LOCUS       sequence-389458-        7702 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants 
            (proUBQ10::PIP2A-mCherry-NOSt_pro35S-LmrAO::GFPint-NLS-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7702)
  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 7702)
  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: JAB2462
FEATURES             Location/Qualifiers
     source          1..7702
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    26..48
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     CDS             1571..2281
                     /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(1716..1734)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      2300..2552
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        3075..3420
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     3363..3383
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     complement(3500..3528)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          3668..3856
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     4312..4332
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             4373..4393
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4733..4753)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4733..4751)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4772..4788)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4772..4788)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4785..4807)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4796..4812
                     /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(4820..4826,4827..4844,4845..4850))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4865..4886
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5003..5020)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5125..5149
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5240..5828)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5320..5339)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5919..6707)
                     /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      7165..7600
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 tttccgatcc ccggaattag actcttggca ggatatattg tggtgtaacg ttaagctttg
       61 gttgtcgacg agtcagtaat aaacggcgtc aaagtggttg cagccggcac acacgagtcg
      121 tgtttatcaa ctcaaagcac aaatactttt cctcaaccta aaaataaggc aattagccaa
      181 aaacaacttt gcgtgtaaac aacgctcaat acacgtgtca ttttattatt agctattgct
      241 tcaccgcctt agctttctcg tgacctagtc gtcctcgtct tttcttcttc ttcttctata
      301 aaacaatacc caaagagctc ttcttcttca caattcagat ttcaatttct caaaatctta
      361 aaaactttct ctcaattctc tctaccgtga tcaaggtaaa tttctgtgtt ccttattctc
      421 tcaaaatctt cgattttgtt ttcgttcgat cccaatttcg tatatgttct ttggtttaga
      481 ttctgttaat cttagatcgt agacgatttt ctgggtttga tcgttagata tcatcttaat
      541 tctcgattag ggtttcatag atatcatccg atttgttcaa ataatttgag ttttgtcgaa
      601 taattactct tcgatttgtg atttctatct agatctggtg ttagtttcta gtttgtgcga
      661 tcgaatttgt cgattaatct gagtttttct gattaacagc accatggcaa aggatgtgga
      721 agccgttccc ggagaaggat ttcagacaag agactatcaa gatccgccac cagctccgtt
      781 tattgatgga gcggagctaa agaagtggtc tttctacaga gcagttatcg cagagttcgt
      841 agccactctc ctcttcttat acatcaccgt tttgacagtc atcggttaca agattcagtc
      901 cgatactgat gccggtggcg tagattgcgg cggagttgga atcctcggta tcgcttgggc
      961 ctttggtggt atgatcttca tcctcgtcta ctgcaccgcc ggtatctctg gtggtcacat
     1021 taacccagcg gtgacatttg ggctattctt ggcacgtaaa gtgtcgttac ctagggccct
     1081 attgtacata atcgctcagt gtttgggtgc gatttgtgga gttggttttg tcaaagcctt
     1141 ccaaagctct tactacaccc gttacggagg tggagccaac tctctagccg atggctacag
     1201 cacagggacc ggtctagccg cagagatcat tggtactttc gttcttgtct acaccgtatt
     1261 ctctgccact gaccccaaac gtagtgccag agactcccac gttccggtgt tggcgccact
