LOCUS       sequence-393462-        7644 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proUBQ10::PIP2A-mCherry-NOSt_pro35S::GFPint-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7644)
  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 7644)
  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: JAB3422
FEATURES             Location/Qualifiers
     source          1..7644
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1541..2251
                     /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(1686..1704)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      2270..2522
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        3045..3390
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     3333..3353
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     complement(3442..3470)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          3610..3798
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     4254..4274
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4645..4665)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4645..4663)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4684..4700)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4684..4700)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4697..4719)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4708..4724
                     /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(4732..4738,4739..4756,4757..4762))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4777..4798
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4915..4932)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5037..5061
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5152..5740)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5232..5251)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5831..6619)
                     /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      7077..7512
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(7640..7644,1..18)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
ORIGIN
        1 ggatatattg tggtgtaacg ttaagctttg gttgtcgacg agtcagtaat aaacggcgtc
       61 aaagtggttg cagccggcac acacgagtcg tgtttatcaa ctcaaagcac aaatactttt
      121 cctcaaccta aaaataaggc aattagccaa aaacaacttt gcgtgtaaac aacgctcaat
      181 acacgtgtca ttttattatt agctattgct tcaccgcctt agctttctcg tgacctagtc
      241 gtcctcgtct tttcttcttc ttcttctata aaacaatacc caaagagctc ttcttcttca
      301 caattcagat ttcaatttct caaaatctta aaaactttct ctcaattctc tctaccgtga
      361 tcaaggtaaa tttctgtgtt ccttattctc tcaaaatctt cgattttgtt ttcgttcgat
      421 cccaatttcg tatatgttct ttggtttaga ttctgttaat cttagatcgt agacgatttt
      481 ctgggtttga tcgttagata tcatcttaat tctcgattag ggtttcatag atatcatccg
      541 atttgttcaa ataatttgag ttttgtcgaa taattactct tcgatttgtg atttctatct
      601 agatctggtg ttagtttcta gtttgtgcga tcgaatttgt cgattaatct gagtttttct
      661 gattaacagc accatggcaa aggatgtgga agccgttccc ggagaaggat ttcagacaag
      721 agactatcaa gatccgccac cagctccgtt tattgatgga gcggagctaa agaagtggtc
      781 tttctacaga gcagttatcg cagagttcgt agccactctc ctcttcttat acatcaccgt
      841 tttgacagtc atcggttaca agattcagtc cgatactgat gccggtggcg tagattgcgg
      901 cggagttgga atcctcggta tcgcttgggc ctttggtggt atgatcttca tcctcgtcta
      961 ctgcaccgcc ggtatctctg gtggtcacat taacccagcg gtgacatttg ggctattctt
     1021 ggcacgtaaa gtgtcgttac ctagggccct attgtacata atcgctcagt gtttgggtgc
     1081 gatttgtgga gttggttttg tcaaagcctt ccaaagctct tactacaccc gttacggagg
     1141 tggagccaac tctctagccg atggctacag cacagggacc ggtctagccg cagagatcat
     1201 tggtactttc gttcttgtct acaccgtatt ctctgccact gaccccaaac gtagtgccag
     1261 agactcccac gttccggtgt tggcgccact tccaatcgga tttgccgtgt tcatggtaca
     1321 tttggctacc attcccatta ccggaaccgg aattaacccg gcaaggagtt tcggagctgc
