LOCUS       sequence-112032-        7490 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The gRNA construct is designed for constitutive expression of 
            salicylate hydroxylase for degradation of salicylic acid to 
            catechol..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7490)
  AUTHORS   Todesco M, Balasubramanian S, Hu TT, Traw MB, Horton M, Epple P, 
            Kuhns C, Sureshkumar S, Schwartz C, Lanz C, Laitinen RA, Huang Y, 
            Chory J, Lipka V, Borevitz JO, Dangl JL, Bergelson J, Nordborg M, 
            Weigel D
  TITLE     Natural allelic variation underlying a major fitness trade-off in 
            Arabidopsis thaliana.
  JOURNAL   Nature. 2010 Jun 3;465(7298):632-6. doi: 10.1038/nature09083.
  PUBMED    20520716
REFERENCE   2  (bases 1 to 7490)
  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"; journalName: 
            "Nature"; date: "2010-06-3- 3"; volume: "465"; issue: "7298"; pages:
            "632-6"
COMMENT     Sequence Label: MT363
FEATURES             Location/Qualifiers
     source          1..7490
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    542..564
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     terminator      complement(588..839)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(859..1410)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIQTSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     promoter        complement(1453..1632)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     1864..1886
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1878..1895
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1879..1895
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1905..1924
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1905..1923
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2454..2799
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2742..2762
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     2863..2879
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2864..2880
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4268..4292)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4300..4316)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4300..4316)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      4320..5027
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(5060..5078)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5061..5078)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5117..5137)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5117..5135)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5156..5172)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5156..5172)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5169..5191)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5180..5196
                     /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(5204..5210,5211..5228,5229..5234))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5249..5270
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5387..5404)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5509..5533
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5624..6212)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5704..5723)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6383..7171)
                     /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      join(7462..7490,1..407)
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 tttttatccc cggaagcctg tggatagagg gtagttatcc acgtgaaacc gctaatgccc
       61 cgcaaagcct tgattcacgg ggctttccgg cccgctccaa aaactatcca cgtgaaatcg
      121 ctaatcaggg tacgtgaaat cgctaatcgg agtacgtgaa atcgctaata aggtcacgtg
      181 aaatcgctaa tcaaaaaggc acgtgagaac gctaatagcc ctttcagatc aacagcttgc
      241 aaacacccct cgctccggca agtagttaca gcaagtagta tgttcaatta gcttttcaat
      301 tatgaatata tatatcaatt attggtcgcc cttggcttgt ggacaatgcg ctacgcgcac
      361 cggctccgcc cgtggacaac cgcaagcggt tgcccaccgt cgagcgccag cgcctttgcc
      421 cacaacccgg cggccggccg caacagatcg ttttataaat tttttttttt gaaaaagaaa
      481 aagcccgaaa ggcggcaacc tctcgggctt ctggatttcc gatccccgga attagagatc
      541 ttggcaggat atattgtggt gtaacgttat cagcttgcat gccggtcgat ctagtaacat
