LOCUS       sequence-396370-        4380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains PEP3 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4380)
  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 4380)
  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-PIP03
FEATURES             Location/Qualifiers
     source          1..4380
                     /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(3966..3985)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3967..3985)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3995..4017)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4211..4230)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4330..4352
                     /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 tgcactttaa gttacattgt ttagtctaat
     2281 tatttgcagt cgaaatgtgt taatttaata tcactgtttt acttttttat tatatcaaca
     2341 atctacagac aaacaaaatt tcattaagtt cttgttcact atacgagttt tttcctcttt
     2401 atttcatgca tttttttttt cccgtatttg atgtgatatt tgggttgaac aaaagtaaat
     2461 atgttcgacc atcataagtt cataaccagt atcaatatta attgaacgca tttaatatcc
     2521 atccatatcg aacgtatctg tcaaatgtta aacaacatta aagacgaacc taattgatat
     2581 ttttggattt attttaacag tcaacagcta tttggttcta cctagctagc tcacattaat
     2641 tataatttaa tgtcttcact catagaatgg tcatgtaaaa ttactaaaga attaagtttt
     2701 gcattcacca tacttttgtt tatagatcat cataccaata atttataaag ttagatgtta
     2761 taaaataaaa taaacaaaaa ccttaaacgc agtgataatc atcctatcac aattcacaac
     2821 gtgcattatg aatggtaata ttatatacaa agttgacgag tgctctctat gagtaacgca
     2881 tgatcgctct atgatttaat aaaatatact ccacgaggag cagaagcaac accaacttcg
     2941 aatataaaac acatttaaca atcttcactg tgactttaga catcatgatt tttttggtgt
     3001 tcttttttta aaaaactcat aatctggtga taagtctcgt agttacctca tcgtagaacc
     3061 acaatttagt atttacacaa gaaggtatgg tctgattaaa gtttgggggt ctttgtaatt
     3121 aggcctacaa taatcattca caaatttgaa atattacaat ttatgaggga ccacaacacc
     3181 taaaaaggca taaatgcatt atgcccagtg gcgacgaaaa aagtgcatta aaccaaagga
     3241 atatatgttt gatttttatt ttaatgtggg aatgggatat gcaatattac aattatgata
     3301 ttatctgaaa ttttatatct ttacaaaatt tgattttcaa acttaaattc tatagtatga
     3361 tctacatgaa cactttatat tcttatgcaa gaaaaacaag tcctttgtcc aatcattgac
     3421 ctcttaattg aatttgaaat atttaacaaa gtacccaata gataattctt ttgttcttgt
     3481 taaattagat tttcaaaata ctaagcttta gcattaattt gaataatcaa agatcttcct
     3541 ttgaattacc tattcaacat ttgtttaata acttaattca aacaaaaacc aaagaagagc
     3601 gtgaattgac tttgaccaaa accaaattct ctcaagagtt gctgccatat tttgaccgcg
     3661 cgccacgtag attagagtca aaattgttat tttattcact tcaacataaa accaaataag
     3721 cattatcggt tttcaacata ccggcaccaa cttttccaaa gtctgtatgt acctaacaaa
     3781 accggtttat catagaaacg gtcaacacac caaaaatagt tgaccaacaa ctacccaagt
     3841 gatatccctt taaaaggagt cgcatatgtg ttaccaagtt ccatcatcaa cctaataaca
     3901 cacaacacta aatctctttc ccaaaaaaag attaagaagt caacgaacat gagaccgaat
     3961 tctcgcccta tagtgagtcg tattacctgg ccgtcgtttt acaacgtcgt gactgggaaa
     4021 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     4081 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     4141 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt
     4201 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     4261 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     4321 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
//
