LOCUS       sequence-387023-        5822 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proG1090::AmtR-VP16-19St).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5822)
  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 5822)
  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: JAB2156
FEATURES             Location/Qualifiers
     source          1..5822
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      169..604
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     CDS             1062..1850
                     /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      1941..2529
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2430..2449
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2620..2644
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     2749..2766
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2883..2904
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2919..2924,2925..2942,2943..2949)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2957..2973
                     /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)."
     primer_bind     2962..2984
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2981..2997
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2981..2997
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3016..3036
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3018..3036
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    3118..3294
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(3350..3370)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(3371..3604)
                     /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"
     promoter        complement(4289..4335)
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     primer_bind     complement(4324..4344)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        4661..4840
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             4882..5433
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     terminator      5453..5705
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    5775..5799
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 ccggcagctc ggcacaaaat caccactcga tacaggcagc cagagtctaa ttccggggat
       61 cggaaatcca gaagcccgag aggttgccgc ctttcgggct ttttcttttt caaaaaaaaa
      121 aatttataaa acgatctgtt gcggccggcc gccgggttgt gggcaaaggc gctcgacggt
      181 gggcaaccgc ttgcggttgt ccacgggcgg agccggtgcg cgtagcgcat tgtccacaag
      241 ccaagggcga ccaataattg atatatatat tcataattga aaagctaatt gaacatacta
      301 cttgctgtaa ctacttgccg gagcgagggg tgtttgcaag ctgttgatct gaaagggcta
      361 ttagcgttct cacgtgcctt tttgattagc gatttcacgt gaccttatta gcgatttcac
      421 gtactccgat tagcgatttc acgtaccctg attagcgatt tcacgtggat agtttttgga
      481 gcgggccgga aagccccgtg aatcaaggct ttgcggggca ttagcggttt cacgtggata
      541 actaccctct atccacaggc ttccggggat aaaaaagccc gctcgacggc gggctgttgg
      601 atggggatct cctgaatcgc cccatcatcc agccagaaag tgagggagcc acggttgatg
      661 agagctttgt tgtaggtgga ccagttggtg attttgaact tttgctttgc cacggaacgg
      721 tctgcgttgt cgggaagatg cgtgatctga tccttcaact cagcaaaagt tcgatttatt
      781 caacaaagcc acgttgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata
      841 tcatcatggc acaaacgaac aatgtacctg cctgcaattt gtacgcaaaa tgtggcttta
      901 ctctcggcgg cattgacctg ttcacgtata aaactagacc tcaagtctcg aacgaaacag
      961 cgatgtactg gtactggttc tcgggagcac aggatgacgc ctaacaattc attcaagccg
     1021 acaccgcttc gcggcgcggc ttaattcagg agttaaacat catgagggaa gcggtgatcg
     1081 ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga
     1141 cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag ccacacagtg
     1201 atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga
     1261 tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc cgcgctgtag
     1321 aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct aagcgcgaac
     1381 tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag ccagccacga
     1441 tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt gccttggtag
     1501 gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt gaggcgctaa
     1561 atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag cgaaatgtag
     1621 tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg
     1681 tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc atacttgaag
     1741 ctaggcaggc ttatcttgga caagaagatc gcttggcctc gcgcgcagat cagttggaag
     1801 aatttgttca ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa tgtctaacaa
     1861 gaattcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     1921 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     1981 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     2041 cttccgaagg taactggctt cagcagagca cagataccaa atactgttct tctagtgtag