     1321 tccaatcgga tttgccgtgt tcatggtaca tttggctacc attcccatta ccggaaccgg
     1381 aattaacccg gcaaggagtt tcggagctgc cgtaatctac aacaagagca agccatggga
     1441 tgaccactgg atattttggg ttggaccatt cattggagct gcgatagctg cattctacca
     1501 ccaattcgtt ctaagagctt caggttctaa gtctcttgga tcattcagaa gtgctgccaa
     1561 cgtcggatcc atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat
     1621 gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga
     1681 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg
     1741 ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta
     1801 cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa
     1861 gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc
     1921 cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga
     1981 cggccccgta atgcagaata agaccatggg ctgggaggcc tcctccgagc ggatgtaccc
     2041 cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca
     2101 ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc
     2161 ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga
     2221 acagtacgaa cgcgccgagg gccgccactc caccggcggc atggaggagc tgtacaagta
     2281 aagcggccgc ccggctgcag atcgttcaaa catttggcaa taaagtttct taagattgaa
     2341 tcctgttgcc ggtcttgcga tgattatcat ataatttctg ttgaattacg ttaagcatgt
     2401 aataattaac atgtaatgca tgacgttatt tatgagatgg gtttttatga ttagagtccc
     2461 gcaattatac atttaatacg cgatagaaaa caaaatatag cgcgcaaact aggataaatt
     2521 atcgcgcgcg gtgtcatcta tgttactaga tcggggcgcg ccctcgagga agacgtaatg
     2581 ggtccccaga tttgcctttt caatttcaga aagaatgcta acccacagat ggttagagag
     2641 gcttacgcag cagctctcat caagacgatc tacccgagca ataatctcca ggaaatcaaa
     2701 taccttccca agaaggttaa agatgcagtc aaaagattca ggactaactg catcaagaac
     2761 acagagaaag atatatttct caagatcaga agtactattc cagtatggac gattcaaggc
     2821 ttgcttcaca aaccaaggca agtaatagag attggagtct ctaaaaaggt agttcccact
     2881 gaatcaaagg ccatggagtc aaagattcaa atagaggacc taacagaact cgccgtaaag
     2941 actggcgaac agttcataca gagtctctta cgactcaatg acaagaagaa aatcttcgtc
     3001 aacatggtgg agcacgacac acttgtctac tccaaaaata tcaaagatac agtctcagaa
     3061 gaccaaaggg caattgagac ttttcaacaa agggtaatat ccggaaacct cctcggattc
     3121 cattgcccag ctatctgtca ctttattgtg aagatagtgg aaaaggaagg tggctcctac
     3181 aaatgccatc attgcgataa aggaaaggcc atcgttgaag atgcctctgc cgacagtggt
     3241 cccaaagatg gacccccacc cacgaggagc atcgtggaaa aagaagacgt tccaaccacg
     3301 tcttcaaagc aagtggattg atgtgatatc tccactgacg taagggatga cgcacaatcc
     3361 cactatcctt cgcaagaccc ttcctctata taaggaagtt catttcattt ggagagaaca
     3421 cgggggactc tagattgaga taatagacca gtcactatat ttcaccatgg tgagtaaagg
     3481 agaagaactt ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg
     3541 gcacaaattt tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct
     3601 taaatttatt tgcactactg gaaaactacc tgttccatgg ccaaccctgg tcaccaccct
     3661 gacctacgta agtttctgct tctacctttg atatatatat aataattatc attaattagt
     3721 agtaatataa tatttcaaat atttttttca aaataaaaga atgtagtata tagcaattgc
     3781 ttttctgtag tttataagtg tgtatatttt aatttataac ttttctaata tatgaccaaa
     3841 atttgttgat gtgcagggcg tgcagtgctt ctcccgttac cctgatcata tgaagcggca
     3901 cgacttcttc aagagcgcca tgcctgaggg atacgtgcag gagaggacca tcttcttcaa
     3961 ggacgacggg aactacaaga cacgtgctga agtcaagttt gagggtgaca ccctcgtcaa
     4021 caggatcgag cttaagggaa tcgatttcaa ggaggacgga aacatcctcg gccacaagtt
     4081 ggaatacaac tacaactccc acaacgtata catcatggcc gacaagcaaa agaacggcat
     4141 caaagccaac ttcaagaccc gccacaacat cgaggacggc ggcgtgcaac tcgctgatca
     4201 ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct
     4261 gtccacacaa tctgcccttt cgaaagatcc caacgaaaag agagatcaca tggtccttct
     4321 tgagtttgta acagctgctg ggattacaca tggcatggat gaactataca aacctaagaa
     4381 aaagcgtaag gtcgaggacc cttaatgtca agccattctc tcgcagatga tgttcacttt
     4441 gtgttttact tcctttatgc attcacagca ataaaagaaa gaaatctcca tcgcttttgg
