LOCUS       sequence-389440-        7033 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants 
            (proUBQ10::PIP2A-mCherry-NOSt_proG1090::GFPint-ADHterm).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7033)
  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 7033)
  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: JAB2224
FEATURES             Location/Qualifiers
     source          1..7033
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     191..219
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     promoter        complement(271..317)
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     primer_bind     complement(306..326)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(528..780)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(799..1509)
                     /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     1346..1364
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     misc_feature    3032..3054
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     rep_origin      3182..3617
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     CDS             4075..4863
                     /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      4954..5542
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5443..5462
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5633..5657
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     5762..5779
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5896..5917
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5932..5937,5938..5955,5956..5962)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5970..5986
                     /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)."
     primer_bind     5975..5997
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5994..6010
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5994..6010
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6029..6049
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6031..6049
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6420..6440)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     intron          join(6896..7033,1..51)
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
ORIGIN
        1 taattaatga taattattat atatatatca aaggtagaag cagaaactta cgtaggtcag
       61 ggtggtgacc agggttggcc atggaacagg tagttttcca gtagtgcaaa taaatttaag
      121 ggtaagtttt ccgtatgttg catcaccttc accctctcca ctgacagaaa atttgtgccc
      181 attaacatca ccatctaatt caacaagaat tgggacaact ccagtgaaaa gttcttctcc
      241 tttactcacc atggtgggga tcccagcgtg tcctctccaa atgaaatgaa cttccttata
      301 tagaggaagg gtcttgcgaa ggatagtggg attgtgcgtc atcccttacg tcagtggaga
      361 tggatcctct agaggcacgt ggcggcacgt ggcggcacgt ggcggcacgt ggcaagcttg
      421 gatccacgat atcctgcagg catgcaagct tagcttgagc ttggatcaga ttgtcgtttc
      481 ccgccttcag tttaaactac attacgtctt cctcgagggc gcgccccgat ctagtaacat
      541 agatgacacc gcgcgcgata atttatccta gtttgcgcgc tatattttgt tttctatcgc
      601 gtattaaatg tataattgcg ggactctaat cataaaaacc catctcataa ataacgtcat
      661 gcattacatg ttaattatta catgcttaac gtaattcaac agaaattata tgataatcat
      721 cgcaagaccg gcaacaggat tcaatcttaa gaaactttat tgccaaatgt ttgaacgatc
      781 tgcagccggg cggccgcttt acttgtacag ctcctccatg ccgccggtgg agtggcggcc
      841 ctcggcgcgt tcgtactgtt ccacgatggt gtagtcctcg ttgtgggagg tgatgtccaa
      901 cttgatgttg acgttgtagg cgccgggcag ctgcacgggc ttcttggcct tgtaggtggt
      961 cttgacctca gcgtcgtagt ggccgccgtc cttcagcttc agcctctgct tgatctcgcc
     1021 cttcagggcg ccgtcctcgg ggtacatccg ctcggaggag gcctcccagc ccatggtctt
