LOCUS       sequence-395624-        4756 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains RALF23 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4756)
  AUTHORS   Calabria J, Wang L, Rast-Somssich MI, Chen HW, Watt M, Persson S, 
            Andersen TG, Idnurm A, Somssich M
  TITLE     Resolving spatially distinct phytohormone response zones in 
            Arabidopsis thaliana roots colonized by Fusarium oxysporum.
  JOURNAL   J Exp Bot. 2025 Jan 17:erae516. doi: 10.1093/jxb/erae516.
  PUBMED    39823275
REFERENCE   2  (bases 1 to 4756)
  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"; journalName: "J Exp Bot. 
            2025 Jan 17:erae516. doi: 10.1093/jxb/erae516."
COMMENT     Sequence Label: pGG-PIP04
FEATURES             Location/Qualifiers
     source          1..4756
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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        2212..2230
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2212..2229
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4342..4361)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4343..4361)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4371..4393)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4587..4606)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4706..4728
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtggctct cgaggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacgagc tatttaggtg
     2221 acactataga agtgaagctt ggtctcaacc tggtgattcc ggtttccgac ggattcgtcg
     2281 cggtcaacag tcaacactaa agtaccaaac tattcaaaac ttttacgttt ctacgtgtat
     2341 ttgtcaagtt gactaaaagc cgcttctcca gatattttac gtcttgccat ttattgttgg
     2401 gccttgattg tttgggcctt tcaaatatta tatttctttt tttttgagtt ttaatagggg
     2461 gcccgtttta atagtctatt tggactttct ttttctctgt actggacaaa aagggtatta
     2521 cttttatatt tttcattact aaatttcagt agatttaata tctttggttg agaacaaaat
     2581 tgttgtataa ataataaatt acttgtatct tgtattttga agagtcattg aaatttatac
     2641 tccaaatctt agtattcata agaaggaaac gagtaaatcg gtgtgaacct aaaattttgt
     2701 agtcacacct aaaaacctaa aattgcaacg atttaaaaac tataaaacct tcgcatataa
     2761 atccaaataa aatgtcttaa gaaaatgagt tatttttttt gaacatttag ctaataaaat
     2821 tttcttcaac atatgctata caaattatat attacttttt taaagaatag tttttttaaa
     2881 aacgatttac atatctcttt ttgtatattg tagatcatga ataatatgac cattgatttt
     2941 ctttattttt ttagaagatg actttataat acttaaaact tttaagatct ttttaaatat
     3001 aatttcacaa aatttacaat gtatttatat attttattgt agatcatgaa taacataacc
     3061 aatcttgcgt tttttcttga ctgtagaatg accatctctg aaatttcagg caatggtctg
     3121 agatatagat ttccagcctc cttgtgtgaa cactcaagca gaccattaca ctcaacacat
     3181 cagctactaa catttgttga aatttcatgt acatataaat gattcttggc gccttgattc
     3241 atcaacattt ttaaatataa acttctttcc ctttctcaat atcgattcca atttcaggat
     3301 tcgaaccttc gaaaacatcg taatggagaa cctacgattt caatatgaaa aacacacaca
     3361 catcattcaa taattcagaa caacttagaa accatcaaat tgcaaacccg cttcggatca
     3421 tgtcttagga aaaaaataat ccaaatagtg tccactctca tttcataatt gttgctttgt
     3481 gtcgttatgc ctgtcatgat atggttttta ataatgtcca ttttgatttt ctttttggtt
     3541 tttataaatt agttacaaac ttataatgtc cattttgatg aaattggaaa tcacaaatat
     3601 gaacaataca caagagatgt acattaaact cccttcatta atatatgttg gcattttaat
     3661 aggtagtttt ttttttcatt tgaaatagtc tcaaatcatg gaaggagagc actcataatt
     3721 gtacaaaata aaagtaaatg cgttctatat tactcttaag tcttatctac ctttgtacta
     3781 tcgttttaat ttcttgttag tatagtagta ccttgaaaca taagcaaaaa cataaacaga
     3841 actttttatt aatttttttt ttggtagaaa agaacactgt atttctactt acgattctaa
     3901 taaattataa caatttaaaa cagtatactt tccaaaataa attagcaaaa tgtaaattat
     3961 tcatttatat aacaaaagaa attttactaa tttgtaaaat tgtgtggatg gttggtgaat
     4021 atgattttag tatatcgtaa aagtgaccaa caaaatcttt aagaaaaaaa agaaaaataa
     4081 gtagtggatg tctccatcaa aaacacttgg cgcattaaaa catccttaga tctgaagcta
     4141 acaaaaggcc acttgtaaag ctgtctcttt acgacaccaa gtggtattat tcgtaattac
     4201 ttccatcttt cagtggcatt tcattaagct ccttctcttt taaaaacctt ttcttttatc
     4261 tctctatctc tttcttcata gttcgtgcac agagagagct aaagctacag tgtacagaag
     4321 aaacatgaga ccgaattctc gccctatagt gagtcgtatt acctggccgt cgttttacaa
     4381 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     4441 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     4501 agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     4561 tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     4621 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     4681 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     4741 tcaccgaaac gcgcga
//
