LOCUS       sequence-387095-        5651 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proG1090::RpaR-NLS_19St).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5651)
  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 5651)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: JAB2518
FEATURES             Location/Qualifiers
     source          1..5651
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    5..29
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(99..351)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(371..922)
                     /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(964..1143)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     1460..1480
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        1469..1515
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             2263..2283
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2339..2515
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(2597..2617)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2597..2615)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2636..2652)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2636..2652)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2649..2671)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2660..2676
                     /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(2684..2690,2691..2708,2709..2714))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2729..2750
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2867..2884)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2989..3013
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(3104..3692)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3184..3203)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3783..4571)
                     /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      5029..5464
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 ctggtggcag gatatattgt ggtgtaaaca aattgacgct tagacaactt aataacacat
       61 tgcggacgtt tttaatgtaa tcagcttgca tgccggtcga tctagtaaca tagatgacac
      121 cgcgcgcgat aatttatcct agtttgcgcg ctatattttg ttttctatcg cgtattaaat
      181 gtataattgc gggactctaa tcataaaaac ccatctcata aataacgtca tgcattacat
      241 gttaattatt acatgcttaa cgtaattcaa cagaaattat atgataatca tcgcaagacc
      301 ggcaacagga ttcaatctta agaaacttta ttgccaaatg tttgaacgat ctgcttgact
      361 ctaggggtca tcagatttcg gtgacgggca ggaccggacg gggcggcacc ggcaggctga
      421 agtccagctg ccagaaaccc acgtcatgcc agttcccgtg cttgaagccg gccgcccgca
      481 gcatgccgcg gggggcatat ccgagcgcct cgtgcatgcg cacgctcggg tcgttgggca
      541 gcccgatgac agcgaccacg ctcttgaagc cctgtgcctc cagggacttc agcaggtggg
      601 tgtagagcgt ggagcccagt cccgtccgct ggtggcgggg ggacacgtac acggtcgact
      661 cggccgtcca gtcgtaggcg ttgcgtgcct tccagggacc cgcgtaggcg atgccggcga
      721 cctcgccgtc cacctcggcg acgagccagg gatagcgctc ccgcagacgg acgaggtcgt
      781 ccgtccactc ctgcggttcc tgcggctcgg tacggaagtt gaccgtgctt gtctcgatgt
      841 agtggttgac gatggtgcag accgccggca tgtccgcctc ggtggcacgg cggatgtcgg
      901 ccgggcgtcg ttctgggctc atggtagatc ccctcgatcg agttgagagt gaatatgaga
      961 ctctaattgg ataccgaggg gaatttatgg aacgtcagtg gagcattttt gacaagaaat
     1021 atttgctagc tgatagtgac cttaggcgac ttttgaacgc gcaataatgg tttctgacgt
     1081 atgtgcttag ctcattaaac tccagaaacc cgcggctcag tggctccttc aacgttgcgg
     1141 ttctgtcagt tccaaacgta aaacggcttg tcccgcgtca tcggcggggg tcataacgtg
     1201 actcccttaa ttctccgctc atgatcgata acatgtcgac tatcggacga gctcaaagtt
     1261 taacgtgtga agacgtgctt ctaatgtagt ttaaactgaa ggcgggaaac gacaatctga
     1321 tccaagctca agctaagctt gcatgcctgc aggatatcgt ggatccaagc ttgccacgtg
     1381 ccgccacgtg ccgccacgtg ccgccacgtg cctctagagg atccatctcc actgacgtaa
     1441 gggatgacgc acaatcccac tatccttcgc aagacccttc ctctatataa ggaagttcat
     1501 ttcatttgga gaggacacgc tgggatcccc accatgattg tgggtgaaga tcagctgtgg
     1561 ggtcgtcgta cactggaatt tgttgatagc gttgaacgtc tggaagcacc ggcactgatt
     1621 agccgttttg aaagcctgat tgcaagctgt ggttttaccg cctatatcat ggcaggtctg
     1681 ccgagccgta atgccggtct gccggaactg accctggcaa atggttggcc tcgtgattgg
     1741 tttgatctgt atgttagcga aaactttagc gcagttgatc cggttccgcg ttatggtgca
     1801 accactgttc atccgtttgt ttggagtgat gcaccgtatg atcgtgaccg tgatcaggca
     1861 gcacatcgtg ttatgacccg tgcagcagaa tttggtctgg ttgaaggtta ttgtattccg
     1921 ctgcattacg atgatggtag cgcagcaatt agtatggcag gtgaagatcc tgatctgagt
