LOCUS       sequence-389758-        6002 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proG1090::LacI-NLS-19St).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6002)
  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 6002)
  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: JAB2657
FEATURES             Location/Qualifiers
     source          1..6002
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    10..34
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(104..356)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(376..927)
                     /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(969..1148)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     1465..1485
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        1474..1520
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             1539..2618
                     /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(1558..1577)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2619..2639
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2695..2871
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(2953..2973)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2953..2971)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2992..3008)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2992..3008)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3005..3027)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3016..3032
                     /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(3040..3046,3047..3064,3065..3070))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3085..3106
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3223..3240)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3345..3369
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(3460..4048)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3540..3559)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4139..4927)
                     /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      5385..5820
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 gctggctggt ggcaggatat attgtggtgt aaacaaattg acgcttagac aacttaataa
       61 cacattgcgg acgtttttaa tgtaatcagc ttgcatgccg gtcgatctag taacatagat
      121 gacaccgcgc gcgataattt atcctagttt gcgcgctata ttttgttttc tatcgcgtat
      181 taaatgtata attgcgggac tctaatcata aaaacccatc tcataaataa cgtcatgcat
      241 tacatgttaa ttattacatg cttaacgtaa ttcaacagaa attatatgat aatcatcgca
      301 agaccggcaa caggattcaa tcttaagaaa ctttattgcc aaatgtttga acgatctgct
      361 tgactctagg ggtcatcaga tttcggtgac gggcaggacc ggacggggcg gcaccggcag
      421 gctgaagtcc agctgccaga aacccacgtc atgccagttc ccgtgcttga agccggccgc
      481 ccgcagcatg ccgcgggggg catatccgag cgcctcgtgc atgcgcacgc tcgggtcgtt
      541 gggcagcccg atgacagcga ccacgctctt gaagccctgt gcctccaggg acttcagcag
      601 gtgggtgtag agcgtggagc ccagtcccgt ccgctggtgg cggggggaca cgtacacggt
      661 cgactcggcc gtccagtcgt aggcgttgcg tgccttccag ggacccgcgt aggcgatgcc
      721 ggcgacctcg ccgtccacct cggcgacgag ccagggatag cgctcccgca gacggacgag
      781 gtcgtccgtc cactcctgcg gttcctgcgg ctcggtacgg aagttgaccg tgcttgtctc
      841 gatgtagtgg ttgacgatgg tgcagaccgc cggcatgtcc gcctcggtgg cacggcggat
      901 gtcggccggg cgtcgttctg ggctcatggt agatcccctc gatcgagttg agagtgaata
      961 tgagactcta attggatacc gaggggaatt tatggaacgt cagtggagca tttttgacaa
     1021 gaaatatttg ctagctgata gtgaccttag gcgacttttg aacgcgcaat aatggtttct
     1081 gacgtatgtg cttagctcat taaactccag aaacccgcgg ctcagtggct ccttcaacgt
     1141 tgcggttctg tcagttccaa acgtaaaacg gcttgtcccg cgtcatcggc gggggtcata
     1201 acgtgactcc cttaattctc cgctcatgat cgataacatg tcgactatcg gacgagctca
     1261 aagtttaacg tgtgaagacg tgcttctaat gtagtttaaa ctgaaggcgg gaaacgacaa
     1321 tctgatccaa gctcaagcta agcttgcatg cctgcaggat atcgtggatc caagcttgcc
     1381 acgtgccgcc acgtgccgcc acgtgccgcc acgtgcctct agaggatcca tctccactga
     1441 cgtaagggat gacgcacaat cccactatcc ttcgcaagac ccttcctcta tataaggaag
     1501 ttcatttcat ttggagagga cacgctggga tccccaccat gaaaccagta acgttatacg
     1561 atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca
     1621 gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca
     1681 ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca
     1741 cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg
     1801 atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta
     1861 aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc
     1921 tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc
     1981 ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgag gacggtacgc
     2041 gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc
     2101 cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca
     2161 atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac
     2221 aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc
     2281 agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata
     2341 tctcggtagt gggatacgac gataccgaag atagctcatg ttatatcccg ccgttaacca
     2401 ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct
     2461 ctcagggcca ggcggtgaag ggcaatcagc tgttgccagt ctcactggtg aaaagaaaaa
     2521 ccaccctggc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     2581 agctggcacg acaggtttcc cgactggaaa gcgggcagcc gaagaagaag aggaaggttt
     2641 agtgtcggta cgctgaaatc accagtctct ctctacaaat ctatctctct ctattttctc
     2701 cataaataat gtgtgagtag tttcccgata agggaaatta gggttcttat agggtttcgc
     2761 tcatgtgttg agcatataag aaacccttag tatgtatttg tatttgtaaa atacttctat
     2821 caataaaatt tctaattcct aaaaccaaaa tccagtacta aaatccagat cgatgcttga
     2881 tactacgtct tcgggctaga gcggactagt tctagagcgg ccgccaccgc ggtggagctc
     2941 cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct
     3001 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     3061 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     3121 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3181 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3241 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3301 atccacagaa tcaggggata acgcaggaaa gaacatgaag gccttgacag gatatattgg
     3361 cgggtaaact aagtcgctgt atgtgtttgt ttgagatctc atgtgagcaa aaggccagca
     3421 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3481 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3541 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3601 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3661 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3721 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3781 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3841 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3901 aacagtattt ggtatctgtg ctctgctgaa gccagttacc ttcggaagaa gagttggtag
     3961 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4021 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4081 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgaattctt gttagacatt
     4141 atttgccgac taccttggtg atctcgcctt tcacgtagtg aacaaattct tccaactgat
     4201 ctgcgcgcga ggccaagcga tcttcttgtc caagataagc ctgcctagct tcaagtatga
     4261 cgggctgata ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg
     4321 cgattttgcc ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct
     4381 catcgccagc ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa
     4441 atagatcctg ttcaggaacc ggatcaaaga gttcctccgc cgctggacct accaaggcaa
     4501 cgctatgttc tcttgctttt gtcagcaaga tagccagatc aatgtcgatc gtggctggct
     4561 cgaagatacc tgcaagaatg tcattgcgct gccattctcc aaattgcagt tcgcgcttag
     4621 ctggataacg ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga
     4681 gaatctcgct ctctccaggg gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc
     4741 gcgttgtttc atcaagcctt acggtcaccg taaccagcaa atcaatatca ctgtgtggct
     4801 tcaggccgcc atccactgcg gagccgtaca aatgtacggc cagcaacgtc ggttcgagat
     4861 ggcgctcgat gacgccaact acctctgata gttgagtcga tacttcggcg atcaccgctt
     4921 ccctcatgat gtttaactcc tgaattaagc cgcgccgcga agcggtgtcg gcttgaatga
     4981 attgttaggc gtcatcctgt gctcccgaga accagtacca gtacatcgct gtttcgttcg
     5041 agacttgagg tctagtttta tacgtgaaca ggtcaatgcc gccgagagta aagccacatt
     5101 ttgcgtacaa attgcaggca ggtacattgt tcgtttgtgc catgatgata tatttttatc
     5161 ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttgttgaa taaatcgaac
     5221 ttttgctgag ttgaaggatc agatcacgca tcttcccgac aacgcagacc gttccgtggc
     5281 aaagcaaaag ttcaaaatca ccaactggtc cacctacaac aaagctctca tcaaccgtgg
     5341 ctccctcact ttctggctgg atgatggggc gattcaggag atccccatcc aacagcccgc
     5401 cgtcgagcgg gcttttttat ccccggaagc ctgtggatag agggtagtta tccacgtgaa
     5461 accgctaatg ccccgcaaag ccttgattca cggggctttc cggcccgctc caaaaactat
     5521 ccacgtgaaa tcgctaatca gggtacgtga aatcgctaat cggagtacgt gaaatcgcta
     5581 ataaggtcac gtgaaatcgc taatcaaaaa ggcacgtgag aacgctaata gccctttcag
     5641 atcaacagct tgcaaacacc cctcgctccg gcaagtagtt acagcaagta gtatgttcaa
     5701 ttagcttttc aattatgaat atatatatca attattggtc gcccttggct tgtggacaat
     5761 gcgctacgcg caccggctcc gcccgtggac aaccgcaagc ggttgcccac cgtcgagcgc
     5821 ctttgcccac aacccggcgg ccggccgcaa cagatcgttt tataaatttt tttttttgaa
     5881 aaagaaaaag cccgaaaggc ggcaacctct cgggcttctg gatttccgat ccccggaatt
     5941 agactctggc tgcctgtatc gagtggtgat tttgtgccga gctgccggtc ggggagctgt
     6001 tg
//