      601 agatgacacc gcgcgcgata atttatccta gtttgcgcgc tatattttgt tttctatcgc
      661 gtattaaatg tataattgcg ggactctaat caaaaaaccc atctcataaa taacgtcatg
      721 cattacatgt taattattac atgcttaacg taattcaaca gaaattatat gataatcatc
      781 gcaagaccgg caacaggatt caatcttaag aaactttatt gccaaatgtt tgaacgatct
      841 gcttgactct aggggtcatc agatttcggt gacgggcagg accggacggg gcggcaccgg
      901 caggctgaag tccagctgcc agaaacccac gtcatgccag ttcccgtgct tgaagccggc
      961 cgcccgcagc atgccacggg gggcatatcc gagcgcctcg tgcatgcgca cgctcgggtc
     1021 gttgggcagc ccgatgacag cgaccacgct cttgaagccc tgtgcctcca gggacttcag
     1081 caggtgggtg tagagcgtgg agcccagtcc cgtccgctgg tggcgggggg agacgtacac
     1141 ggttgactcg gccgtccagt cgtaggcgtt gcgtgccttc cagggacccg cgtaggcgat
     1201 gccggcgacc tcgccgtcca cctcggcgac gagccaggga tagcgctccc gcagacggac
     1261 gaggtcgtcc gtccactcct gcggttcctg cggctcggta cggaagttga ccgtgcttgt
     1321 ctggatgtag tggttgacga tggtgcagac cgccggcatg tccgcctcgg tggcacggcg
     1381 gatgtcggcc gggcgtcgtt ctgggctcat ggtagatccc cctcgatcga gttgagagtg
     1441 aatatgagac tctaattgga taccgagggg aatttatgga acgtcagtgg agcatttttg
     1501 acaagaaata tttgctagct gatagtgacc ttaggcgact tttgaacgcg caataatggt
     1561 ttctgacgta tgtgcttagc tcattaaact ccagaaaccc gcggctcagt ggctccttca
     1621 acgttgcggt tctgtcagtt ccaaacgtaa aacggcttgt cccgcgtcat cggcgggggt
     1681 cataacgtga ctcccttaat tctccgctca tgatcgataa cattaacgct tacaatttcc
     1741 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     1801 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     1861 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat
     1921 agggcgaatt gggtaccggg ccccccctcg atatgattcg gtccccagat tagccttttc
     1981 aatttcagaa agaatgctaa cccacagatg gttagagagg cttacgcagc aggtctcatc
     2041 aagacgatct acccgagcaa taatctccag gaaatcaaat accttcccaa gaaggttaaa
     2101 gatgcagtca aaagattcag gactaactgc atcaagaaca cagagaaaga tatatttctc
     2161 aagatcagaa gtactattcc agtatggacg attcaaggct tgcttcacaa accaaggcaa
     2221 gtaatagaga ttggagtctc taaaaaggta gttcccactg aatcaaaggc catggagtca
     2281 aagattcaaa tagaggacct aacagaactc gccgtaaaga ctggcgaaca gttcatacag
     2341 agtctcttac gactcaatga caagaagaaa atcttcgtca acatggtgga gcacgacaca
     2401 cttgtctact ccaaaaatat caaagataca gtctcagaag accaaagggc aattgagact
     2461 tttcaacaaa gggtaatatc cggaaacctc ctcggattcc attgcccagc tatctgtcac
     2521 tttattgtga agatagtgga aaaggaaggt ggctcctaca aatgccatca ttgcgataaa
     2581 ggaaaggcca tcgttgaaga tgcctctgcc gacagtggtc ccaaagatgg acccccaccc
     2641 acgaggagca tcgtggaaaa agaagacgtt ccaaccacgt cttcaaagca agtggattga
     2701 tgtgatatct ccactgacgt aagggatgac gcacaatccc actatccttc gcaagaccct
     2761 tcctctatat aaggaagttc atttcatttg gagagaacac gggggacgag ctcgcccatc
     2821 acaagtttgt acgtacaaaa aagcaggctt tcgggccccc cctcgaggtc gacggtatcg
     2881 ataagcttga tatcgaattc ctgcagcccg ggggatccac tagtgattga attcatgaaa
     2941 aacaataaac ttggcttgcg catcggtatc gtcggcggcg gaatttccgg cgttgcctta
     3001 gcactggaac tctgccgcta ctcccatatc caggtacagc tgttcgaggc tgcgccggct
     3061 ttcggtgagg tcggtgccgg cgtgtccttt ggccccaacg cggtgcgcgc cattgtcggc
     3121 ctgggcttgg gcgaggccta cctgcaggtt gccgaccgta cttcggagcc ctgggaggac
     3181 gtgtggttcg aatggcggcg cggcagcgat gccagctatc tgggagccac catcgctccg
     3241 ggcgtgggcc agtcctcggt acaccgggcg gatttcatcg acgccctagt aactcacctc
     3301 ccagaaggta tcgcccaatt cgggaagcgc gccacccagg tcgagcagca gggtggcgaa
     3361 gtgcaagtgc tgttcaccga cggcacagag taccgctgcg accttctgat cggtgccgac
     3421 ggaatcaagt cagcgctccg tagccatgtg ctggaaggtc aggggctggc cccacaagtg
     3481 ccgcgattca gcggcacctg tgcctatcgg gggatggtcg acagcctgca tctgcgagaa
     3541 gcctatcggg cccatggcat cgacgagcac ttggtggacg tgccgcagat gtacctaggg
     3601 ctcgacggcc atatcctcac ctttccagtg aggaatggcg gcatcatcaa cgtggtggct
     3661 ttcatctccg accgtagcga gccgaagccg acctggcctg cggatgcccc ttgggtgcgt
     3721 gaggcgagcc agcgcgagat gctcgatgcc ttcgcgggtt ggggggatgc cgcgcgcgcc
     3781 ctgctggagt gcatcccggc accaactctc tgggcactgc atgacctggc ggagctgccg
     3841 ggctacgtgc acggtcgggt cgtcctgatc ggcgacgcag ctcacgccat gctgccgcac
     3901 caaggtgccg gtgctggcca agggcttgag gacgcctact tcctcgcccg cctgttgggc
