LOCUS       sequence-391173-        6985 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proUBQ10::PIP2A-mCherry-NOSt_2XPhlFO::GFPint-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6985)
  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 6985)
  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: JAB3605
FEATURES             Location/Qualifiers
     source          1..6985
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    13..35
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     CDS             1558..2268
                     /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(1703..1721)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      2287..2539
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        2700..2746
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     primer_bind     complement(2800..2828)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          2968..3156
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     3612..3632
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4003..4023)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4003..4021)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4042..4058)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4042..4058)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4055..4077)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4066..4082
                     /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(4090..4096,4097..4114,4115..4120))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4135..4156
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4273..4290)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4395..4419
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4510..5098)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4590..4609)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5189..5977)
                     /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      6435..6870
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 gaattagact cttggcagga tatattgtgg tgtaacgtta agctttggtt gtcgacgagt
       61 cagtaataaa cggcgtcaaa gtggttgcag ccggcacaca cgagtcgtgt ttatcaactc
      121 aaagcacaaa tacttttcct caacctaaaa ataaggcaat tagccaaaaa caactttgcg
      181 tgtaaacaac gctcaataca cgtgtcattt tattattagc tattgcttca ccgccttagc
      241 tttctcgtga cctagtcgtc ctcgtctttt cttcttcttc ttctataaaa caatacccaa
      301 agagctcttc ttcttcacaa ttcagatttc aatttctcaa aatcttaaaa actttctctc
      361 aattctctct accgtgatca aggtaaattt ctgtgttcct tattctctca aaatcttcga
      421 ttttgttttc gttcgatccc aatttcgtat atgttctttg gtttagattc tgttaatctt
      481 agatcgtaga cgattttctg ggtttgatcg ttagatatca tcttaattct cgattagggt
      541 ttcatagata tcatccgatt tgttcaaata atttgagttt tgtcgaataa ttactcttcg
      601 atttgtgatt tctatctaga tctggtgtta gtttctagtt tgtgcgatcg aatttgtcga
      661 ttaatctgag tttttctgat taacagcacc atggcaaagg atgtggaagc cgttcccgga
      721 gaaggatttc agacaagaga ctatcaagat ccgccaccag ctccgtttat tgatggagcg
      781 gagctaaaga agtggtcttt ctacagagca gttatcgcag agttcgtagc cactctcctc
      841 ttcttataca tcaccgtttt gacagtcatc ggttacaaga ttcagtccga tactgatgcc
      901 ggtggcgtag attgcggcgg agttggaatc ctcggtatcg cttgggcctt tggtggtatg
      961 atcttcatcc tcgtctactg caccgccggt atctctggtg gtcacattaa cccagcggtg
     1021 acatttgggc tattcttggc acgtaaagtg tcgttaccta gggccctatt gtacataatc
     1081 gctcagtgtt tgggtgcgat ttgtggagtt ggttttgtca aagccttcca aagctcttac
     1141 tacacccgtt acggaggtgg agccaactct ctagccgatg gctacagcac agggaccggt
     1201 ctagccgcag agatcattgg tactttcgtt cttgtctaca ccgtattctc tgccactgac
     1261 cccaaacgta gtgccagaga ctcccacgtt ccggtgttgg cgccacttcc aatcggattt
     1321 gccgtgttca tggtacattt ggctaccatt cccattaccg gaaccggaat taacccggca
     1381 aggagtttcg gagctgccgt aatctacaac aagagcaagc catgggatga ccactggata
     1441 ttttgggttg gaccattcat tggagctgcg atagctgcat tctaccacca attcgttcta
     1501 agagcttcag gttctaagtc tcttggatca ttcagaagtg ctgccaacgt cggatccatg
     1561 gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg
     1621 cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
     1681 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc
     1741 gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc
     1801 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
     1861 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
     1921 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
     1981 cagaataaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc
     2041 ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag
     2101 gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac
     2161 atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc
     2221 gccgagggcc gccactccac cggcggcatg gaggagctgt acaagtaaag cggccgcccg
     2281 gctgcagatc gttcaaacat ttggcaataa agtttcttaa gattgaatcc tgttgccggt
     2341 cttgcgatga ttatcatata atttctgttg aattacgtta agcatgtaat aattaacatg
     2401 taatgcatga cgttatttat gagatgggtt tttatgatta gagtcccgca attatacatt
     2461 taatacgcga tagaaaacaa aatatagcgc gcaaactagg ataaattatc gcgcgcggtg
     2521 tcatctatgt tactagatcg gggcgcgccc tcgaggaaga cgtaatgata gaaaagttgg
     2581 ctgcggccgc gaaatgatac gaaacgtacc gtatcgttaa ggtagatgat acgaaacgta