     1381 cgtaatctac aacaagagca agccatggga tgaccactgg atattttggg ttggaccatt
     1441 cattggagct gcgatagctg cattctacca ccaattcgtt ctaagagctt caggttctaa
     1501 gtctcttgga tcattcagaa gtgctgccaa cgtcggatcc atggtgagca agggcgagga
     1561 ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt
     1621 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
     1681 gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc
     1741 ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta
     1801 cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
     1861 cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt
     1921 gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaata agaccatggg
     1981 ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa
     2041 gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa
     2101 ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac
     2161 ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc
     2221 caccggcggc atggaggagc tgtacaagta aagcggccgc ccggctgcag atcgttcaaa
     2281 catttggcaa taaagtttct taagattgaa tcctgttgcc ggtcttgcga tgattatcat
     2341 ataatttctg ttgaattacg ttaagcatgt aataattaac atgtaatgca tgacgttatt
     2401 tatgagatgg gtttttatga ttagagtccc gcaattatac atttaatacg cgatagaaaa
     2461 caaaatatag cgcgcaaact aggataaatt atcgcgcgcg gtgtcatcta tgttactaga
     2521 tcggggcgcg ccctcgagga agacgtaatg ggtccccaga tttgcctttt caatttcaga
     2581 aagaatgcta acccacagat ggttagagag gcttacgcag cagctctcat caagacgatc
     2641 tacccgagca ataatctcca ggaaatcaaa taccttccca agaaggttaa agatgcagtc
     2701 aaaagattca ggactaactg catcaagaac acagagaaag atatatttct caagatcaga
     2761 agtactattc cagtatggac gattcaaggc ttgcttcaca aaccaaggca agtaatagag
     2821 attggagtct ctaaaaaggt agttcccact gaatcaaagg ccatggagtc aaagattcaa
     2881 atagaggacc taacagaact cgccgtaaag actggcgaac agttcataca gagtctctta
     2941 cgactcaatg acaagaagaa aatcttcgtc aacatggtgg agcacgacac acttgtctac
     3001 tccaaaaata tcaaagatac agtctcagaa gaccaaaggg caattgagac ttttcaacaa
     3061 agggtaatat ccggaaacct cctcggattc cattgcccag ctatctgtca ctttattgtg
     3121 aagatagtgg aaaaggaagg tggctcctac aaatgccatc attgcgataa aggaaaggcc
     3181 atcgttgaag atgcctctgc cgacagtggt cccaaagatg gacccccacc cacgaggagc
     3241 atcgtggaaa aagaagacgt tccaaccacg tcttcaaagc aagtggattg atgtgatatc
     3301 tccactgacg taagggatga cgcacaatcc cactatcctt cgcaagaccc ttcctctata
     3361 taaggaagtt catttcattt ggagagaaca cgggggactc tagacaccat ggtgagtaaa
     3421 ggagaagaac ttttcactgg agttgtccca attcttgttg aattagatgg tgatgttaat
     3481 gggcacaaat tttctgtcag tggagagggt gaaggtgatg caacatacgg aaaacttacc
     3541 cttaaattta tttgcactac tggaaaacta cctgttccat ggccaaccct ggtcaccacc
     3601 ctgacctacg taagtttctg cttctacctt tgatatatat ataataatta tcattaatta
     3661 gtagtaatat aatatttcaa atattttttt caaaataaaa gaatgtagta tatagcaatt
     3721 gcttttctgt agtttataag tgtgtatatt ttaatttata acttttctaa tatatgacca
     3781 aaatttgttg atgtgcaggg cgtgcagtgc ttctcccgtt accctgatca tatgaagcgg
     3841 cacgacttct tcaagagcgc catgcctgag ggatacgtgc aggagaggac catcttcttc
     3901 aaggacgacg ggaactacaa gacacgtgct gaagtcaagt ttgagggtga caccctcgtc
     3961 aacaggatcg agcttaaggg aatcgatttc aaggaggacg gaaacatcct cggccacaag
     4021 ttggaataca actacaactc ccacaacgta tacatcatgg ccgacaagca aaagaacggc
     4081 atcaaagcca acttcaagac ccgccacaac atcgaggacg gcggcgtgca actcgctgat
     4141 cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac
     4201 ctgtccacac aatctgccct ttcgaaagat cccaacgaaa agagagatca catggtcctt
     4261 cttgagtttg taacagctgc tgggattaca catggcatgg atgaactata caaataatgt
     4321 caagccattc tctcgcagat gatgttcact ttgtgtttta cttcctttat gcattcacag
     4381 caataaaaga aagaaatctc catcgctttt ggttttcttc tctgtcttaa gttagtcgtt
