LOCUS       sequence-391165-        6953 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proUBQ10::PIP2A-mCherry-NOSt_1XPhlFO::GFPint-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6953)
  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 6953)
  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: JAB3606
FEATURES             Location/Qualifiers
     source          1..6953
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    8..30
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     CDS             1553..2263
                     /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(1698..1716)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      2282..2534
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        2663..2709
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     primer_bind     complement(2763..2791)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          2931..3119
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     3575..3595
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3966..3986)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3966..3984)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4005..4021)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4005..4021)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4018..4040)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4029..4045
                     /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(4053..4059,4060..4077,4078..4083))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4098..4119
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4236..4253)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4358..4382
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4473..5061)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4553..4572)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5152..5940)
                     /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      6398..6833
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 agactcttgg caggatatat tgtggtgtaa cgttaagctt tggttgtcga cgagtcagta
       61 ataaacggcg tcaaagtggt tgcagccggc acacacgagt cgtgtttatc aactcaaagc
      121 acaaatactt ttcctcaacc taaaaataag gcaattagcc aaaaacaact ttgcgtgtaa
      181 acaacgctca atacacgtgt cattttatta ttagctattg cttcaccgcc ttagctttct
      241 cgtgacctag tcgtcctcgt cttttcttct tcttcttcta taaaacaata cccaaagagc
      301 tcttcttctt cacaattcag atttcaattt ctcaaaatct taaaaacttt ctctcaattc
      361 tctctaccgt gatcaaggta aatttctgtg ttccttattc tctcaaaatc ttcgattttg
      421 ttttcgttcg atcccaattt cgtatatgtt ctttggttta gattctgtta atcttagatc
      481 gtagacgatt ttctgggttt gatcgttaga tatcatctta attctcgatt agggtttcat
      541 agatatcatc cgatttgttc aaataatttg agttttgtcg aataattact cttcgatttg
      601 tgatttctat ctagatctgg tgttagtttc tagtttgtgc gatcgaattt gtcgattaat
      661 ctgagttttt ctgattaaca gcaccatggc aaaggatgtg gaagccgttc ccggagaagg
      721 atttcagaca agagactatc aagatccgcc accagctccg tttattgatg gagcggagct
      781 aaagaagtgg tctttctaca gagcagttat cgcagagttc gtagccactc tcctcttctt
      841 atacatcacc gttttgacag tcatcggtta caagattcag tccgatactg atgccggtgg
      901 cgtagattgc ggcggagttg gaatcctcgg tatcgcttgg gcctttggtg gtatgatctt
      961 catcctcgtc tactgcaccg ccggtatctc tggtggtcac attaacccag cggtgacatt
     1021 tgggctattc ttggcacgta aagtgtcgtt acctagggcc ctattgtaca taatcgctca
     1081 gtgtttgggt gcgatttgtg gagttggttt tgtcaaagcc ttccaaagct cttactacac
     1141 ccgttacgga ggtggagcca actctctagc cgatggctac agcacaggga ccggtctagc
     1201 cgcagagatc attggtactt tcgttcttgt ctacaccgta ttctctgcca ctgaccccaa
     1261 acgtagtgcc agagactccc acgttccggt gttggcgcca cttccaatcg gatttgccgt
     1321 gttcatggta catttggcta ccattcccat taccggaacc ggaattaacc cggcaaggag
     1381 tttcggagct gccgtaatct acaacaagag caagccatgg gatgaccact ggatattttg
     1441 ggttggacca ttcattggag ctgcgatagc tgcattctac caccaattcg ttctaagagc
     1501 ttcaggttct aagtctcttg gatcattcag aagtgctgcc aacgtcggat ccatggtgag
     1561 caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat
     1621 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
     1681 cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg
     1741 ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga
     1801 catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     1861 cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt
     1921 catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa
     1981 taagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa
     2041 gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa
     2101 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa
     2161 gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga
     2221 gggccgccac tccaccggcg gcatggagga gctgtacaag taaagcggcc gcccggctgc
     2281 agatcgttca aacatttggc aataaagttt cttaagattg aatcctgttg ccggtcttgc
     2341 gatgattatc atataatttc tgttgaatta cgttaagcat gtaataatta acatgtaatg
     2401 catgacgtta tttatgagat gggtttttat gattagagtc ccgcaattat acatttaata
     2461 cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg cggtgtcatc
     2521 tatgttacta gatcggggcg cgccctcgag gaagacgtaa tgatagaaaa gttggctgcg