     3961 gatacgcagg ccgatgccgg caacctcgcc gagctgcttg aagcctacga cgacctgcgc
     4021 cgccctcgtg cctgtcgcgt gcagcaaacc tcctgggaga ccggcgagtt atacgagttg
     4081 cgcgaccccg tcgtcggtgc gaacgagcag ctgctggggg aaaacctggc gacccgcttc
     4141 gactggctgt ggaaccacga cctcgacact gacctggccg aggcccgtgc gcggctgggt
     4201 tgggagcatg gtggcggggg tgtgctacgt caagggctcg agaatcgaat tcccgcggtg
     4261 gatctagacc cagctttctt gtacaaagtg gtgatggggg atccactagt tctagagtcc
     4321 tgctttaatg agatatgcga gacgcctatg atcgcatgat atttgctttc aattctgttg
     4381 tgcacgttgt aaaaaacctg agcatgtgta gctcagatcc ttaccgccgg tttcggttca
     4441 ttctaatgaa tatatcaccc gttactatcg tatttttatg aataatattc tccgttcaat
     4501 ttactgattg taccctacta cttatatgta caatattaaa atgaaaacaa tatattgtgc
     4561 tgaataggtt tatagcgaca tctatgatag agcgccacaa taacaaacaa ttgcgtttta
     4621 ttattacaaa tccaatttta aaaaaagcgg cagaaccggt caaacctaaa agactgatta
     4681 cataaatctt attcaaattt caaaagtgcc ccaggggcta gtatctacga cacaccgagc
     4741 ggcgaactaa taacgctcac tgaagggaac tccggttccc cgccggcgcg catgggtgag
     4801 attccttgaa gttgagtatt ggccgtccgc tctaccgaaa gttacgggca ccattcaacc
     4861 cggtccagca cggcggccgg gtaaccgact tgctgccccg agaattatgc agcatttttt
     4921 tggtgtatgt gggccccaaa tgaagtgcag gtcaaacctt gacagtgacg acaaatcgtt
     4981 gggcgggtcc agggcgaatt ttgcgacaac atgtcgaggc tcagcaggac cggcatgcaa
     5041 gctagcttac tagtgatatt ctatagtgtc acctaaatct gcggccgcca ccgcggtgga
     5101 gctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat
     5161 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5221 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5281 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5341 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5401 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5461 ggttatccac agaatcaggg gataacgcag gaaagaacat gaaggccttg acaggatata
     5521 ttggcgggta aactaagtcg ctgtatgtgt ttgtttgaga tctcatgtga gcaaaaggcc
     5581 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5641 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5701 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5761 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5821 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5881 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5941 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6001 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6061 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga agaagagttg
     6121 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     6181 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6241 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6301 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6361 atgtgtaaca ttggtctagt gattatttgc cgactacctt ggtgatctcg cctttcacgt
     6421 agtgaacaaa ttcttccaac tgatctgcgc gcgaggccaa gcgatcttct tgtccaagat
     6481 aagcctgcct agcttcaagt atgacgggct gatactgggc cggcaggcgc tccattgccc
     6541 agtcggcagc gacatccttc ggcgcgattt tgccggttac tgcgctgtac caaatgcggg
     6601 acaacgtaag cactacattt cgctcatcgc cagcccagtc gggcggcgag ttccatagcg
     6661 ttaaggtttc atttagcgcc tcaaatagat cctgttcagg aaccggatca aagagttcct
     6721 ccgccgctgg acctaccaag gcaacgctat gttctcttgc ttttgtcagc aagatagcca
     6781 gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag aatgtcattg cgctgccatt
     6841 ctccaaattg cagttcgcgc ttagctggat aacgccacgg aatgatgtcg tcgtgcacaa
     6901 caatggtgac ttctacagcg cggagaatct cgctctctcc aggggaagcc gaagtttcca
     6961 aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag ccttacggtc accgtaacca
     7021 gcaaatcaat atcactgtgt ggcttcaggc cgccatccac tgcggagccg tacaaatgta
     7081 cggccagcaa cgtcggttcg agatggcgct cgatgacgcc aactacctct gatagttgag
     7141 tcgatacttc ggcgatcacc gcttccctca taacacccct tgtattactg tttatgtaag
     7201 cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat
     7261 tttgagacac aacgtggctt tgttgaataa atcgaacttt tgctgagttg aaggatcaga
     7321 tcacgcatct tcccgacaac gcagaccgtt ccgtggcaaa gcaaaagttc aaaatcacca
     7381 actggtccac ctacaacaaa gctctcatca accgtggctc cctcactttc tggctggatg
     7441 atggggcgat tcaggcgatc cccatccaac agcccgccgt cgagcgggct
//