     2101 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     2161 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     2221 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     2281 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     2341 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     2401 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     2461 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     2521 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     2581 gctcacatga gatctcaaac aaacacatac agcgacttag tttacccgcc aatatatcct
     2641 gtcaaggcct tcatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     2701 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     2761 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     2821 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     2881 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     2941 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     3001 gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctgga gctccaccgc
     3061 ggtggcggcc gctctagaac tagtccgctc tagcccgaag acgtagtatc aagcatcgat
     3121 ctggatttta gtactggatt ttggttttag gaattagaaa ttttattgat agaagtattt
     3181 tacaaataca aatacatact aagggtttct tatatgctca acacatgagc gaaaccctat
     3241 aagaacccta atttccctta tcgggaaact actcacacat tatttatgga gaaaatagag
     3301 agagatagat ttgtagagag agactggtga tttcagcgta ccgacactaa accttcctct
     3361 tcttcttcgg cccaccgtac tcgtcaattc caagggcatc ggtaaacatc tgctcaaact
     3421 cgaagtcggc cgtatccaga gcgccgtagg gggcggagtc gtggggggta aatcccggcc
     3481 ccggggaatc cccgtccccc aacatgtcca gatcgaaatc gtctagcgcg tcggcatgcg
     3541 ccatcgccac gtcctcgccg tctaagtgga gctcgtcccc caggctgacg tcggtcgggg
     3601 gggccttggc gtcggcctgc ttgatcagtt ccagtgtctt ttccactctg tcggcaggca
     3661 gaggtgctcc cagcacagcc agggaggcgt cagccagcat aatggctgtc tcgggcagag
     3721 agtcggcgct cagagggcta gggatcttgc cgtcgttccg ccgcatctcg atcacgctca
     3781 tggtgatgtg gaagggcagt tcggctctgg ggtcgtcgcc cacaatctcg gtggccagat
     3841 ctctgaacac gttggtcagg gcctcgcgct ggctgtggta ctcggcgaat tcctcagagc
     3901 ccacgatggg cagctggtac agccggccca cgttccactt ggtgctcagc agcagccgca
     3961 cttcgctggc cacgatagcc cacagtctca tttcgggtcc ggcgtccagg gtagacagat
     4021 cctcggccag cacggtggag ggttccacgg tgcttttcag cagggtcagg aagatctcgg
     4081 tcttgctggg gaagtggtag tacaggctgg cctgtctgat gcccacagcg tcggcaatct
     4141 ggtgggtgct ggtggtggca aagccctgtc tggtgaacag ctcggcgctg gcatccagaa
     4201 tctcttcccg ggggttcttg ccggctctcc gaggagcaga tcttctgggt ctgcccacgg
     4261 cgccagccat ggtggggatc ccagcgtgtc ctctccaaat gaaatgaact tccttatata
     4321 gaggaagggt cttgcgaagg atagtgggat tgtgcgtcat cccttacgtc agtggagatg
     4381 gatcctctag aggcacgtgg cggcacgtgg cggcacgtgg cggcacgtgg caagcttgga
     4441 tccacgatat cctgcaggca tgcaagctta gcttgagctt ggatcagatt gtcgtttccc
     4501 gccttcagtt taaactacat tagaagcacg tcttcacacg ttaaactttg agctcgtccg
     4561 atagtcgaca tgttatcgat catgagcgga gaattaaggg agtcacgtta tgacccccgc
     4621 cgatgacgcg ggacaagccg ttttacgttt ggaactgaca gaaccgcaac gttgaaggag
     4681 ccactgagcc gcgggtttct ggagtttaat gagctaagca catacgtcag aaaccattat
     4741 tgcgcgttca aaagtcgcct aaggtcacta tcagctagca aatatttctt gtcaaaaatg
     4801 ctccactgac gttccataaa ttcccctcgg tatccaatta gagtctcata ttcactctca
     4861 actcgatcga ggggatctac catgagccca gaacgacgcc cggccgacat ccgccgtgcc
     4921 accgaggcgg acatgccggc ggtctgcacc atcgtcaacc actacatcga gacaagcacg
     4981 gtcaacttcc gtaccgagcc gcaggaaccg caggagtgga cggacgacct cgtccgtctg
     5041 cgggagcgct atccctggct cgtcgccgag gtggacggcg aggtcgccgg catcgcctac
     5101 gcgggtccct ggaaggcacg caacgcctac gactggacgg ccgagtcgac cgtgtacgtg
     5161 tccccccgcc accagcggac gggactgggc tccacgctct acacccacct gctgaagtcc
     5221 ctggaggcac agggcttcaa gagcgtggtc gctgtcatcg ggctgcccaa cgacccgagc
     5281 gtgcgcatgc acgaggcgct cggatatgcc ccccgcggca tgctgcgggc ggccggcttc
     5341 aagcacggga actggcatga cgtgggtttc tggcagctgg acttcagcct gccggtgccg
     5401 ccccgtccgg tcctgcccgt caccgaaatc tgatgacccc tagagtcaag cagatcgttc
     5461 aaacatttgg caataaagtt tcttaagatt gaatcctgtt gccggtcttg cgatgattat
     5521 catataattt ctgttgaatt acgttaagca tgtaataatt aacatgtaat gcatgacgtt
     5581 atttatgaga tgggttttta tgattagagt cccgcaatta tacatttaat acgcgataga
     5641 aaacaaaata tagcgcgcaa actaggataa attatcgcgc gcggtgtcat ctatgttact
     5701 agatcgaccg gcatgcaagc tgattacatt aaaaacgtcc gcaatgtgtt attaagttgt
     5761 ctaagcgtca atttgtttac accacaatat atcctgccac cagccagcca acagctcccc
     5821 ga
//