     2581 gccgcgaaat gatacgaaac gtaccgtatc gttaaggtcg ggtctaagct agctaggcca
     2641 cccacaatcc cactatcctt tcgcaagacc cttcctctat ataaggaagt tcatttcatt
     2701 tggagagaac acgggggact ctagacacca tggtgagtaa aggagaagaa cttttcactg
     2761 gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca
     2821 gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta
     2881 ctggaaaact acctgttcca tggccaaccc tggtcaccac cctgacctac gtaagtttct
     2941 gcttctacct ttgatatata tataataatt atcattaatt agtagtaata taatatttca
     3001 aatatttttt tcaaaataaa agaatgtagt atatagcaat tgcttttctg tagtttataa
     3061 gtgtgtatat tttaatttat aacttttcta atatatgacc aaaatttgtt gatgtgcagg
     3121 gcgtgcagtg cttctcccgt taccctgatc atatgaagcg gcacgacttc ttcaagagcg
     3181 ccatgcctga gggatacgtg caggagagga ccatcttctt caaggacgac gggaactaca
     3241 agacacgtgc tgaagtcaag tttgagggtg acaccctcgt caacaggatc gagcttaagg
     3301 gaatcgattt caaggaggac ggaaacatcc tcggccacaa gttggaatac aactacaact
     3361 cccacaacgt atacatcatg gccgacaagc aaaagaacgg catcaaagcc aacttcaaga
     3421 cccgccacaa catcgaggac ggcggcgtgc aactcgctga tcattatcaa caaaatactc
     3481 caattggcga tggccctgtc cttttaccag acaaccatta cctgtccaca caatctgccc
     3541 tttcgaaaga tcccaacgaa aagagagatc acatggtcct tcttgagttt gtaacagctg
     3601 ctgggattac acatggcatg gatgaactat acaaataatg tcaagccatt ctctcgcaga
     3661 tgatgttcac tttgtgtttt acttccttta tgcattcaca gcaataaaag aaagaaatct
     3721 ccatcgcttt tggttttctt ctctgtctta agttagtcgt tttcgtgtct aatctattac
     3781 ttatcattgt aatagactct tcttctattg agatttgaat ataaactaaa acacattcca
     3841 ttttactgtg ttctcaacat tcagaatgca aacggactaa ccgtagtgac tcggcttgag
     3901 tcttcgggct agagcggact agttctagag cggccgccac cgcggtggag ctccagcttt
     3961 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     4021 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     4081 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     4141 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     4201 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4261 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4321 gaatcagggg ataacgcagg aaagaacatg aaggccttga caggatatat tggcgggtaa
     4381 actaagtcgc tgtatgtgtt tgtttgagat ctcatgtgag caaaaggcca gcaaaaggcc
     4441 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4501 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4561 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4621 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4681 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4741 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4801 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4861 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4921 tttggtatct gtgctctgct gaagccagtt accttcggaa gaagagttgg tagctcttga
     4981 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5041 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5101 tggaacgaaa actcacgtta agggattttg gtcatgaatt cttgttagac attatttgcc
     5161 gactaccttg gtgatctcgc ctttcacgta gtgaacaaat tcttccaact gatctgcgcg
     5221 cgaggccaag cgatcttctt gtccaagata agcctgccta gcttcaagta tgacgggctg
     5281 atactgggcc ggcaggcgct ccattgccca gtcggcagcg acatccttcg gcgcgatttt
     5341 gccggttact gcgctgtacc aaatgcggga caacgtaagc actacatttc gctcatcgcc
     5401 agcccagtcg ggcggcgagt tccatagcgt taaggtttca tttagcgcct caaatagatc
     5461 ctgttcagga accggatcaa agagttcctc cgccgctgga cctaccaagg caacgctatg
     5521 ttctcttgct tttgtcagca agatagccag atcaatgtcg atcgtggctg gctcgaagat
     5581 acctgcaaga atgtcattgc gctgccattc tccaaattgc agttcgcgct tagctggata
     5641 acgccacgga atgatgtcgt cgtgcacaac aatggtgact tctacagcgc ggagaatctc
     5701 gctctctcca ggggaagccg aagtttccaa aaggtcgttg atcaaagctc gccgcgttgt
     5761 ttcatcaagc cttacggtca ccgtaaccag caaatcaata tcactgtgtg gcttcaggcc
     5821 gccatccact gcggagccgt acaaatgtac ggccagcaac gtcggttcga gatggcgctc
     5881 gatgacgcca actacctctg atagttgagt cgatacttcg gcgatcaccg cttccctcat
     5941 gatgtttaac tcctgaatta agccgcgccg cgaagcggtg tcggcttgaa tgaattgtta
     6001 ggcgtcatcc tgtgctcccg agaaccagta ccagtacatc gctgtttcgt tcgagacttg
     6061 aggtctagtt ttatacgtga acaggtcaat gccgccgaga gtaaagccac attttgcgta
     6121 caaattgcag gcaggtacat tgttcgtttg tgccatgatg atatattttt atcttgtgca
     6181 atgtaacatc agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct
     6241 gagttgaagg atcagatcac gcatcttccc gacaacgcag accgttccgt ggcaaagcaa
     6301 aagttcaaaa tcaccaactg gtccacctac aacaaagctc tcatcaaccg tggctccctc
     6361 actttctggc tggatgatgg ggcgattcag gagatcccca tccaacagcc cgccgtcgag
     6421 cgggcttttt tatccccgga agcctgtgga tagagggtag ttatccacgt gaaaccgcta
     6481 atgccccgca aagccttgat tcacggggct ttccggcccg ctccaaaaac tatccacgtg
     6541 aaatcgctaa tcagggtacg tgaaatcgct aatcggagta cgtgaaatcg ctaataaggt
     6601 cacgtgaaat cgctaatcaa aaaggcacgt gagaacgcta atagcccttt cagatcaaca
     6661 gcttgcaaac acccctcgct ccggcaagta gttacagcaa gtagtatgtt caattagctt
     6721 ttcaattatg aatatatata tcaattattg gtcgcccttg gcttgtggac aatgcgctac
     6781 gcgcaccggc tccgcccgtg gacaaccgca agcggttgcc caccgtcgag cgcctttgcc
     6841 cacaacccgg cggccggccg caacagatcg ttttataaat tttttttttt gaaaaagaaa
     6901 aagcccgaaa ggcggcaacc tctcgggctt ctggatttcc gatccccgga att
//