     2641 ccgtatcgtt aaggtcgggt ctaagctagc taggccaccc acaatcccac tatcctttcg
     2701 caagaccctt cctctatata aggaagttca tttcatttgg agagaacacg ggggactcta
     2761 gacaccatgg tgagtaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa
     2821 ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga aggtgatgca
     2881 acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc tgttccatgg
     2941 ccaaccctgg tcaccaccct gacctacgta agtttctgct tctacctttg atatatatat
     3001 aataattatc attaattagt agtaatataa tatttcaaat atttttttca aaataaaaga
     3061 atgtagtata tagcaattgc ttttctgtag tttataagtg tgtatatttt aatttataac
     3121 ttttctaata tatgaccaaa atttgttgat gtgcagggcg tgcagtgctt ctcccgttac
     3181 cctgatcata tgaagcggca cgacttcttc aagagcgcca tgcctgaggg atacgtgcag
     3241 gagaggacca tcttcttcaa ggacgacggg aactacaaga cacgtgctga agtcaagttt
     3301 gagggtgaca ccctcgtcaa caggatcgag cttaagggaa tcgatttcaa ggaggacgga
     3361 aacatcctcg gccacaagtt ggaatacaac tacaactccc acaacgtata catcatggcc
     3421 gacaagcaaa agaacggcat caaagccaac ttcaagaccc gccacaacat cgaggacggc
     3481 ggcgtgcaac tcgctgatca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt
     3541 ttaccagaca accattacct gtccacacaa tctgcccttt cgaaagatcc caacgaaaag
     3601 agagatcaca tggtccttct tgagtttgta acagctgctg ggattacaca tggcatggat
     3661 gaactataca aataatgtca agccattctc tcgcagatga tgttcacttt gtgttttact
     3721 tcctttatgc attcacagca ataaaagaaa gaaatctcca tcgcttttgg ttttcttctc
     3781 tgtcttaagt tagtcgtttt cgtgtctaat ctattactta tcattgtaat agactcttct
     3841 tctattgaga tttgaatata aactaaaaca cattccattt tactgtgttc tcaacattca
     3901 gaatgcaaac ggactaaccg tagtgactcg gcttgagtct tcgggctaga gcggactagt
     3961 tctagagcgg ccgccaccgc ggtggagctc cagcttttgt tccctttagt gagggttaat
     4021 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4081 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4141 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4201 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4261 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4321 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4381 gaacatgaag gccttgacag gatatattgg cgggtaaact aagtcgctgt atgtgtttgt
     4441 ttgagatctc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4501 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4561 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4621 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4681 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4741 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4801 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4861 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4921 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgtg ctctgctgaa
     4981 gccagttacc ttcggaagaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5041 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5101 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5161 gattttggtc atgaattctt gttagacatt atttgccgac taccttggtg atctcgcctt
     5221 tcacgtagtg aacaaattct tccaactgat ctgcgcgcga ggccaagcga tcttcttgtc
     5281 caagataagc ctgcctagct tcaagtatga cgggctgata ctgggccggc aggcgctcca
     5341 ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc ggttactgcg ctgtaccaaa
     5401 tgcgggacaa cgtaagcact acatttcgct catcgccagc ccagtcgggc ggcgagttcc
     5461 atagcgttaa ggtttcattt agcgcctcaa atagatcctg ttcaggaacc ggatcaaaga
     5521 gttcctccgc cgctggacct accaaggcaa cgctatgttc tcttgctttt gtcagcaaga
     5581 tagccagatc aatgtcgatc gtggctggct cgaagatacc tgcaagaatg tcattgcgct
     5641 gccattctcc aaattgcagt tcgcgcttag ctggataacg ccacggaatg atgtcgtcgt
     5701 gcacaacaat ggtgacttct acagcgcgga gaatctcgct ctctccaggg gaagccgaag
     5761 tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc atcaagcctt acggtcaccg
     5821 taaccagcaa atcaatatca ctgtgtggct tcaggccgcc atccactgcg gagccgtaca
     5881 aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat gacgccaact acctctgata
     5941 gttgagtcga tacttcggcg atcaccgctt ccctcatgat gtttaactcc tgaattaagc
     6001 cgcgccgcga agcggtgtcg gcttgaatga attgttaggc gtcatcctgt gctcccgaga
     6061 accagtacca gtacatcgct gtttcgttcg agacttgagg tctagtttta tacgtgaaca
     6121 ggtcaatgcc gccgagagta aagccacatt ttgcgtacaa attgcaggca ggtacattgt
     6181 tcgtttgtgc catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga
     6241 cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc agatcacgca
     6301 tcttcccgac aacgcagacc gttccgtggc aaagcaaaag ttcaaaatca ccaactggtc
     6361 cacctacaac aaagctctca tcaaccgtgg ctccctcact ttctggctgg atgatggggc
     6421 gattcaggag atccccatcc aacagcccgc cgtcgagcgg gcttttttat ccccggaagc
     6481 ctgtggatag agggtagtta tccacgtgaa accgctaatg ccccgcaaag ccttgattca
     6541 cggggctttc cggcccgctc caaaaactat ccacgtgaaa tcgctaatca gggtacgtga
     6601 aatcgctaat cggagtacgt gaaatcgcta ataaggtcac gtgaaatcgc taatcaaaaa
     6661 ggcacgtgag aacgctaata gccctttcag atcaacagct tgcaaacacc cctcgctccg
     6721 gcaagtagtt acagcaagta gtatgttcaa ttagcttttc aattatgaat atatatatca
     6781 attattggtc gcccttggct tgtggacaat gcgctacgcg caccggctcc gcccgtggac
     6841 aaccgcaagc ggttgcccac cgtcgagcgc ctttgcccac aacccggcgg ccggccgcaa
     6901 cagatcgttt tataaatttt tttttttgaa aaagaaaaag cccgaaaggc ggcaacctct
     6961 cgggcttctg gatttccgat ccccg
//
