LOCUS       sequence-388372-        6035 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains NET4A promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6035)
  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 6035)
  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-PIP74
FEATURES             Location/Qualifiers
     source          1..6035
                     /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(5621..5640)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5622..5640)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5650..5672)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5866..5885)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5985..6007
                     /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 tatttagagg gtgttattgg tttgtgattt
     2281 atatagattt taaatgattt gtaatgtcat gaattttttt ttgttattta attaaaattt
     2341 tgtaagattt ttttaaagtt tggtgttatt agattttgat ttgtaaaaaa gaaatttaaa
     2401 gtttttgtaa atctggagtt attagattga tactttaata aaatcattac aaattttgtg
     2461 ttattcaact taaacaacaa tttccatgac tgttaatgat tataaaatat tgtgttattc
     2521 gattatgatt ttatcttatt ttcttcacaa attaaaatca tgaaaagtat cataaaaaaa
     2581 gagagattgt gaatcatgat tttggatata aacaaaaacc aatgatacga acagatgcat
     2641 aaatctcatt aacagtgatg aacaatatat atattctaat gggtcaagtc ccacgaatgc
     2701 tcctctataa tctctttgtg aaaaatcaac acacaagcaa tttttaaaac aaagcatttt
     2761 aatctaacat atgaaaagta aatatccaac aaaatatttg aaaataaaac atcaagttct
     2821 tcatggatgg ttccctattc atgtaacttt acttacaaat accatagtta ttgtgtttat
     2881 gcgtgtgatt ttcattttga tgaaaataac taagaatagt atactatatt ttggagatat
     2941 gttataattt tgaaaaaatc acaaattaaa gtttataatt atattcaagg agattttaca
     3001 agattataga aagactttac aaaattttag aaaaacaaat cataaaaatt atacaaaact
     3061 ctctagtaat catttaaaat cttttaattt atctattttt tatatatttt aaaagtcatt
     3121 aaaacttttt cacaatcaca aaccaataaa ttaatgaatt acaattttga tcatagcaaa
     3181 agaaaatgat agacttcact acttcagcta attgattacg ttgttgctct tgttgaattc
     3241 tcaagagctg cattgaccaa tcacttcgtt tgctttgaac aataatcttc aaagagagat
     3301 gtgtttctat ttcatagagt ttcatcatca aagagcactc aactaatctc ttttggacat
     3361 cgctcatcct cggttaacca taacaaaaag ttatgaatat tcacagacat agaaatacca
     3421 gaaaattcgt acagatcaag tgagaggatt tatttgaaag agtcaagact tattgcatag
     3481 aaacaaatta taatattaat cacgtaaata atactactac ttacaaaaag agtaaacttc
     3541 acttgatata tacttatact acaagatatt caatttttgt atttacaaga acttgtggga
     3601 aaaaaataaa ttaagaagat attataacga cattaactct gacaaacttt gtaggtaggg
     3661 attggaattt agcaaaaaac tttgtaggta tgagaaacta ctggagcaag aaactacgag
     3721 tagagaaagt tagaatttaa gtcaggtgag ctgttggcat gtcacatggg acgcaatgtg
     3781 gatttcatgt gcattgttct ttttatatat ctctacatct cattctgtcg ctatccgatt
     3841 tccgatccac aagctctaca agaccaacac attcttgttt ttgtaccctt ctaatttctc
     3901 atatttacag ctatatgctg ttcttagcgt aatttattta tcacgtcttg tttatgatct
     3961 aaatgatggt gatataaggg cgacattggt tttgttattg atatcattag tagtgctttt
     4021 ggtttcgggt tttcttttgc attgttatag aaagtagaaa cgtaagcatg attaatgttc
     4081 catggaaaat ttagtactaa gcgaaaccat cgaaacatca cttttttatt gttatacaag
     4141 caacgacaaa gttgagaaaa caatatttaa gttatccgat tattagttta gattttccga
     4201 atagcaaact gatatattta ttgattgaca gtagatcgac agatgtaaga acttatagaa
     4261 tattttaaag atttcaaagt agcaatttag catcagtagt taaatctaaa cgagagtatc
     4321 caatgcgtgt gttttacatt atcgtcctaa ctacggtaat atgcatcaac ttttttcttc
     4381 aactctctcg tatagatctg atggaccaaa aaacaatttc tgcatatatt tcatataaac
     4441 tgattgctat gacatgcatg caaaagaaag atttaatata tattttacga aagtaactaa
     4501 acttaactaa ctagttaata taggtacgaa ataaaccatt aagaattcat ctactctccg
     4561 accatggtac ggtgatttac tattctccgc attgtcatac gcaatgagac catttttttt
     4621 tcccgtggtt cttagctcga ttttatactg tcacacactc attacaaggt attgctcctg
     4681 cctgttccta cggcttttga tgagtttgtt tatgcatttg acgtgtatta agtgtgtaac
     4741 caaagagagc ttctctaaaa ttgtaagttg cgtttggaaa ataataagaa tcttgacatt
     4801 actaccgaaa tacttatctt ttctatgtta gaccaatggt ccaagactcc aatgttagag
     4861 atttttttac tattcatgtg acaacaataa ttggtatgta ggagtgaagg aagaaccgac
     4921 aatcacaatc caattaacat aaagtagctc ccacatctta cccaacgcca tcctcaacgt
     4981 tgatatctca ccatccaacg gcatccagct tccgatagat taaaccaatc acagccgttg
     5041 gatcaaattt tacctttccc tcttcttact ctttcataaa taaatccaac ttcttcactc
     5101 tccacccgtc tcacttcctc tcctactccg gtcgacgatt atcccaatct cgccgggatt
     5161 ctcctctgca ccgtgagtta ttgtttactc tgctatttcc tttttatgta ttgtttcctc
     5221 tctagggaat gagattatga actcctacta cttattgtga tactttgatc gaatcgtctt
     5281 cctccaaatc tgtatccgct cttgatctgt ttcatatttt ctctcttaac ttggattttg
     5341 attttctatc acttgaaaat gtgttgccgt tttgaattcc gtgaagaatt tgaaatcgga
     5401 gttttcttct ctgcggcgtt gtgttttttt ccgattgatt tgttttggga tctgatacag
     5461 attgtttgat ttcagatctt cgagatttag ttgttgacat tcgatataat ttcatttgga
     5521 tctgatacag attcttttgc tttcaaaatc cgttgtttac tgttttgttc atgtaatgtt
     5581 gatttcagat aaaatagaag aacatgagac cgaattctcg ccctatagtg agtcgtatta
     5641 cctggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     5701 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     5761 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct
     5821 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
     5881 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     5941 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     6001 tcagaggttt tcaccgtcat caccgaaacg cgcga
//