     4441 ttcgtgtcta atctattact tatcattgta atagactctt cttctattga gatttgaata
     4501 taaactaaaa cacattccat tttactgtgt tctcaacatt cagaatgcaa acggactaac
     4561 cgtagtgact cggcttgagt cttcgggcta gagcggacta gttctagagc ggccgccacc
     4621 gcggtggagc tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
     4681 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4741 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4801 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4861 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4921 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4981 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatga aggccttgac
     5041 aggatatatt ggcgggtaaa ctaagtcgct gtatgtgttt gtttgagatc tcatgtgagc
     5101 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5161 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5221 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5281 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5341 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5401 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5461 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5521 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5581 ctacactaga agaacagtat ttggtatctg tgctctgctg aagccagtta ccttcggaag
     5641 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5701 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5761 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgaattc
     5821 ttgttagaca ttatttgccg actaccttgg tgatctcgcc tttcacgtag tgaacaaatt
     5881 cttccaactg atctgcgcgc gaggccaagc gatcttcttg tccaagataa gcctgcctag
     5941 cttcaagtat gacgggctga tactgggccg gcaggcgctc cattgcccag tcggcagcga
     6001 catccttcgg cgcgattttg ccggttactg cgctgtacca aatgcgggac aacgtaagca
     6061 ctacatttcg ctcatcgcca gcccagtcgg gcggcgagtt ccatagcgtt aaggtttcat
     6121 ttagcgcctc aaatagatcc tgttcaggaa ccggatcaaa gagttcctcc gccgctggac
     6181 ctaccaaggc aacgctatgt tctcttgctt ttgtcagcaa gatagccaga tcaatgtcga
     6241 tcgtggctgg ctcgaagata cctgcaagaa tgtcattgcg ctgccattct ccaaattgca
     6301 gttcgcgctt agctggataa cgccacggaa tgatgtcgtc gtgcacaaca atggtgactt
     6361 ctacagcgcg gagaatctcg ctctctccag gggaagccga agtttccaaa aggtcgttga
     6421 tcaaagctcg ccgcgttgtt tcatcaagcc ttacggtcac cgtaaccagc aaatcaatat
     6481 cactgtgtgg cttcaggccg ccatccactg cggagccgta caaatgtacg gccagcaacg
     6541 tcggttcgag atggcgctcg atgacgccaa ctacctctga tagttgagtc gatacttcgg
     6601 cgatcaccgc ttccctcatg atgtttaact cctgaattaa gccgcgccgc gaagcggtgt
     6661 cggcttgaat gaattgttag gcgtcatcct gtgctcccga gaaccagtac cagtacatcg
     6721 ctgtttcgtt cgagacttga ggtctagttt tatacgtgaa caggtcaatg ccgccgagag
     6781 taaagccaca ttttgcgtac aaattgcagg caggtacatt gttcgtttgt gccatgatga
     6841 tatattttta tcttgtgcaa tgtaacatca gagattttga gacacaacgt ggctttgttg
     6901 aataaatcga acttttgctg agttgaagga tcagatcacg catcttcccg acaacgcaga
     6961 ccgttccgtg gcaaagcaaa agttcaaaat caccaactgg tccacctaca acaaagctct
     7021 catcaaccgt ggctccctca ctttctggct ggatgatggg gcgattcagg agatccccat
     7081 ccaacagccc gccgtcgagc gggctttttt atccccggaa gcctgtggat agagggtagt
     7141 tatccacgtg aaaccgctaa tgccccgcaa agccttgatt cacggggctt tccggcccgc
     7201 tccaaaaact atccacgtga aatcgctaat cagggtacgt gaaatcgcta atcggagtac
     7261 gtgaaatcgc taataaggtc acgtgaaatc gctaatcaaa aaggcacgtg agaacgctaa
     7321 tagccctttc agatcaacag cttgcaaaca cccctcgctc cggcaagtag ttacagcaag
     7381 tagtatgttc aattagcttt tcaattatga atatatatat caattattgg tcgcccttgg
     7441 cttgtggaca atgcgctacg cgcaccggct ccgcccgtgg acaaccgcaa gcggttgccc
     7501 accgtcgagc gcctttgccc acaacccggc ggccggccgc aacagatcgt tttataaatt
     7561 tttttttttg aaaaagaaaa agcccgaaag gcggcaacct ctcgggcttc tggatttccg
     7621 atccccggaa ttagactctt ggca
//