     1081 attctgcatt acggggccgt cggaggggaa gttggtgccg cgcagcttca ccttgtagat
     1141 gaactcgccg tcctgcaggg aggagtcctg ggtcacggtc accacgccgc cgtcctcgaa
     1201 gttcatcacg cgctcccact tgaagccctc ggggaaggac agcttcaagt agtcggggat
     1261 gtcggcgggg tgcttcacgt aggccttgga gccgtacatg aactgagggg acaggatgtc
     1321 ccaggcgaag ggcagggggc cacccttggt caccttcagc ttggcggtct gggtgccctc
     1381 gtaggggcgg ccctcgccct cgccctcgat ctcgaactcg tggccgttca cggagccctc
     1441 catgtgcacc ttgaagcgca tgaactcctt gatgatggcc atgttatcct cctcgccctt
     1501 gctcaccatg gatccgacgt tggcagcact tctgaatgat ccaagagact tagaacctga
     1561 agctcttaga acgaattggt ggtagaatgc agctatcgca gctccaatga atggtccaac
     1621 ccaaaatatc cagtggtcat cccatggctt gctcttgttg tagattacgg cagctccgaa
     1681 actccttgcc gggttaattc cggttccggt aatgggaatg gtagccaaat gtaccatgaa
     1741 cacggcaaat ccgattggaa gtggcgccaa caccggaacg tgggagtctc tggcactacg
     1801 tttggggtca gtggcagaga atacggtgta gacaagaacg aaagtaccaa tgatctctgc
     1861 ggctagaccg gtccctgtgc tgtagccatc ggctagagag ttggctccac ctccgtaacg
     1921 ggtgtagtaa gagctttgga aggctttgac aaaaccaact ccacaaatcg cacccaaaca
     1981 ctgagcgatt atgtacaata gggccctagg taacgacact ttacgtgcca agaatagccc
     2041 aaatgtcacc gctgggttaa tgtgaccacc agagataccg gcggtgcagt agacgaggat
     2101 gaagatcata ccaccaaagg cccaagcgat accgaggatt ccaactccgc cgcaatctac
     2161 gccaccggca tcagtatcgg actgaatctt gtaaccgatg actgtcaaaa cggtgatgta
     2221 taagaagagg agagtggcta cgaactctgc gataactgct ctgtagaaag accacttctt
     2281 tagctccgct ccatcaataa acggagctgg tggcggatct tgatagtctc ttgtctgaaa
     2341 tccttctccg ggaacggctt ccacatcctt tgccatggtg ctgttaatca gaaaaactca
     2401 gattaatcga caaattcgat cgcacaaact agaaactaac accagatcta gatagaaatc
     2461 acaaatcgaa gagtaattat tcgacaaaac tcaaattatt tgaacaaatc ggatgatatc
     2521 tatgaaaccc taatcgagaa ttaagatgat atctaacgat caaacccaga aaatcgtcta
     2581 cgatctaaga ttaacagaat ctaaaccaaa gaacatatac gaaattggga tcgaacgaaa
     2641 acaaaatcga agattttgag agaataagga acacagaaat ttaccttgat cacggtagag
     2701 agaattgaga gaaagttttt aagattttga gaaattgaaa tctgaattgt gaagaagaag
     2761 agctctttgg gtattgtttt atagaagaag aagaagaaaa gacgaggacg actaggtcac
     2821 gagaaagcta aggcggtgaa gcaatagcta ataataaaat gacacgtgta ttgagcgttg
     2881 tttacacgca aagttgtttt tggctaattg ccttattttt aggttgagga aaagtatttg
     2941 tgctttgagt tgataaacac gactcgtgtg tgccggctgc aaccactttg acgccgttta
     3001 ttactgactc gtcgacaacc aaagcttaac gttacaccac aatatatcct gccaagagtc
     3061 taattccggg gatcggaaat ccagaagccc gagaggttgc cgcctttcgg gctttttctt
     3121 tttcaaaaaa aaaaatttat aaaacgatct gttgcggccg gccgccgggt tgtgggcaaa
     3181 ggcgctcgac ggtgggcaac cgcttgcggt tgtccacggg cggagccggt gcgcgtagcg
     3241 cattgtccac aagccaaggg cgaccaataa ttgatatata tattcataat tgaaaagcta
     3301 attgaacata ctacttgctg taactacttg ccggagcgag gggtgtttgc aagctgttga
     3361 tctgaaaggg ctattagcgt tctcacgtgc ctttttgatt agcgatttca cgtgacctta
     3421 ttagcgattt cacgtactcc gattagcgat ttcacgtacc ctgattagcg atttcacgtg
     3481 gatagttttt ggagcgggcc ggaaagcccc gtgaatcaag gctttgcggg gcattagcgg
     3541 tttcacgtgg ataactaccc tctatccaca ggcttccggg gataaaaaag cccgctcgac
     3601 ggcgggctgt tggatgggga tctcctgaat cgccccatca tccagccaga aagtgaggga
     3661 gccacggttg atgagagctt tgttgtaggt ggaccagttg gtgattttga acttttgctt
     3721 tgccacggaa cggtctgcgt tgtcgggaag atgcgtgatc tgatccttca actcagcaaa
     3781 agttcgattt attcaacaaa gccacgttgt gtctcaaaat ctctgatgtt acattgcaca
     3841 agataaaaat atatcatcat ggcacaaacg aacaatgtac ctgcctgcaa tttgtacgca
