LOCUS       sequence-389460-        7695 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants 
            (proUBQ10::PIP2A-mCherry-NOSt_pro35S-McbRO::GFPint-NLS-ADHt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7695)
  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 7695)
  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: JAB2485
FEATURES             Location/Qualifiers
     source          1..7695
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    46..68
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     CDS             1591..2301
                     /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(1736..1754)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      2320..2572
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        3095..3440
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     3383..3403
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     complement(3513..3541)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          3681..3869
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     4325..4345
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             4386..4406
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4746..4766)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4746..4764)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4785..4801)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4785..4801)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4798..4820)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4809..4825
                     /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(4833..4839,4840..4857,4858..4863))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4878..4899
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5016..5033)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5138..5162
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5253..5841)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5333..5352)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5932..6720)
                     /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      7178..7613
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 caacctctcg ggcttctgga tttccgatcc ccggaattag actcttggca ggatatattg
       61 tggtgtaacg ttaagctttg gttgtcgacg agtcagtaat aaacggcgtc aaagtggttg
      121 cagccggcac acacgagtcg tgtttatcaa ctcaaagcac aaatactttt cctcaaccta
      181 aaaataaggc aattagccaa aaacaacttt gcgtgtaaac aacgctcaat acacgtgtca
      241 ttttattatt agctattgct tcaccgcctt agctttctcg tgacctagtc gtcctcgtct
      301 tttcttcttc ttcttctata aaacaatacc caaagagctc ttcttcttca caattcagat
      361 ttcaatttct caaaatctta aaaactttct ctcaattctc tctaccgtga tcaaggtaaa
      421 tttctgtgtt ccttattctc tcaaaatctt cgattttgtt ttcgttcgat cccaatttcg
      481 tatatgttct ttggtttaga ttctgttaat cttagatcgt agacgatttt ctgggtttga
      541 tcgttagata tcatcttaat tctcgattag ggtttcatag atatcatccg atttgttcaa
      601 ataatttgag ttttgtcgaa taattactct tcgatttgtg atttctatct agatctggtg
      661 ttagtttcta gtttgtgcga tcgaatttgt cgattaatct gagtttttct gattaacagc
      721 accatggcaa aggatgtgga agccgttccc ggagaaggat ttcagacaag agactatcaa
      781 gatccgccac cagctccgtt tattgatgga gcggagctaa agaagtggtc tttctacaga
      841 gcagttatcg cagagttcgt agccactctc ctcttcttat acatcaccgt tttgacagtc
      901 atcggttaca agattcagtc cgatactgat gccggtggcg tagattgcgg cggagttgga
      961 atcctcggta tcgcttgggc ctttggtggt atgatcttca tcctcgtcta ctgcaccgcc
     1021 ggtatctctg gtggtcacat taacccagcg gtgacatttg ggctattctt ggcacgtaaa
     1081 gtgtcgttac ctagggccct attgtacata atcgctcagt gtttgggtgc gatttgtgga
     1141 gttggttttg tcaaagcctt ccaaagctct tactacaccc gttacggagg tggagccaac
     1201 tctctagccg atggctacag cacagggacc ggtctagccg cagagatcat tggtactttc
     1261 gttcttgtct acaccgtatt ctctgccact gaccccaaac gtagtgccag agactcccac
