LOCUS       sequence-387110-        6494 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids encode parts for syntetic genetic circuit construction in 
            eudicot plants (proG1090::AmtR-TPL(RD)-19St).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6494)
  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 6494)
  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: JAB2858
FEATURES             Location/Qualifiers
     source          1..6494
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    4..28
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(98..350)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(370..921)
                     /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(963..1142)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     1459..1479
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        1468..1514
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             3105..3125
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3181..3357
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(3439..3459)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3439..3457)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3478..3494)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3478..3494)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3491..3513)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3502..3518
                     /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(3526..3532,3533..3550,3551..3556))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3571..3592
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3709..3726)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3831..3855
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(3946..4534)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4026..4045)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4625..5413)
                     /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      5871..6306
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 tggtggcagg atatattgtg gtgtaaacaa attgacgctt agacaactta ataacacatt
       61 gcggacgttt ttaatgtaat cagcttgcat gccggtcgat ctagtaacat agatgacacc
      121 gcgcgcgata atttatccta gtttgcgcgc tatattttgt tttctatcgc gtattaaatg
      181 tataattgcg ggactctaat cataaaaacc catctcataa ataacgtcat gcattacatg
      241 ttaattatta catgcttaac gtaattcaac agaaattata tgataatcat cgcaagaccg
      301 gcaacaggat tcaatcttaa gaaactttat tgccaaatgt ttgaacgatc tgcttgactc
      361 taggggtcat cagatttcgg tgacgggcag gaccggacgg ggcggcaccg gcaggctgaa
      421 gtccagctgc cagaaaccca cgtcatgcca gttcccgtgc ttgaagccgg ccgcccgcag
      481 catgccgcgg ggggcatatc cgagcgcctc gtgcatgcgc acgctcgggt cgttgggcag
      541 cccgatgaca gcgaccacgc tcttgaagcc ctgtgcctcc agggacttca gcaggtgggt
      601 gtagagcgtg gagcccagtc ccgtccgctg gtggcggggg gacacgtaca cggtcgactc
      661 ggccgtccag tcgtaggcgt tgcgtgcctt ccagggaccc gcgtaggcga tgccggcgac
      721 ctcgccgtcc acctcggcga cgagccaggg atagcgctcc cgcagacgga cgaggtcgtc
      781 cgtccactcc tgcggttcct gcggctcggt acggaagttg accgtgcttg tctcgatgta
      841 gtggttgacg atggtgcaga ccgccggcat gtccgcctcg gtggcacggc ggatgtcggc
      901 cgggcgtcgt tctgggctca tggtagatcc cctcgatcga gttgagagtg aatatgagac
      961 tctaattgga taccgagggg aatttatgga acgtcagtgg agcatttttg acaagaaata
     1021 tttgctagct gatagtgacc ttaggcgact tttgaacgcg caataatggt ttctgacgta
     1081 tgtgcttagc tcattaaact ccagaaaccc gcggctcagt ggctccttca acgttgcggt
     1141 tctgtcagtt ccaaacgtaa aacggcttgt cccgcgtcat cggcgggggt cataacgtga
     1201 ctcccttaat tctccgctca tgatcgataa catgtcgact atcggacgag ctcaaagttt
     1261 aacgtgtgaa gacgtgcttc taatgtagtt taaactgaag gcgggaaacg acaatctgat
     1321 ccaagctcaa gctaagcttg catgcctgca ggatatcgtg gatccaagct tgccacgtgc
     1381 cgccacgtgc cgccacgtgc cgccacgtgc ctctagagga tccatctcca ctgacgtaag
     1441 ggatgacgca caatcccact atccttcgca agacccttcc tctatataag gaagttcatt
     1501 tcatttggag aggacacgct gggatcccca ccatggctgg cgccgtgggc agacccagaa
     1561 gatctgctcc tcggagagcc ggcaagaacc cccgggaaga gattctggat gccagcgccg
     1621 agctgttcac cagacagggc tttgccacca ccagcaccca ccagattgcc gacgctgtgg
     1681 gcatcagaca ggccagcctg tactaccact tccccagcaa gaccgagatc ttcctgaccc
     1741 tgctgaaaag caccgtggaa ccctccaccg tgctggccga ggatctgtct accctggacg
     1801 ccggacccga aatgagactg tgggctatcg tggccagcga agtgcggctg ctgctgagca
     1861 ccaagtggaa cgtgggccgg ctgtaccagc tgcccatcgt gggctctgag gaattcgccg
     1921 agtaccacag ccagcgcgag gccctgacca acgtgttcag agatctggcc accgagattg
     1981 tgggcgacga ccccagagcc gaactgccct tccacatcac catgagcgtg atcgagatgc
     2041 ggcggaacga cggcaagatc cctagccctc tgagcgccga ctctctgccc gagacagcca
     2101 ttatgctggc tgacgcctcc ctggctgtgc tgggagcacc tctgcctgcc gacagagtgg
     2161 aaaagacact ggaactgatc aagcaggccg acgccaaggc cctttcttct cttagtagag
     2221 agctcgtttt cttgatctta cagtttctcg atgaagagaa gtttaaagag actgttcaca
     2281 agcttgaaca agaatctggg tttttcttca atatgaagta ttttgaggat gaggttcaca