     3901 aaatgtggct ttactctcgg cggcattgac ctgttcacgt ataaaactag acctcaagtc
     3961 tcgaacgaaa cagcgatgta ctggtactgg ttctcgggag cacaggatga cgcctaacaa
     4021 ttcattcaag ccgacaccgc ttcgcggcgc ggcttaattc aggagttaaa catcatgagg
     4081 gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc
     4141 catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg
     4201 aagccacaca gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg
     4261 cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt
     4321 ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca
     4381 gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc
     4441 gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc
     4501 gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta
     4561 tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat
     4621 gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc
     4681 gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc
     4741 gtcatacttg aagctaggca ggcttatctt ggacaagaag atcgcttggc ctcgcgcgca
     4801 gatcagttgg aagaatttgt tcactacgtg aaaggcgaga tcaccaaggt agtcggcaaa
     4861 taatgtctaa caagaattca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4921 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4981 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5041 gctaccaact cttcttccga aggtaactgg cttcagcaga gcacagatac caaatactgt
     5101 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5161 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5221 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5281 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5341 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5401 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5461 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5521 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5581 ttgctggcct tttgctcaca tgagatctca aacaaacaca tacagcgact tagtttaccc
     5641 gccaatatat cctgtcaagg ccttcatgtt ctttcctgcg ttatcccctg attctgtgga
     5701 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     5761 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     5821 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     5881 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     5941 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     6001 cagctatgac catgattacg ccaagcgcgc aattaaccct cactaaaggg aacaaaagct
     6061 ggagctccac cgcggtggcg gccgctctag aactagtccg ctctagcccg aagactcaag
     6121 ccgagtcact acggttagtc cgtttgcatt ctgaatgttg agaacacagt aaaatggaat
     6181 gtgttttagt ttatattcaa atctcaatag aagaagagtc tattacaatg ataagtaata
     6241 gattagacac gaaaacgact aacttaagac agagaagaaa accaaaagcg atggagattt
     6301 ctttctttta ttgctgtgaa tgcataaagg aagtaaaaca caaagtgaac atcatctgcg
     6361 agagaatggc ttgacattat ttgtatagtt catccatgcc atgtgtaatc ccagcagctg
     6421 ttacaaactc aagaaggacc atgtgatctc tcttttcgtt gggatctttc gaaagggcag
     6481 attgtgtgga caggtaatgg ttgtctggta aaaggacagg gccatcgcca attggagtat
     6541 tttgttgata atgatcagcg agttgcacgc cgccgtcttc gatgttgtgg cgggtcttga
     6601 agttggcttt gatgccgttc ttttgcttgt cggccatgat gtatacgttg tgggagttgt
     6661 agttgtattc caacttgtgg ccgaggatgt ttccgtcctc cttgaaatcg attcccttaa
     6721 gctcgatcct gttgacgagg gtgtctccct caaacttgac ttcagcacgt gtcttgtagt
     6781 tcccgtcgtc cttgaagaag atggtcctct cctgcacgta tccctcaggc atggcgctct
     6841 tgaagaagtc gtgccgcttc atatgatcag ggtaacggga gaagcactgc acgccctgca
     6901 catcaacaaa ttttggtcat atattagaaa agttataaat taaaatatac acacttataa
     6961 actacagaaa agcaattgct atatactaca ttcttttatt ttgaaaaaaa tatttgaaat
     7021 attatattac tac
//