     1321 gttccggtgt tggcgccact tccaatcgga tttgccgtgt tcatggtaca tttggctacc
     1381 attcccatta ccggaaccgg aattaacccg gcaaggagtt tcggagctgc cgtaatctac
     1441 aacaagagca agccatggga tgaccactgg atattttggg ttggaccatt cattggagct
     1501 gcgatagctg cattctacca ccaattcgtt ctaagagctt caggttctaa gtctcttgga
     1561 tcattcagaa gtgctgccaa cgtcggatcc atggtgagca agggcgagga ggataacatg
     1621 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac
     1681 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
     1741 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc
     1801 atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg
     1861 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
     1921 accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc
     1981 ggcaccaact tcccctccga cggccccgta atgcagaata agaccatggg ctgggaggcc
     2041 tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg
     2101 aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag
     2161 cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac
     2221 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc
     2281 atggaggagc tgtacaagta aagcggccgc ccggctgcag atcgttcaaa catttggcaa
     2341 taaagtttct taagattgaa tcctgttgcc ggtcttgcga tgattatcat ataatttctg
     2401 ttgaattacg ttaagcatgt aataattaac atgtaatgca tgacgttatt tatgagatgg
     2461 gtttttatga ttagagtccc gcaattatac atttaatacg cgatagaaaa caaaatatag
     2521 cgcgcaaact aggataaatt atcgcgcgcg gtgtcatcta tgttactaga tcggggcgcg
     2581 ccctcgagga agacgtaatg ggtccccaga tttgcctttt caatttcaga aagaatgcta
     2641 acccacagat ggttagagag gcttacgcag cagctctcat caagacgatc tacccgagca
     2701 ataatctcca ggaaatcaaa taccttccca agaaggttaa agatgcagtc aaaagattca
     2761 ggactaactg catcaagaac acagagaaag atatatttct caagatcaga agtactattc
     2821 cagtatggac gattcaaggc ttgcttcaca aaccaaggca agtaatagag attggagtct
     2881 ctaaaaaggt agttcccact gaatcaaagg ccatggagtc aaagattcaa atagaggacc
     2941 taacagaact cgccgtaaag actggcgaac agttcataca gagtctctta cgactcaatg
     3001 acaagaagaa aatcttcgtc aacatggtgg agcacgacac acttgtctac tccaaaaata
     3061 tcaaagatac agtctcagaa gaccaaaggg caattgagac ttttcaacaa agggtaatat
     3121 ccggaaacct cctcggattc cattgcccag ctatctgtca ctttattgtg aagatagtgg
     3181 aaaaggaagg tggctcctac aaatgccatc attgcgataa aggaaaggcc atcgttgaag
     3241 atgcctctgc cgacagtggt cccaaagatg gacccccacc cacgaggagc atcgtggaaa
     3301 aagaagacgt tccaaccacg tcttcaaagc aagtggattg atgtgatatc tccactgacg
     3361 taagggatga cgcacaatcc cactatcctt cgcaagaccc ttcctctata taaggaagtt
     3421 catttcattt ggagagaaca cgggggactc tagattgaat agactggcct gtctacacca
     3481 tggtgagtaa aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg
     3541 gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg
     3601 gaaaacttac ccttaaattt atttgcacta ctggaaaact acctgttcca tggccaaccc
     3661 tggtcaccac cctgacctac gtaagtttct gcttctacct ttgatatata tataataatt
     3721 atcattaatt agtagtaata taatatttca aatatttttt tcaaaataaa agaatgtagt
     3781 atatagcaat tgcttttctg tagtttataa gtgtgtatat tttaatttat aacttttcta
     3841 atatatgacc aaaatttgtt gatgtgcagg gcgtgcagtg cttctcccgt taccctgatc
     3901 atatgaagcg gcacgacttc ttcaagagcg ccatgcctga gggatacgtg caggagagga
     3961 ccatcttctt caaggacgac gggaactaca agacacgtgc tgaagtcaag tttgagggtg
     4021 acaccctcgt caacaggatc gagcttaagg gaatcgattt caaggaggac ggaaacatcc
     4081 tcggccacaa gttggaatac aactacaact cccacaacgt atacatcatg gccgacaagc
     4141 aaaagaacgg catcaaagcc aacttcaaga cccgccacaa catcgaggac ggcggcgtgc
     4201 aactcgctga tcattatcaa caaaatactc caattggcga tggccctgtc cttttaccag
     4261 acaaccatta cctgtccaca caatctgccc tttcgaaaga tcccaacgaa aagagagatc
     4321 acatggtcct tcttgagttt gtaacagctg ctgggattac acatggcatg gatgaactat
     4381 acaaacctaa gaaaaagcgt aaggtcgagg acccttaatg tcaagccatt ctctcgcaga
