LOCUS       sequence-387107-        6245 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proG1090::LacI-VP16-19St).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6245)
  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 6245)
  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: JAB2618
FEATURES             Location/Qualifiers
     source          1..6245
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    14..38
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(108..360)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(380..931)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     promoter        complement(973..1152)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     1469..1489
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        1478..1524
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             1543..2622
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(1562..1581)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2632..2865
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDTADFEFEQMFTDALGIDEYGG"
     CDS             2866..2886
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2942..3118
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(3200..3220)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3200..3218)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3239..3255)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3239..3255)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3252..3274)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3263..3279
                     /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(3287..3293,3294..3311,3312..3317))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3332..3353
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3470..3487)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3592..3616
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(3707..4295)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3787..3806)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4386..5174)
                     /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      5632..6067
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 gttggctggc tggtggcagg atatattgtg gtgtaaacaa attgacgctt agacaactta
       61 ataacacatt gcggacgttt ttaatgtaat cagcttgcat gccggtcgat ctagtaacat
      121 agatgacacc gcgcgcgata atttatccta gtttgcgcgc tatattttgt tttctatcgc
      181 gtattaaatg tataattgcg ggactctaat cataaaaacc catctcataa ataacgtcat
      241 gcattacatg ttaattatta catgcttaac gtaattcaac agaaattata tgataatcat
      301 cgcaagaccg gcaacaggat tcaatcttaa gaaactttat tgccaaatgt ttgaacgatc
      361 tgcttgactc taggggtcat cagatttcgg tgacgggcag gaccggacgg ggcggcaccg
      421 gcaggctgaa gtccagctgc cagaaaccca cgtcatgcca gttcccgtgc ttgaagccgg
      481 ccgcccgcag catgccgcgg ggggcatatc cgagcgcctc gtgcatgcgc acgctcgggt
      541 cgttgggcag cccgatgaca gcgaccacgc tcttgaagcc ctgtgcctcc agggacttca
      601 gcaggtgggt gtagagcgtg gagcccagtc ccgtccgctg gtggcggggg gacacgtaca
      661 cggtcgactc ggccgtccag tcgtaggcgt tgcgtgcctt ccagggaccc gcgtaggcga
      721 tgccggcgac ctcgccgtcc acctcggcga cgagccaggg atagcgctcc cgcagacgga
      781 cgaggtcgtc cgtccactcc tgcggttcct gcggctcggt acggaagttg accgtgcttg
      841 tctcgatgta gtggttgacg atggtgcaga ccgccggcat gtccgcctcg gtggcacggc
      901 ggatgtcggc cgggcgtcgt tctgggctca tggtagatcc cctcgatcga gttgagagtg
      961 aatatgagac tctaattgga taccgagggg aatttatgga acgtcagtgg agcatttttg
     1021 acaagaaata tttgctagct gatagtgacc ttaggcgact tttgaacgcg caataatggt
     1081 ttctgacgta tgtgcttagc tcattaaact ccagaaaccc gcggctcagt ggctccttca
     1141 acgttgcggt tctgtcagtt ccaaacgtaa aacggcttgt cccgcgtcat cggcgggggt
     1201 cataacgtga ctcccttaat tctccgctca tgatcgataa catgtcgact atcggacgag
     1261 ctcaaagttt aacgtgtgaa gacgtgcttc taatgtagtt taaactgaag gcgggaaacg
     1321 acaatctgat ccaagctcaa gctaagcttg catgcctgca ggatatcgtg gatccaagct
     1381 tgccacgtgc cgccacgtgc cgccacgtgc cgccacgtgc ctctagagga tccatctcca
     1441 ctgacgtaag ggatgacgca caatcccact atccttcgca agacccttcc tctatataag
     1501 gaagttcatt tcatttggag aggacacgct gggatcccca ccatgaaacc agtaacgtta
     1561 tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag
     1621 gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat
     1681 tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt
     1741 gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc
     1801 gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc
     1861 tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat
     1921 ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta
     1981 tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca tgaggacggt
     2041 acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg
     2101 ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact
     2161 cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt
     2221 caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac
     2281 gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg
     2341 gatatctcgg tagtgggata cgacgatacc gaagatagct catgttatat cccgccgtta
     2401 accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa
     2461 ctctctcagg gccaggcggt gaagggcaat cagctgttgc cagtctcact ggtgaaaaga
     2521 aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     2581 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agggaggagg tgcccccccg
     2641 accgacgtca gcctggggga cgagctccac ttagacggcg aggacgtggc gatggcgcat
     2701 gccgacgcgc tagacgattt cgatctggac atgttggggg acggggattc cccggggccg
     2761 ggatttaccc cccacgactc cgccccctac ggcgctctgg atacggccga cttcgagttt
     2821 gagcagatgt ttaccgatgc ccttggaatt gacgagtacg gtgggccgaa gaagaagagg
     2881 aaggtttagt gtcggtacgc tgaaatcacc agtctctctc tacaaatcta tctctctcta
     2941 ttttctccat aaataatgtg tgagtagttt cccgataagg gaaattaggg ttcttatagg
     3001 gtttcgctca tgtgttgagc atataagaaa cccttagtat gtatttgtat ttgtaaaata
     3061 cttctatcaa taaaatttct aattcctaaa accaaaatcc agtactaaaa tccagatcga
     3121 tgcttgatac tacgtcttcg ggctagagcg gactagttct agagcggccg ccaccgcggt
     3181 ggagctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt
     3241 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     3301 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     3361 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     3421 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     3481 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3541 tacggttatc cacagaatca ggggataacg caggaaagaa catgaaggcc ttgacaggat
     3601 atattggcgg gtaaactaag tcgctgtatg tgtttgtttg agatctcatg tgagcaaaag
     3661 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3721 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3781 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3841 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3901 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3961 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4021 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4081 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4141 ctagaagaac agtatttggt atctgtgctc tgctgaagcc agttaccttc ggaagaagag
     4201 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4261 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4321 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg aattcttgtt
     4381 agacattatt tgccgactac cttggtgatc tcgcctttca cgtagtgaac aaattcttcc
     4441 aactgatctg cgcgcgaggc caagcgatct tcttgtccaa gataagcctg cctagcttca
     4501 agtatgacgg gctgatactg ggccggcagg cgctccattg cccagtcggc agcgacatcc
     4561 ttcggcgcga ttttgccggt tactgcgctg taccaaatgc gggacaacgt aagcactaca
     4621 tttcgctcat cgccagccca gtcgggcggc gagttccata gcgttaaggt ttcatttagc
     4681 gcctcaaata gatcctgttc aggaaccgga tcaaagagtt cctccgccgc tggacctacc
     4741 aaggcaacgc tatgttctct tgcttttgtc agcaagatag ccagatcaat gtcgatcgtg
     4801 gctggctcga agatacctgc aagaatgtca ttgcgctgcc attctccaaa ttgcagttcg
     4861 cgcttagctg gataacgcca cggaatgatg tcgtcgtgca caacaatggt gacttctaca
     4921 gcgcggagaa tctcgctctc tccaggggaa gccgaagttt ccaaaaggtc gttgatcaaa
     4981 gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa ccagcaaatc aatatcactg
     5041 tgtggcttca ggccgccatc cactgcggag ccgtacaaat gtacggccag caacgtcggt
     5101 tcgagatggc gctcgatgac gccaactacc tctgatagtt gagtcgatac ttcggcgatc
     5161 accgcttccc tcatgatgtt taactcctga attaagccgc gccgcgaagc ggtgtcggct
     5221 tgaatgaatt gttaggcgtc atcctgtgct cccgagaacc agtaccagta catcgctgtt
     5281 tcgttcgaga cttgaggtct agttttatac gtgaacaggt caatgccgcc gagagtaaag
     5341 ccacattttg cgtacaaatt gcaggcaggt acattgttcg tttgtgccat gatgatatat
     5401 ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtggctt tgttgaataa
     5461 atcgaacttt tgctgagttg aaggatcaga tcacgcatct tcccgacaac gcagaccgtt
     5521 ccgtggcaaa gcaaaagttc aaaatcacca actggtccac ctacaacaaa gctctcatca
     5581 accgtggctc cctcactttc tggctggatg atggggcgat tcaggagatc cccatccaac
     5641 agcccgccgt cgagcgggct tttttatccc cggaagcctg tggatagagg gtagttatcc
     5701 acgtgaaacc gctaatgccc cgcaaagcct tgattcacgg ggctttccgg cccgctccaa
     5761 aaactatcca cgtgaaatcg ctaatcaggg tacgtgaaat cgctaatcgg agtacgtgaa
     5821 atcgctaata aggtcacgtg aaatcgctaa tcaaaaaggc acgtgagaac gctaatagcc
     5881 ctttcagatc aacagcttgc aaacacccct cgctccggca agtagttaca gcaagtagta
     5941 tgttcaatta gcttttcaat tatgaatata tatatcaatt attggtcgcc cttggcttgt
     6001 ggacaatgcg ctacgcgcac cggctccgcc cgtggacaac cgcaagcggt tgcccaccgt
     6061 cgagcgcctt tgcccacaac ccggcggccg gccgcaacag atcgttttat aaattttttt
     6121 ttttgaaaaa gaaaaagccc gaaaggcggc aacctctcgg gcttctggat ttccgatccc
     6181 cggaattaga ctctggctgc ctgtatcgag tggtgatttt gtgccgagct gccggtcggg
     6241 gagct
//
