LOCUS       sequence-389451-        7707 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants 
            (proUBQ10::PIP2A-mCherry-NOSt_pro35S-IcaRO::GFPint-NLS-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7707)
  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 7707)
  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: JAB2461
FEATURES             Location/Qualifiers
     source          1..7707
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    44..66
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     CDS             1589..2299
                     /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(1734..1752)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      2318..2570
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        3093..3438
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     3381..3401
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     complement(3523..3551)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          3691..3879
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     4335..4355
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             4396..4416
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4756..4776)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4756..4774)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4795..4811)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4795..4811)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4808..4830)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4819..4835
                     /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(4843..4849,4850..4867,4868..4873))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4888..4909
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5026..5043)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5148..5172
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5263..5851)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5343..5362)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5942..6730)
                     /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      7188..7623
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 acctctcggg cttctggatt tccgatcccc ggaattagac tcttggcagg atatattgtg
       61 gtgtaacgtt aagctttggt tgtcgacgag tcagtaataa acggcgtcaa agtggttgca
      121 gccggcacac acgagtcgtg tttatcaact caaagcacaa atacttttcc tcaacctaaa
      181 aataaggcaa ttagccaaaa acaactttgc gtgtaaacaa cgctcaatac acgtgtcatt
      241 ttattattag ctattgcttc accgccttag ctttctcgtg acctagtcgt cctcgtcttt
      301 tcttcttctt cttctataaa acaataccca aagagctctt cttcttcaca attcagattt
      361 caatttctca aaatcttaaa aactttctct caattctctc taccgtgatc aaggtaaatt
      421 tctgtgttcc ttattctctc aaaatcttcg attttgtttt cgttcgatcc caatttcgta
      481 tatgttcttt ggtttagatt ctgttaatct tagatcgtag acgattttct gggtttgatc
      541 gttagatatc atcttaattc tcgattaggg tttcatagat atcatccgat ttgttcaaat
      601 aatttgagtt ttgtcgaata attactcttc gatttgtgat ttctatctag atctggtgtt
      661 agtttctagt ttgtgcgatc gaatttgtcg attaatctga gtttttctga ttaacagcac
      721 catggcaaag gatgtggaag ccgttcccgg agaaggattt cagacaagag actatcaaga
      781 tccgccacca gctccgttta ttgatggagc ggagctaaag aagtggtctt tctacagagc
      841 agttatcgca gagttcgtag ccactctcct cttcttatac atcaccgttt tgacagtcat
      901 cggttacaag attcagtccg atactgatgc cggtggcgta gattgcggcg gagttggaat
      961 cctcggtatc gcttgggcct ttggtggtat gatcttcatc ctcgtctact gcaccgccgg
     1021 tatctctggt ggtcacatta acccagcggt gacatttggg ctattcttgg cacgtaaagt
     1081 gtcgttacct agggccctat tgtacataat cgctcagtgt ttgggtgcga tttgtggagt
     1141 tggttttgtc aaagccttcc aaagctctta ctacacccgt tacggaggtg gagccaactc
     1201 tctagccgat ggctacagca cagggaccgg tctagccgca gagatcattg gtactttcgt
     1261 tcttgtctac accgtattct ctgccactga ccccaaacgt agtgccagag actcccacgt
