LOCUS       sequence-396362-        4404 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 4404)
  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 4404)
  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..4404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     386..403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    520..541
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(556..561,562..579,580..586)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    594..610
                     /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        624..642
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     624..641
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2402..2421)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2403..2421)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2431..2453)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2647..2666)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2766..2788
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2826..2844)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2912..3016
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3017..3085,3086..3877)
                     /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(3235..3254)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4048..4404,1..232)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
       61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
      421 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
      481 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
      541 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
      601 cggataacaa tttcacacga gctatttagg tgacactata gaagtgaagc ttggtctcaa
      661 cctttaatcc tcttctcgta catcacattc cttgcactat cttttgtagt tagtgtgata
      721 tttctgtatg gctgtaatgt tttcatgatc atgatcatgt tctttaaatg aacgatgtaa
      781 acggcagctt tgtccaaaac tcacaaagat tacatctaaa agactgccat taccagtctt
      841 tgatgtgtac tgggtcaact ttgaaggccc aaacacatat tctatagtga ggcccaaatc
      901 aagatagata ccgtcttctt ctttttttaa tttcttaaga aaatatgtac cgtacatgtt
      961 ttttgttttt ttgaattgag atttactcct tcaactaatg agtccacacc actgacactt
     1021 cttattttct acgctataag ttttttttat tgagatttct agttttcaaa aattacataa
     1081 gaagaataga cagtgaaact gaaataaaga ctcttacatg ttgcatctat agatttcctt
     1141 ttaaatataa aatatagatt ttaatatgaa aacttgatag ttacaaatct acttaatttt
     1201 aattatatga ttctaactag aagatattga gatttgcaag ataatattct ctatttgtgt
     1261 gtatgaaata aaacagattt ttaattatat ttattcaaaa agtctaatag ctctaataac
     1321 atttgttgaa caatgtctgt ttggtgaaaa agaaattaaa atcaggtgtt gacaacttca
     1381 tgaaaaagaa atgtaatcta aaatctagtc gcttacacat tggcagaata attttctttg
     1441 tgtttacatc atcttcatat atcttagaga tatctcttta taaaaagtta tagtatatat
     1501 attttgtttg gtcaacatac gtaattttat atcaataatt aatgatatag aatgacattt
     1561 taaatgcatt aaatttgaaa tgaaatgaaa aagctagaga agtggagcgc cacaaattga
     1621 aaaggtgata gaaagtgttg gcctctcgag actaattaat aacgtggatt actccaaaac
     1681 ccttaaaagt attacattta ggaaggagaa aagccatttc aagttttcaa tgactgtata
     1741 aatctgatct tttgactgca tgagccttca agtattcaaa tggcaacaga attaaaacga
     1801 gagagaaata gtcacgacgg ccattaatct tgtgaatggt cggttttgat tagtggagaa
     1861 ggagaggtca atgactctct ataatacgat tctctcacca cgtggacaca gcggaattag
     1921 tcccttaatt acatatttta attacagaga gacaacctaa tcacttggcc acgttgaaat
     1981 tgctaggtac tagttaagac ttgttccgca atatctgtct taaaggtaag tgtataaatt
     2041 catttgacta tattttattt tcacggtttt tcttcattct aaaaagtagt ttcatcttct
     2101 tccctcatcc atcatcattc atcatcatat cttctacttc ttcttcttct caggttctct
     2161 tttttgtcct cttttgtgta aatacgtttt gtcatgattc tctgtgaaca gtaccgatcg
     2221 tttttttttg ggattagcga aaaataaaaa taaccgacgt tttttgtatt atttgtctga
     2281 cagacagaat ctgtattatt ttgtcatctt ttttcaatag tctcatgatt gtctcttgtg
     2341 tgtttccagc tttcccatag ggtccattga tttttgcagc caacatgaga ccgaattctc
     2401 gccctatagt gagtcgtatt acctggccgt cgttttacaa cgtcgtgact gggaaaaccc
     2461 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     2521 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg
     2581 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac
     2641 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     2701 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     2761 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     2821 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     2881 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     2941 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3001 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3061 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3121 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3181 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     3241 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     3301 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     3361 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     3421 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     3481 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     3541 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     3601 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     3661 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     3721 cggtgagcgt ggctctcgag gtatcattgc agcactgggg ccagatggta agccctcccg
     3781 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     3841 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     3901 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     3961 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4021 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4081 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4141 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     4201 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4261 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4321 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4381 cacacagccc agcttggagc gaac
//
