
       61 taataaacgg cgtcaaagtg gttgcagccg gcacacacga gtcgtgttta tcaactcaaa
      121 gcacaaatac ttttcctcaa cctaaaaata aggcaattag ccaaaaacaa ctttgcgtgt
      181 aaacaacgct caatacacgt gtcattttat tattagctat tgcttcaccg ccttagcttt
      241 ctcgtgacct agtcgtcctc gtcttttctt cttcttcttc tataaaacaa tacccaaaga
      301 gctcttcttc ttcacaattc agatttcaat ttctcaaaat cttaaaaact ttctctcaat
      361 tctctctacc gtgatcaagg taaatttctg tgttccttat tctctcaaaa tcttcgattt
      421 tgttttcgtt cgatcccaat ttcgtatatg ttctttggtt tagattctgt taatcttaga
      481 tcgtagacga ttttctgggt ttgatcgtta gatatcatct taattctcga ttagggtttc
      541 atagatatca tccgatttgt tcaaataatt tgagttttgt cgaataatta ctcttcgatt
      601 tgtgatttct atctagatct ggtgttagtt tctagtttgt gcgatcgaat ttgtcgatta
      661 atctgagttt ttctgattaa cagcaccatg gcaaaggatg tggaagccgt tcccggagaa
      721 ggatttcaga caagagacta tcaagatccg ccaccagctc cgtttattga tggagcggag
      781 ctaaagaagt ggtctttcta cagagcagtt atcgcagagt tcgtagccac tctcctcttc
      841 ttatacatca ccgttttgac agtcatcggt tacaagattc agtccgatac tgatgccggt
      901 ggcgtagatt gcggcggagt tggaatcctc ggtatcgctt gggcctttgg tggtatgatc
      961 ttcatcctcg tctactgcac cgccggtatc tctggtggtc acattaaccc agcggtgaca
     1021 tttgggctat tcttggcacg taaagtgtcg ttacctaggg ccctattgta cataatcgct
     1081 cagtgtttgg gtgcgatttg tggagttggt tttgtcaaag ccttccaaag ctcttactac
     1141 acccgttacg gaggtggagc caactctcta gccgatggct acagcacagg gaccggtcta
     1201 gccgcagaga tcattggtac tttcgttctt gtctacaccg tattctctgc cactgacccc
     1261 aaacgtagtg ccagagactc ccacgttccg gtgttggcgc cacttccaat cggatttgcc
     1321 gtgttcatgg tacatttggc taccattccc attaccggaa ccggaattaa cccggcaagg
     1381 agtttcggag ctgccgtaat ctacaacaag agcaagccat gggatgacca ctggatattt
     1441 tgggttggac cattcattgg agctgcgata gctgcattct accaccaatt cgttctaaga
     1501 gcttcaggtt ctaagtctct tggatcattc agaagtgctg ccaacgtcgg atccatggtg
     1561 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
     1621 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     1681 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
     1741 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
     1801 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     1861 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     1921 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     1981 aataagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
     2041 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
     2101 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
     2161 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
     2221 gagggccgcc actccaccgg cggcatggag gagctgtaca agtaaagcgg ccgcccggct
     2281 gcagatcgtt caaacatttg gcaataaagt ttcttaagat tgaatcctgt tgccggtctt
     2341 gcgatgatta tcatataatt tctgttgaat tacgttaagc atgtaataat taacatgtaa
     2401 tgcatgacgt tatttatgag atgggttttt atgattagag tcccgcaatt atacatttaa
     2461 tacgcgatag aaaacaaaat atagcgcgca aactaggata aattatcgcg cgcggtgtca
     2521 tctatgttac tagatcgggg cgcgccctcg aggaagacgt aatgatagaa aagttggctg
     2581 cggccgcgaa tttctatcga tctatagata atgatttcta tcgatctata gataatcatt
     2641 tctatcgatc tatagataat catttctatc gatctataga taatgttttc tatcgatcta
     2701 tagataatag tttctatcga tctatagata atcgggtcta agctagctag gccacccaca
     2761 atcccactat cctttcgcaa gacccttcct ctatataagg aagttcattt catttggaga
     2821 gaacacgggg gactctagac accatggtga gtaaaggaga agaacttttc actggagttg
     2881 tcccaattct tgttgaatta gatggtgatg ttaatgggca caaattttct gtcagtggag
     2941 agggtgaagg tgatgcaaca tacggaaaac ttacccttaa atttatttgc actactggaa
     3001 aactacctgt tccatggcca accctggtca ccaccctgac ctacgtaagt ttctgcttct
     3061 acctttgata tatatataat aattatcatt aattagtagt aatataatat ttcaaatatt
     3121 tttttcaaaa taaaagaatg tagtatatag caattgcttt tctgtagttt ataagtgtgt
     3181 atattttaat ttataacttt tctaatatat gaccaaaatt tgttgatgtg cagggcgtgc
     3241 agtgcttctc ccgttaccct gatcatatga agcggcacga cttcttcaag agcgccatgc
     3301 ctgagggata cgtgcaggag aggaccatct tcttcaagga cgacgggaac tacaagacac
     3361 gtgctgaagt caagtttgag ggtgacaccc tcgtcaacag gatcgagctt aagggaatcg
     3421 atttcaagga ggacggaaac atcctcggcc acaagttgga atacaactac aactcccaca
