LOCUS       sequence-395622-        4754 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains TET8 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4754)
  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 4754)
  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-PIP66
FEATURES             Location/Qualifiers
     source          1..4754
                     /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(4340..4359)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4341..4359)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4369..4391)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4585..4604)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4704..4726
                     /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 tcggatgtat caaaggtaaa aatatcaaaa
     2281 atgtgtcagt tttcaaattt aggacgaaaa aacttaaact ttcaaaatat cattaaatca
     2341 caattttttt tttgaatttt ttaatatagt agcctcggat tgactgatta ttgtttgtac
     2401 aaaattttgg tgtctcgttt tctctttgca tatgcaatgt gtcacatgga tgtcttatgt
     2461 ggatttgata ctgatatgtt gtaacgtgat tggtcattta cctaaaaagt gaaatttttc
     2521 tcaaacaatg atctaaattg aaatatagtc ggactttttt tttatgctaa gaatataaat
     2581 atttcatatg taaattagga gaatggatta aatttttagg taacacttaa tgtattaaat
     2641 ccattctcct aatttacata tgttaatatt tacaatatta tcttctattt atcattttta
     2701 atttaatcaa gttattcaaa tctatttatt tttttgtcaa aatttcaaat cttattaata
     2761 taaaaaatta tttgagaaat tatcataaaa caatctaatt aatatgcatt attattggat
     2821 aacgattttt ggtatttaaa acaatctagt tcacttaaat ctatgataga ttgttttaaa
     2881 taccaaaaat cgttatccaa taataatgta tattacaaat ttaaaatagt gcttcaaagg
     2941 aatccaacaa gaaaaataaa aaacaacaca agtacacaac aacaataaca acgcaagtag
     3001 gttaggtact taggtagcaa tgcatgcagc ctgattcaaa cgatctacat atgtttttaa
     3061 ataaaagatc atgtcatttg attcttccca ttgtctattt tctataataa gtaggttagg
     3121 tatcaacgca gcctccaaaa gcataaaaac aaattgcaaa ttttccagtg gcatatgtat
     3181 atattagtca cttgaatttc attaaataaa atacgagtag aaacgttgac aaatattacg
     3241 agtaaacaaa aacaaattgg actttggaag cctttacatt ttgctcacgc aagtaaaaac
     3301 aaaaggaaaa agaaatactc aattatttga catagtagaa ataagatatt catatgaaag
     3361 tgaaagccat ttgatctttc aaaggtttaa tggttttata ttaggctaag gtatgctttt
     3421 caaacaattg tttaactaaa aaaactgtag atacaaaaaa ataatcataa tttcagcata
     3481 ttgcaaataa gattgtttgt tgtgtattac aatttacaaa taaaattata tatcgtaagt
     3541 ttataaccat aaacagaatc tgtctttaat tttttagaag gtcacaactg aggaaagaaa
     3601 ataaaagttt ttttaatgat ttataaaaat tacaaaagtt aagaaatttc aaaataaaaa
     3661 tgtggtaaac aaaacttctt ttatttgtta agacattgct ataatttttt ttcttattaa
     3721 ttatttagtt atactgaaat taaattttat ataattagct gcaaactcat aattacttaa
     3781 cttagtctaa aaaccagtta tggtcataaa ctatatatta tgagaaaatt agtcagtctt
     3841 actttttttt atgaaattat ttcattgcaa gttatattca atctatagat aatgaatatt
     3901 tatttaataa tcatgataag ataagatcaa atatagaaga aaatatacac aagagaaaaa
     3961 aactaaataa aaagtataat taaataagta caagtcatta tccaaaaata tccaccactc
     4021 gcgtaaaagc gcgtggaagt taaaaagtca ttctcctaaa tgaaccaagt cttcaagtct
     4081 caactacttc ctaaagacaa ttagaaattt agaaccaaaa caaacgaaac ttctctgcaa
     4141 tagccttctc caaatttcct cactttcacg cgtttcagtc ttctctatat aaacttgttt
     4201 catcaaccca tcaaaatccc actcaaattc tgtttcagtg ttcttcaaat tctcaatctt
     4261 tctctcaatt catctaaagg gttcctcaaa gtttcaatct ttctctctga atctaaacca
     4321 acatgagacc gaattctcgc cctatagtga gtcgtattac ctggccgtcg ttttacaacg
     4381 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     4441 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     4501 cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4561 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     4621 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     4681 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     4741 accgaaacgc gcga
//