     4501 ttttcttctc tgtcttaagt tagtcgtttt cgtgtctaat ctattactta tcattgtaat
     4561 agactcttct tctattgaga tttgaatata aactaaaaca cattccattt tactgtgttc
     4621 tcaacattca gaatgcaaac ggactaaccg tagtgactcg gcttgagtct tcgggctaga
     4681 gcggactagt tctagagcgg ccgccaccgc ggtggagctc cagcttttgt tccctttagt
     4741 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4801 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4861 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4921 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4981 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5041 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5101 acgcaggaaa gaacatgaag gccttgacag gatatattgg cgggtaaact aagtcgctgt
     5161 atgtgtttgt ttgagatctc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5221 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5281 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5341 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5401 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5461 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5521 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5581 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5641 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgtg
     5701 ctctgctgaa gccagttacc ttcggaagaa gagttggtag ctcttgatcc ggcaaacaaa
     5761 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5821 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5881 cacgttaagg gattttggtc atgaattctt gttagacatt atttgccgac taccttggtg
     5941 atctcgcctt tcacgtagtg aacaaattct tccaactgat ctgcgcgcga ggccaagcga
     6001 tcttcttgtc caagataagc ctgcctagct tcaagtatga cgggctgata ctgggccggc
     6061 aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc ggttactgcg
     6121 ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct catcgccagc ccagtcgggc
     6181 ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa atagatcctg ttcaggaacc
     6241 ggatcaaaga gttcctccgc cgctggacct accaaggcaa cgctatgttc tcttgctttt
     6301 gtcagcaaga tagccagatc aatgtcgatc gtggctggct cgaagatacc tgcaagaatg
     6361 tcattgcgct gccattctcc aaattgcagt tcgcgcttag ctggataacg ccacggaatg
     6421 atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga gaatctcgct ctctccaggg
     6481 gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc atcaagcctt
     6541 acggtcaccg taaccagcaa atcaatatca ctgtgtggct tcaggccgcc atccactgcg
     6601 gagccgtaca aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat gacgccaact
     6661 acctctgata gttgagtcga tacttcggcg atcaccgctt ccctcatgat gtttaactcc
     6721 tgaattaagc cgcgccgcga agcggtgtcg gcttgaatga attgttaggc gtcatcctgt
     6781 gctcccgaga accagtacca gtacatcgct gtttcgttcg agacttgagg tctagtttta
     6841 tacgtgaaca ggtcaatgcc gccgagagta aagccacatt ttgcgtacaa attgcaggca
     6901 ggtacattgt tcgtttgtgc catgatgata tatttttatc ttgtgcaatg taacatcaga
     6961 gattttgaga cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc
     7021 agatcacgca tcttcccgac aacgcagacc gttccgtggc aaagcaaaag ttcaaaatca
     7081 ccaactggtc cacctacaac aaagctctca tcaaccgtgg ctccctcact ttctggctgg
     7141 atgatggggc gattcaggag atccccatcc aacagcccgc cgtcgagcgg gcttttttat
     7201 ccccggaagc ctgtggatag agggtagtta tccacgtgaa accgctaatg ccccgcaaag
     7261 ccttgattca cggggctttc cggcccgctc caaaaactat ccacgtgaaa tcgctaatca
     7321 gggtacgtga aatcgctaat cggagtacgt gaaatcgcta ataaggtcac gtgaaatcgc
     7381 taatcaaaaa ggcacgtgag aacgctaata gccctttcag atcaacagct tgcaaacacc
     7441 cctcgctccg gcaagtagtt acagcaagta gtatgttcaa ttagcttttc aattatgaat
     7501 atatatatca attattggtc gcccttggct tgtggacaat gcgctacgcg caccggctcc
     7561 gcccgtggac aaccgcaagc ggttgcccac cgtcgagcgc ctttgcccac aacccggcgg
     7621 ccggccgcaa cagatcgttt tataaatttt tttttttgaa aaagaaaaag cccgaaaggc
     7681 ggcaacctct cgggcttctg ga
//