     1981 ccggcagccc gtggtgtaat gcagctggtt agcatttatg cacatagccg tctgcgtgta
     2041 ctgagccgtc cgaaaccgat tcgtcgtaat cgtctgacac cgcgtgaatg tgaaattctg
     2101 cagtgggcag cacagggtaa aaccgcatgg gaaattagcg ttattctgtg tattaccgaa
     2161 cgcaccgtta aatttcatct gattgaagca gcacgtaaac tggatgcagc aaatcgtacc
     2221 gcagcagttg caaaagcact gacactgggt ctgattcgtc tgccgaagaa gaagaggaag
     2281 gtttagtgtc ggtacgctga aatcaccagt ctctctctac aaatctatct ctctctattt
     2341 tctccataaa taatgtgtga gtagtttccc gataagggaa attagggttc ttatagggtt
     2401 tcgctcatgt gttgagcata taagaaaccc ttagtatgta tttgtatttg taaaatactt
     2461 ctatcaataa aatttctaat tcctaaaacc aaaatccagt actaaaatcc agatcgatgc
     2521 ttgatactac gtcttcgggc tagagcggac tagttctaga gcggccgcca ccgcggtgga
     2581 gctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat
     2641 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     2701 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     2761 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     2821 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     2881 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     2941 ggttatccac agaatcaggg gataacgcag gaaagaacat gaaggccttg acaggatata
     3001 ttggcgggta aactaagtcg ctgtatgtgt ttgtttgaga tctcatgtga gcaaaaggcc
     3061 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3121 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3181 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3241 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3301 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3361 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3421 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3481 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3541 gaagaacagt atttggtatc tgtgctctgc tgaagccagt taccttcgga agaagagttg
     3601 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3661 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     3721 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaat tcttgttaga
     3781 cattatttgc cgactacctt ggtgatctcg cctttcacgt agtgaacaaa ttcttccaac
     3841 tgatctgcgc gcgaggccaa gcgatcttct tgtccaagat aagcctgcct agcttcaagt
     3901 atgacgggct gatactgggc cggcaggcgc tccattgccc agtcggcagc gacatccttc
     3961 ggcgcgattt tgccggttac tgcgctgtac caaatgcggg acaacgtaag cactacattt
     4021 cgctcatcgc cagcccagtc gggcggcgag ttccatagcg ttaaggtttc atttagcgcc
     4081 tcaaatagat cctgttcagg aaccggatca aagagttcct ccgccgctgg acctaccaag
     4141 gcaacgctat gttctcttgc ttttgtcagc aagatagcca gatcaatgtc gatcgtggct
     4201 ggctcgaaga tacctgcaag aatgtcattg cgctgccatt ctccaaattg cagttcgcgc
     4261 ttagctggat aacgccacgg aatgatgtcg tcgtgcacaa caatggtgac ttctacagcg
     4321 cggagaatct cgctctctcc aggggaagcc gaagtttcca aaaggtcgtt gatcaaagct
     4381 cgccgcgttg tttcatcaag ccttacggtc accgtaacca gcaaatcaat atcactgtgt
     4441 ggcttcaggc cgccatccac tgcggagccg tacaaatgta cggccagcaa cgtcggttcg
     4501 agatggcgct cgatgacgcc aactacctct gatagttgag tcgatacttc ggcgatcacc
     4561 gcttccctca tgatgtttaa ctcctgaatt aagccgcgcc gcgaagcggt gtcggcttga
     4621 atgaattgtt aggcgtcatc ctgtgctccc gagaaccagt accagtacat cgctgtttcg
     4681 ttcgagactt gaggtctagt tttatacgtg aacaggtcaa tgccgccgag agtaaagcca
     4741 cattttgcgt acaaattgca ggcaggtaca ttgttcgttt gtgccatgat gatatatttt
     4801 tatcttgtgc aatgtaacat cagagatttt gagacacaac gtggctttgt tgaataaatc
     4861 gaacttttgc tgagttgaag gatcagatca cgcatcttcc cgacaacgca gaccgttccg
     4921 tggcaaagca aaagttcaaa atcaccaact ggtccaccta caacaaagct ctcatcaacc
     4981 gtggctccct cactttctgg ctggatgatg gggcgattca ggagatcccc atccaacagc
     5041 ccgccgtcga gcgggctttt ttatccccgg aagcctgtgg atagagggta gttatccacg
     5101 tgaaaccgct aatgccccgc aaagccttga ttcacggggc tttccggccc gctccaaaaa
     5161 ctatccacgt gaaatcgcta atcagggtac gtgaaatcgc taatcggagt acgtgaaatc
     5221 gctaataagg tcacgtgaaa tcgctaatca aaaaggcacg tgagaacgct aatagccctt
     5281 tcagatcaac agcttgcaaa cacccctcgc tccggcaagt agttacagca agtagtatgt
     5341 tcaattagct tttcaattat gaatatatat atcaattatt ggtcgccctt ggcttgtgga
     5401 caatgcgcta cgcgcaccgg ctccgcccgt ggacaaccgc aagcggttgc ccaccgtcga
     5461 gcgcctttgc ccacaacccg gcggccggcc gcaacagatc gttttataaa tttttttttt
     5521 tgaaaaagaa aaagcccgaa aggcggcaac ctctcgggct tctggatttc cgatccccgg
     5581 aattagactc tggctgcctg tatcgagtgg tgattttgtg ccgagctgcc ggtcggggag
     5641 ctgttggctg g
//