     1321 tccggtgttg gcgccacttc caatcggatt tgccgtgttc atggtacatt tggctaccat
     1381 tcccattacc ggaaccggaa ttaacccggc aaggagtttc ggagctgccg taatctacaa
     1441 caagagcaag ccatgggatg accactggat attttgggtt ggaccattca ttggagctgc
     1501 gatagctgca ttctaccacc aattcgttct aagagcttca ggttctaagt ctcttggatc
     1561 attcagaagt gctgccaacg tcggatccat ggtgagcaag ggcgaggagg ataacatggc
     1621 catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga
     1681 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     1741 gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat
     1801 gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc
     1861 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
     1921 cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg
     1981 caccaacttc ccctccgacg gccccgtaat gcagaataag accatgggct gggaggcctc
     2041 ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa
     2101 gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc
     2161 cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga
     2221 ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat
     2281 ggaggagctg tacaagtaaa gcggccgccc ggctgcagat cgttcaaaca tttggcaata
     2341 aagtttctta agattgaatc ctgttgccgg tcttgcgatg attatcatat aatttctgtt
     2401 gaattacgtt aagcatgtaa taattaacat gtaatgcatg acgttattta tgagatgggt
     2461 ttttatgatt agagtcccgc aattatacat ttaatacgcg atagaaaaca aaatatagcg
     2521 cgcaaactag gataaattat cgcgcgcggt gtcatctatg ttactagatc ggggcgcgcc
     2581 ctcgaggaag acgtaatggg tccccagatt tgccttttca atttcagaaa gaatgctaac
     2641 ccacagatgg ttagagaggc ttacgcagca gctctcatca agacgatcta cccgagcaat
     2701 aatctccagg aaatcaaata ccttcccaag aaggttaaag atgcagtcaa aagattcagg
     2761 actaactgca tcaagaacac agagaaagat atatttctca agatcagaag tactattcca
     2821 gtatggacga ttcaaggctt gcttcacaaa ccaaggcaag taatagagat tggagtctct
     2881 aaaaaggtag ttcccactga atcaaaggcc atggagtcaa agattcaaat agaggaccta
     2941 acagaactcg ccgtaaagac tggcgaacag ttcatacaga gtctcttacg actcaatgac
     3001 aagaagaaaa tcttcgtcaa catggtggag cacgacacac ttgtctactc caaaaatatc
     3061 aaagatacag tctcagaaga ccaaagggca attgagactt ttcaacaaag ggtaatatcc
     3121 ggaaacctcc tcggattcca ttgcccagct atctgtcact ttattgtgaa gatagtggaa
     3181 aaggaaggtg gctcctacaa atgccatcat tgcgataaag gaaaggccat cgttgaagat
     3241 gcctctgccg acagtggtcc caaagatgga cccccaccca cgaggagcat cgtggaaaaa
     3301 gaagacgttc caaccacgtc ttcaaagcaa gtggattgat gtgatatctc cactgacgta
     3361 agggatgacg cacaatccca ctatccttcg caagaccctt cctctatata aggaagttca
     3421 tttcatttgg agagaacacg ggggactcta gattgattca cctacctttc gttaggttag
     3481 gttgtcacca tggtgagtaa aggagaagaa cttttcactg gagttgtccc aattcttgtt
     3541 gaattagatg gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat
     3601 gcaacatacg gaaaacttac ccttaaattt atttgcacta ctggaaaact acctgttcca
     3661 tggccaaccc tggtcaccac cctgacctac gtaagtttct gcttctacct ttgatatata
     3721 tataataatt atcattaatt agtagtaata taatatttca aatatttttt tcaaaataaa
     3781 agaatgtagt atatagcaat tgcttttctg tagtttataa gtgtgtatat tttaatttat
     3841 aacttttcta atatatgacc aaaatttgtt gatgtgcagg gcgtgcagtg cttctcccgt
     3901 taccctgatc atatgaagcg gcacgacttc ttcaagagcg ccatgcctga gggatacgtg
     3961 caggagagga ccatcttctt caaggacgac gggaactaca agacacgtgc tgaagtcaag
     4021 tttgagggtg acaccctcgt caacaggatc gagcttaagg gaatcgattt caaggaggac
     4081 ggaaacatcc tcggccacaa gttggaatac aactacaact cccacaacgt atacatcatg
     4141 gccgacaagc aaaagaacgg catcaaagcc aacttcaaga cccgccacaa catcgaggac
     4201 ggcggcgtgc aactcgctga tcattatcaa caaaatactc caattggcga tggccctgtc
     4261 cttttaccag acaaccatta cctgtccaca caatctgccc tttcgaaaga tcccaacgaa
     4321 aagagagatc acatggtcct tcttgagttt gtaacagctg ctgggattac acatggcatg
     4381 gatgaactat acaaacctaa gaaaaagcgt aaggtcgagg acccttaatg tcaagccatt
