LOCUS       sequence-396363-        6034 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains TUB6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6034)
  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 6034)
  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-PIP75
FEATURES             Location/Qualifiers
     source          1..6034
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(13..30)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(184..772)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(264..283)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(943..1734,1735..1803))
                     /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     1566..1585
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1804..1908)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1976..1994
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2032..2054)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2154..2173
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2367..2389
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2399..2418
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2399..2417
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(5808..5826)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5809..5826)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    5840..5856
                     /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(5864..5870,5871..5888,5889..5894))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5909..5930
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
       61 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
      121 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
      181 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      241 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      301 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      361 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      421 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      481 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      541 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      601 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
      661 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      721 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
      781 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
      841 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
      901 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
      961 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     1021 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     1081 tcgagagcca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     1141 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     1201 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     1261 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     1321 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     1381 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     1441 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     1501 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     1561 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     1621 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     1681 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     1741 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     1801 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     1861 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     1921 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     1981 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     2041 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     2101 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     2161 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     2221 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     2281 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     2341 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccaggta
     2401 atacgactca ctatagggcg agaattcggt ctcatgttct tctattttat ctgaaatcaa
     2461 cattacatga acaaaacagt aaacaacgga ttttgaaagc aaaagaatct gtatcagatc
     2521 caaatgaaat tatatcgaat gtcaacaact aaatctcgaa gatctgaaat caaacaatct
     2581 gtatcagatc ccaaaacaaa tcaatcggaa aaaaacacaa cgccgcagag aagaaaactc
     2641 cgatttcaaa ttcttcacgg aattcaaaac ggcaacacat tttcaagtga tagaaaatca
     2701 aaatccaagt taagagagaa aatatgaaac agatcaagag cggatacaga tttggaggaa
     2761 gacgattcga tcaaagtatc acaataagta gtaggagttc ataatctcat tccctagaga
     2821 ggaaacaata cataaaaagg aaatagcaga gtaaacaata actcacggtg cagaggagaa
     2881 tcccggcgag attgggataa tcgtcgaccg gagtaggaga ggaagtgaga cgggtggaga
     2941 gtgaagaagt tggatttatt tatgaaagag taagaagagg gaaaggtaaa atttgatcca
     3001 acggctgtga ttggtttaat ctatcggaag ctggatgccg ttggatggtg agatatcaac
     3061 gttgaggatg gcgttgggta agatgtggga gctactttat gttaattgga ttgtgattgt
     3121 cggttcttcc ttcactccta cataccaatt attgttgtca catgaatagt aaaaaaatct
     3181 ctaacattgg agtcttggac cattggtcta acatagaaaa gataagtatt tcggtagtaa
     3241 tgtcaagatt cttattattt tccaaacgca acttacaatt ttagagaagc tctctttggt
     3301 tacacactta atacacgtca aatgcataaa caaactcatc aaaagccgta ggaacaggca
     3361 ggagcaatac cttgtaatga gtgtgtgaca gtataaaatc gagctaagaa ccacgggaaa
     3421 aaaaaatggc ctcattgcgt atgacaatgc ggagaatagt aaatcaccgt accatggtcg
     3481 gagagtagat gaattcttaa tggtttattt cgtacctata ttaactagtt agttaagttt
     3541 agttactttc gtaaaatata tattaaatct ttcttttgca tgcatgtcat agcaatcagt
     3601 ttatatgaaa tatatgcaga aattgttttt tggtccatca gatctatacg agagagttga
     3661 agaaaaaagt tgatgcatat taccgtagtt aggacgataa tgtaaaacac acgcattgga
     3721 tactctcgtt tagatttaac tactgatgct aaattgctac tttgaaatct ttaaaatatt
     3781 ctataagttc ttacatctgt cgatctactg tcaatcaata aatatatcag tttgctattc
     3841 ggaaaatcta aactaataat cggataactt aaatattgtt ttctcaactt tgtcgttgct
     3901 tgtataacaa taaaaaagtg atgtttcgat ggtttcgctt agtactaaat tttccatgga
     3961 acattaatca tgcttacgtt tctactttct ataacaatgc aaaagaaaac ccgaaaccaa
     4021 aagcactact aatgatatca ataacaaaac caatgtcgcc cttatatcac catcatttag
     4081 atcataaaca agacgtgata aataaattac gctaagaaca gcatatagct gtaaatatga
     4141 gaaattagaa gggtacaaaa acaagaatgt gttggtcttg tagagcttgt ggatcggaaa
     4201 tcggatagcg acagaatgag atgtagagat atataaaaag aacaatgcac atgaaatcca
     4261 cattgcgtcc catgtgacat gccaacagct cacctgactt aaattctaac tttctctact
     4321 cgtagtttct tgctccagta gtttctcata cctacaaagt tttttgctaa attccaatcc
     4381 ctacctacaa agtttgtcag agttaatgtc gttataatat cttcttaatt tatttttttc
     4441 ccacaagttc ttgtaaatac aaaaattgaa tatcttgtag tataagtata tatcaagtga
     4501 agtttactct ttttgtaagt agtagtatta tttacgtgat taatattata atttgtttct
     4561 atgcaataag tcttgactct ttcaaataaa tcctctcact tgatctgtac gaattttctg
     4621 gtatttctat gtctgtgaat attcatactt tttgttatgg ttaaccgagg atgagcgatg
     4681 tccaaaagag attagttgag tgctctttga tgatgaaact ctatgaaata gaaacacatc
     4741 tctctttgaa gattattgtt caaagcaaac gaagtgattg gtcaatgcag ctcttgagaa
     4801 ttcaacaaga gcaacaacgt aatcaattag ctgaagtagt gaagtctatc attttctttt
     4861 gctatgatca aaattgtaat tcattaattt attggtttgt gattgtgaaa aagttttaat
     4921 gacttttaaa atatataaaa aatagataaa ttaaaagatt ttaaatgatt actagagagt
     4981 tttgtataat ttttatgatt tgtttttcta aaattttgta aagtctttct ataatcttgt
     5041 aaaatctcct tgaatataat tataaacttt aatttgtgat tttttcaaaa ttataacata
     5101 tctccaaaat atagtatact attcttagtt attttcatca aaatgaaaat cacacgcata
     5161 aacacaataa ctatggtatt tgtaagtaaa gttacatgaa tagggaacca tccatgaaga
     5221 acttgatgtt ttattttcaa atattttgtt ggatatttac ttttcatatg ttagattaaa
     5281 atgctttgtt ttaaaaattg cttgtgtgtt gatttttcac aaagagatta tagaggagca
     5341 ttcgtgggac ttgacccatt agaatatata tattgttcat cactgttaat gagatttatg
     5401 catctgttcg tatcattggt ttttgtttat atccaaaatc atgattcaca atctctcttt
     5461 ttttatgata cttttcatga ttttaatttg tgaagaaaat aagataaaat cataatcgaa
     5521 taacacaata ttttataatc attaacagtc atggaaattg ttgtttaagt tgaataacac
     5581 aaaatttgta atgattttat taaagtatca atctaataac tccagattta caaaaacttt
     5641 aaatttcttt tttacaaatc aaaatctaat aacaccaaac tttaaaaaaa tcttacaaaa
     5701 ttttaattaa ataacaaaaa aaaattcatg acattacaaa tcatttaaaa tctatataaa
     5761 tcacaaacca ataacaccct ctaaataggt tgagaccaag cttcacttct atagtgtcac
     5821 ctaaatagct cgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5881 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5941 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     6001 aacgcgcggg gagaggcggt ttgcgtattg ggcg
//