     2341 atggaaattg ggatgaggtt gagaagtatc tctctggttt cactaaagta gatgataata
     2401 gatactctat gaagatattc ttcgagataa gaaagcagaa gtatctcgag gctttggata
     2461 agcatgatcg tcccaaggct gtggatatac tagtgaagga tttgaaagtg ttttcaactt
     2521 ttaatgagga gcttttcaag gaaataacac agctcttgac attggagaac ttccgggaga
     2581 atgaacagtt atccaagtat ggggacacca agtctgcaag agcgatcatg ttggtggaac
     2641 tcaaaaagct gattgaggcg aatcctttat tccgtgataa attgcagttc cctactctta
     2701 gaaattcaag gctgaggact ttgatcaacc agagcttaaa ttggcaacac cagctttgta
     2761 aaaatccaag gccaaatcca gatatcaaga ctctttttgt ggatcactct tgtggccctc
     2821 caaatggcgc acgagctcca tctcctgtga acaatccact gcttggaggg ataccaaagg
     2881 ctggaggatt tcctccttta ggcgcacacg ggccatttca accaacggcg tcacctgttc
     2941 caacacctct tgctggttgg atgtctagtc catcctctgt cccacatcca gctgtctcag
     3001 caggagccat tgcacttggt ggtccatcaa tcccagcggc gttgaagcac ccgagaactc
     3061 ctccaactaa tgcttcttta gactacccgt cagcagattc tgaaccgaag aagaagagga
     3121 aggtttagtg tcggtacgct gaaatcacca gtctctctct acaaatctat ctctctctat
     3181 tttctccata aataatgtgt gagtagtttc ccgataaggg aaattagggt tcttataggg
     3241 tttcgctcat gtgttgagca tataagaaac ccttagtatg tatttgtatt tgtaaaatac
     3301 ttctatcaat aaaatttcta attcctaaaa ccaaaatcca gtactaaaat ccagatcgat
     3361 gcttgatact acgtcttcgg gctagagcgg actagttcta gagcggccgc caccgcggtg
     3421 gagctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc
     3481 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     3541 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     3601 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     3661 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     3721 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3781 acggttatcc acagaatcag gggataacgc aggaaagaac atgaaggcct tgacaggata
     3841 tattggcggg taaactaagt cgctgtatgt gtttgtttga gatctcatgt gagcaaaagg
     3901 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3961 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4021 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4081 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4141 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4201 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4261 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4321 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4381 tagaagaaca gtatttggta tctgtgctct gctgaagcca gttaccttcg gaagaagagt
     4441 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4501 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4561 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga attcttgtta
     4621 gacattattt gccgactacc ttggtgatct cgcctttcac gtagtgaaca aattcttcca
     4681 actgatctgc gcgcgaggcc aagcgatctt cttgtccaag ataagcctgc ctagcttcaa
     4741 gtatgacggg ctgatactgg gccggcaggc gctccattgc ccagtcggca gcgacatcct
     4801 tcggcgcgat tttgccggtt actgcgctgt accaaatgcg ggacaacgta agcactacat
     4861 ttcgctcatc gccagcccag tcgggcggcg agttccatag cgttaaggtt tcatttagcg
     4921 cctcaaatag atcctgttca ggaaccggat caaagagttc ctccgccgct ggacctacca
     4981 aggcaacgct atgttctctt gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg
     5041 ctggctcgaa gatacctgca agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc
     5101 gcttagctgg ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg acttctacag
     5161 cgcggagaat ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag
     5221 ctcgccgcgt tgtttcatca agccttacgg tcaccgtaac cagcaaatca atatcactgt
     5281 gtggcttcag gccgccatcc actgcggagc cgtacaaatg tacggccagc aacgtcggtt
     5341 cgagatggcg ctcgatgacg ccaactacct ctgatagttg agtcgatact tcggcgatca
     5401 ccgcttccct catgatgttt aactcctgaa ttaagccgcg ccgcgaagcg gtgtcggctt
     5461 gaatgaattg ttaggcgtca tcctgtgctc ccgagaacca gtaccagtac atcgctgttt
     5521 cgttcgagac ttgaggtcta gttttatacg tgaacaggtc aatgccgccg agagtaaagc
     5581 cacattttgc gtacaaattg caggcaggta cattgttcgt ttgtgccatg atgatatatt
     5641 tttatcttgt gcaatgtaac atcagagatt ttgagacaca acgtggcttt gttgaataaa
     5701 tcgaactttt gctgagttga aggatcagat cacgcatctt cccgacaacg cagaccgttc
     5761 cgtggcaaag caaaagttca aaatcaccaa ctggtccacc tacaacaaag ctctcatcaa
     5821 ccgtggctcc ctcactttct ggctggatga tggggcgatt caggagatcc ccatccaaca
     5881 gcccgccgtc gagcgggctt ttttatcccc ggaagcctgt ggatagaggg tagttatcca
     5941 cgtgaaaccg ctaatgcccc gcaaagcctt gattcacggg gctttccggc ccgctccaaa
     6001 aactatccac gtgaaatcgc taatcagggt acgtgaaatc gctaatcgga gtacgtgaaa
     6061 tcgctaataa ggtcacgtga aatcgctaat caaaaaggca cgtgagaacg ctaatagccc
     6121 tttcagatca acagcttgca aacacccctc gctccggcaa gtagttacag caagtagtat
     6181 gttcaattag cttttcaatt atgaatatat atatcaatta ttggtcgccc ttggcttgtg
     6241 gacaatgcgc tacgcgcacc ggctccgccc gtggacaacc gcaagcggtt gcccaccgtc
     6301 gagcgccttt gcccacaacc cggcggccgg ccgcaacaga tcgttttata aatttttttt
     6361 tttgaaaaag aaaaagcccg aaaggcggca acctctcggg cttctggatt tccgatcccc
     6421 ggaattagac tctggctgcc tgtatcgagt ggtgattttg tgccgagctg ccggtcgggg
     6481 agctgttggc tggc
//