     4441 tgatgttcac tttgtgtttt acttccttta tgcattcaca gcaataaaag aaagaaatct
     4501 ccatcgcttt tggttttctt ctctgtctta agttagtcgt tttcgtgtct aatctattac
     4561 ttatcattgt aatagactct tcttctattg agatttgaat ataaactaaa acacattcca
     4621 ttttactgtg ttctcaacat tcagaatgca aacggactaa ccgtagtgac tcggcttgag
     4681 tcttcgggct agagcggact agttctagag cggccgccac cgcggtggag ctccagcttt
     4741 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     4801 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     4861 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     4921 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     4981 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5041 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5101 gaatcagggg ataacgcagg aaagaacatg aaggccttga caggatatat tggcgggtaa
     5161 actaagtcgc tgtatgtgtt tgtttgagat ctcatgtgag caaaaggcca gcaaaaggcc
     5221 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5281 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5341 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5401 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5461 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5521 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5581 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5641 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5701 tttggtatct gtgctctgct gaagccagtt accttcggaa gaagagttgg tagctcttga
     5761 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5821 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5881 tggaacgaaa actcacgtta agggattttg gtcatgaatt cttgttagac attatttgcc
     5941 gactaccttg gtgatctcgc ctttcacgta gtgaacaaat tcttccaact gatctgcgcg
     6001 cgaggccaag cgatcttctt gtccaagata agcctgccta gcttcaagta tgacgggctg
     6061 atactgggcc ggcaggcgct ccattgccca gtcggcagcg acatccttcg gcgcgatttt
     6121 gccggttact gcgctgtacc aaatgcggga caacgtaagc actacatttc gctcatcgcc
     6181 agcccagtcg ggcggcgagt tccatagcgt taaggtttca tttagcgcct caaatagatc
     6241 ctgttcagga accggatcaa agagttcctc cgccgctgga cctaccaagg caacgctatg
     6301 ttctcttgct tttgtcagca agatagccag atcaatgtcg atcgtggctg gctcgaagat
     6361 acctgcaaga atgtcattgc gctgccattc tccaaattgc agttcgcgct tagctggata
     6421 acgccacgga atgatgtcgt cgtgcacaac aatggtgact tctacagcgc ggagaatctc
     6481 gctctctcca ggggaagccg aagtttccaa aaggtcgttg atcaaagctc gccgcgttgt
     6541 ttcatcaagc cttacggtca ccgtaaccag caaatcaata tcactgtgtg gcttcaggcc
     6601 gccatccact gcggagccgt acaaatgtac ggccagcaac gtcggttcga gatggcgctc
     6661 gatgacgcca actacctctg atagttgagt cgatacttcg gcgatcaccg cttccctcat
     6721 gatgtttaac tcctgaatta agccgcgccg cgaagcggtg tcggcttgaa tgaattgtta
     6781 ggcgtcatcc tgtgctcccg agaaccagta ccagtacatc gctgtttcgt tcgagacttg
     6841 aggtctagtt ttatacgtga acaggtcaat gccgccgaga gtaaagccac attttgcgta
     6901 caaattgcag gcaggtacat tgttcgtttg tgccatgatg atatattttt atcttgtgca
     6961 atgtaacatc agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct
     7021 gagttgaagg atcagatcac gcatcttccc gacaacgcag accgttccgt ggcaaagcaa
     7081 aagttcaaaa tcaccaactg gtccacctac aacaaagctc tcatcaaccg tggctccctc
     7141 actttctggc tggatgatgg ggcgattcag gagatcccca tccaacagcc cgccgtcgag
     7201 cgggcttttt tatccccgga agcctgtgga tagagggtag ttatccacgt gaaaccgcta
     7261 atgccccgca aagccttgat tcacggggct ttccggcccg ctccaaaaac tatccacgtg
     7321 aaatcgctaa tcagggtacg tgaaatcgct aatcggagta cgtgaaatcg ctaataaggt
     7381 cacgtgaaat cgctaatcaa aaaggcacgt gagaacgcta atagcccttt cagatcaaca
     7441 gcttgcaaac acccctcgct ccggcaagta gttacagcaa gtagtatgtt caattagctt
     7501 ttcaattatg aatatatata tcaattattg gtcgcccttg gcttgtggac aatgcgctac
     7561 gcgcaccggc tccgcccgtg gacaaccgca agcggttgcc caccgtcgag cgcctttgcc
     7621 cacaacccgg cggccggccg caacagatcg ttttataaat tttttttttt gaaaaagaaa
     7681 aagcccgaaa ggcgg
//