     3481 acgtatacat catggccgac aagcaaaaga acggcatcaa agccaacttc aagacccgcc
     3541 acaacatcga ggacggcggc gtgcaactcg ctgatcatta tcaacaaaat actccaattg
     3601 gcgatggccc tgtcctttta ccagacaacc attacctgtc cacacaatct gccctttcga
     3661 aagatcccaa cgaaaagaga gatcacatgg tccttcttga gtttgtaaca gctgctggga
     3721 ttacacatgg catggatgaa ctatacaaat aatgtcaagc cattctctcg cagatgatgt
     3781 tcactttgtg ttttacttcc tttatgcatt cacagcaata aaagaaagaa atctccatcg
     3841 cttttggttt tcttctctgt cttaagttag tcgttttcgt gtctaatcta ttacttatca
     3901 ttgtaataga ctcttcttct attgagattt gaatataaac taaaacacat tccattttac
     3961 tgtgttctca acattcagaa tgcaaacgga ctaaccgtag tgactcggct tgagtcttcg
     4021 ggctagagcg gactagttct agagcggccg ccaccgcggt ggagctccag cttttgttcc
     4081 ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
     4141 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4201 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4261 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4321 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4381 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4441 ggggataacg caggaaagaa catgaaggcc ttgacaggat atattggcgg gtaaactaag
     4501 tcgctgtatg tgtttgtttg agatctcatg tgagcaaaag gccagcaaaa ggccaggaac
     4561 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4621 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4681 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4741 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4801 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4861 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4921 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4981 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5041 atctgtgctc tgctgaagcc agttaccttc ggaagaagag ttggtagctc ttgatccggc
     5101 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5161 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5221 gaaaactcac gttaagggat tttggtcatg aattcttgtt agacattatt tgccgactac
     5281 cttggtgatc tcgcctttca cgtagtgaac aaattcttcc aactgatctg cgcgcgaggc
     5341 caagcgatct tcttgtccaa gataagcctg cctagcttca agtatgacgg gctgatactg
     5401 ggccggcagg cgctccattg cccagtcggc agcgacatcc ttcggcgcga ttttgccggt
     5461 tactgcgctg taccaaatgc gggacaacgt aagcactaca tttcgctcat cgccagccca
     5521 gtcgggcggc gagttccata gcgttaaggt ttcatttagc gcctcaaata gatcctgttc
     5581 aggaaccgga tcaaagagtt cctccgccgc tggacctacc aaggcaacgc tatgttctct
     5641 tgcttttgtc agcaagatag ccagatcaat gtcgatcgtg gctggctcga agatacctgc
     5701 aagaatgtca ttgcgctgcc attctccaaa ttgcagttcg cgcttagctg gataacgcca
     5761 cggaatgatg tcgtcgtgca caacaatggt gacttctaca gcgcggagaa tctcgctctc
     5821 tccaggggaa gccgaagttt ccaaaaggtc gttgatcaaa gctcgccgcg ttgtttcatc
     5881 aagccttacg gtcaccgtaa ccagcaaatc aatatcactg tgtggcttca ggccgccatc
     5941 cactgcggag ccgtacaaat gtacggccag caacgtcggt tcgagatggc gctcgatgac
     6001 gccaactacc tctgatagtt gagtcgatac ttcggcgatc accgcttccc tcatgatgtt
     6061 taactcctga attaagccgc gccgcgaagc ggtgtcggct tgaatgaatt gttaggcgtc
     6121 atcctgtgct cccgagaacc agtaccagta catcgctgtt tcgttcgaga cttgaggtct
     6181 agttttatac gtgaacaggt caatgccgcc gagagtaaag ccacattttg cgtacaaatt
     6241 gcaggcaggt acattgttcg tttgtgccat gatgatatat ttttatcttg tgcaatgtaa
     6301 catcagagat tttgagacac aacgtggctt tgttgaataa atcgaacttt tgctgagttg
     6361 aaggatcaga tcacgcatct tcccgacaac gcagaccgtt ccgtggcaaa gcaaaagttc
     6421 aaaatcacca actggtccac ctacaacaaa gctctcatca accgtggctc cctcactttc
     6481 tggctggatg atggggcgat tcaggagatc cccatccaac agcccgccgt cgagcgggct
     6541 tttttatccc cggaagcctg tggatagagg gtagttatcc acgtgaaacc gctaatgccc
     6601 cgcaaagcct tgattcacgg ggctttccgg cccgctccaa aaactatcca cgtgaaatcg
     6661 ctaatcaggg tacgtgaaat cgctaatcgg agtacgtgaa atcgctaata aggtcacgtg
     6721 aaatcgctaa tcaaaaaggc acgtgagaac gctaatagcc ctttcagatc aacagcttgc
     6781 aaacacccct cgctccggca agtagttaca gcaagtagta tgttcaatta gcttttcaat
     6841 tatgaatata tatatcaatt attggtcgcc cttggcttgt ggacaatgcg ctacgcgcac
     6901 cggctccgcc cgtggacaac cgcaagcggt tgcccaccgt cgagcgcctt tgcccacaac
     6961 ccggcggccg gccgcaacag atcgttttat aaattttttt ttttgaaaaa gaaaaagccc
     7021 gaaaggcggc aacctctcgg gcttctggat ttccgatccc cggaa
//
