LOCUS       sequence-313097-        3471 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 Golden Gate vector, CRE with U5 intron at 254bp, in 
            pICH41308 backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3471)
  AUTHORS   Harrington SA, Backhaus AE, Fox S, Rogers C, Borrill P, Uauy C, 
            Richardson A
  TITLE     A heat-shock inducible system for flexible gene expression in 
            cereals.
  JOURNAL   Plant Methods. 2020 Oct 14;16:137. doi: 10.1186/s13007-020-00677-3. 
            eCollection 2020.
  PUBMED    33072173
REFERENCE   2  (bases 1 to 3471)
  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: "Plant 
            Methods."; date: "2020-10-14"; volume: "16:137. doi"; pages: " 
            10.1186/s13007-020-00677-3. eCollection 2020"
COMMENT     Sequence Label: EC71171
FEATURES             Location/Qualifiers
     source          1..3471
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          79..267
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     complement(1102..1120)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1188..1292
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1514..2305
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      2476..3064
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2965..2984
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3218..3235
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcatggtgca agttgaataa ccggaaatgg tttcccgcag aacctgaaga tgttcgcgat
       61 tatcttctat atcttcaggt aagtttctgc ttctaccttt gatatatata taataattat
      121 cattaattag tagtaatata atatttcaaa tatttttttc aaaataaaag aatgtggtat
      181 atagcaattg cttttctgta gtttataagt gtgtatattt taatttataa cttttctaat
      241 atatgaccaa aatttgttga tgtgcaggcg cgcggtctgg cagtaaaaac tatccagcaa
      301 catttgggcc agctaaacat gcttcatcgt cggtccgggc tgccacgacc aagtgacagc
      361 aatgctgttt cactggttat gcggcggatc cgaaaagaaa acgttgatgc cggtgaacgt
      421 gcaaaacagg ctctagcgtt cgaacgcact gatttcgacc aggttcgttc actcatggaa
      481 aatagcgatc gctgccagga tatacgtaat ctggcatttc tggggattgc ttataacacc
      541 ctgttacgta tagccgaaat tgccaggatc agggttaaag atatctcacg tactgacggt
      601 gggagaatgt taatccatat tggcagaacg aaaacgctgg ttagcaccgc aggtgtagag
      661 aaggcactta gcctgggggt aactaaactg gtcgagcgat ggatttccgt ttctggtgta
      721 gctgatgatc cgaataacta cctgttttgc cgggtcagaa aaaatggtgt tgccgcgcca
      781 tctgccacca gccagctatc aactcgcgcc ctggaaggga tttttgaagc aactcatcga
      841 ttgatttacg gcgctaagga tgactctggt cagagatacc tggcctggtc tggacacagt
      901 gcccgtgtcg gagccgcgcg agatatggcc cgcgctggag tttcaatacc ggagatcatg
      961 caagctggtg gctggaccaa tgtaaatatt gtcatgaact atatccgtaa cctggatagt
     1021 gaaacagggg caatggtgcg cctgctggaa gatggcgatt aggctttgag accacgaagt
     1081 ggctcttcag tggacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     1141 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     1201 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     1261 aaatgcttca ataatattga aaaaggaaga gtatgcgctc acgcaactgg tccagaacct
     1321 tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt
     1381 tttttttggg gtacagtcta tgcctcgggc atccaagcag caagcgcgtt acgccgtggg
     1441 tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagggcagtc gccctaaaac
     1501 aaagttaaac atcatgaggg aagcggtgat cgccgaagta tcgactcaac tatcagaggt
     1561 agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt tgtacggctc
     1621 cgcagtggat ggcggcctga agccacacag cgatattgat ttgctggtta cggtgaccgt
     1681 aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa cttcggcttc
     1741 ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc acgacgacat
     1801 cattccgtgg cgttatccag ctaagcgcga actgcaattt ggagaatggc agcgcaatga
     1861 cattcttgca ggtatcttcg agccagccac gatcgacatt gatctggcta tcttgctgac
     1921 aaaagcaaga gaacatagcg ttgccttggt aggtccagcg gcggaggaac tctttgatcc
     1981 ggttcctgaa caggatctat ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc
     2041 gcccgactgg gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca tttggtacag
     2101 cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct
     2161 gccggcccag tatcagcccg tcatacttga agctagacag gcttatcttg gacaagaaga
     2221 agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gtccattacg taaaaggcga
     2281 gatcaccaag gtagtcggca aataactgtc agaccaagtt tactcatata tactttagat
     2341 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     2401 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     2461 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2521 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2581 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     2641 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2701 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2761 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2821 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2881 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2941 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3001 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3061 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3121 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     3181 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     3241 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatcact
     3301 ctgtggtctc aaatgtccaa tttactgacc gtacaccaaa atttgcctgc attaccggtc
     3361 gatgcaacga gtgatgaggt tcgcaagaac ctgatggaca tgttcaggga tcgccaggcg
     3421 ttttctgagc atacctggaa aatgctcctg tccgtttgcc ggtcgtgggc g
//