     4441 ctctcgcaga tgatgttcac tttgtgtttt acttccttta tgcattcaca gcaataaaag
     4501 aaagaaatct ccatcgcttt tggttttctt ctctgtctta agttagtcgt tttcgtgtct
     4561 aatctattac ttatcattgt aatagactct tcttctattg agatttgaat ataaactaaa
     4621 acacattcca ttttactgtg ttctcaacat tcagaatgca aacggactaa ccgtagtgac
     4681 tcggcttgag tcttcgggct agagcggact agttctagag cggccgccac cgcggtggag
     4741 ctccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
     4801 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4861 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4921 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4981 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5041 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5101 gttatccaca gaatcagggg ataacgcagg aaagaacatg aaggccttga caggatatat
     5161 tggcgggtaa actaagtcgc tgtatgtgtt tgtttgagat ctcatgtgag caaaaggcca
     5221 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5281 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5341 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5401 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5461 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5521 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5581 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5641 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5701 aagaacagta tttggtatct gtgctctgct gaagccagtt accttcggaa gaagagttgg
     5761 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5821 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5881 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgaatt cttgttagac
     5941 attatttgcc gactaccttg gtgatctcgc ctttcacgta gtgaacaaat tcttccaact
     6001 gatctgcgcg cgaggccaag cgatcttctt gtccaagata agcctgccta gcttcaagta
     6061 tgacgggctg atactgggcc ggcaggcgct ccattgccca gtcggcagcg acatccttcg
     6121 gcgcgatttt gccggttact gcgctgtacc aaatgcggga caacgtaagc actacatttc
     6181 gctcatcgcc agcccagtcg ggcggcgagt tccatagcgt taaggtttca tttagcgcct
     6241 caaatagatc ctgttcagga accggatcaa agagttcctc cgccgctgga cctaccaagg
     6301 caacgctatg ttctcttgct tttgtcagca agatagccag atcaatgtcg atcgtggctg
     6361 gctcgaagat acctgcaaga atgtcattgc gctgccattc tccaaattgc agttcgcgct
     6421 tagctggata acgccacgga atgatgtcgt cgtgcacaac aatggtgact tctacagcgc
     6481 ggagaatctc gctctctcca ggggaagccg aagtttccaa aaggtcgttg atcaaagctc
     6541 gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag caaatcaata tcactgtgtg
     6601 gcttcaggcc gccatccact gcggagccgt acaaatgtac ggccagcaac gtcggttcga
     6661 gatggcgctc gatgacgcca actacctctg atagttgagt cgatacttcg gcgatcaccg
     6721 cttccctcat gatgtttaac tcctgaatta agccgcgccg cgaagcggtg tcggcttgaa
     6781 tgaattgtta ggcgtcatcc tgtgctcccg agaaccagta ccagtacatc gctgtttcgt
     6841 tcgagacttg aggtctagtt ttatacgtga acaggtcaat gccgccgaga gtaaagccac
     6901 attttgcgta caaattgcag gcaggtacat tgttcgtttg tgccatgatg atatattttt
     6961 atcttgtgca atgtaacatc agagattttg agacacaacg tggctttgtt gaataaatcg
     7021 aacttttgct gagttgaagg atcagatcac gcatcttccc gacaacgcag accgttccgt
     7081 ggcaaagcaa aagttcaaaa tcaccaactg gtccacctac aacaaagctc tcatcaaccg
     7141 tggctccctc actttctggc tggatgatgg ggcgattcag gagatcccca tccaacagcc
     7201 cgccgtcgag cgggcttttt tatccccgga agcctgtgga tagagggtag ttatccacgt
     7261 gaaaccgcta atgccccgca aagccttgat tcacggggct ttccggcccg ctccaaaaac
     7321 tatccacgtg aaatcgctaa tcagggtacg tgaaatcgct aatcggagta cgtgaaatcg
     7381 ctaataaggt cacgtgaaat cgctaatcaa aaaggcacgt gagaacgcta atagcccttt
     7441 cagatcaaca gcttgcaaac acccctcgct ccggcaagta gttacagcaa gtagtatgtt
     7501 caattagctt ttcaattatg aatatatata tcaattattg gtcgcccttg gcttgtggac
     7561 aatgcgctac gcgcaccggc tccgcccgtg gacaaccgca agcggttgcc caccgtcgag
     7621 cgcctttgcc cacaacccgg cggccggccg caacagatcg ttttataaat tttttttttt
     7681 gaaaaagaaa aagcccgaaa ggcggca
//
