LOCUS       sequence-298543-        4791 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0.5 Golden Gate vector, loxP-GUS-nosT-loxP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4791)
  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 4791)
  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: pLOXp-GUS
FEATURES             Location/Qualifiers
     source          1..4791
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(7..40)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(227..244)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(337..925)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(417..436)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1099..1893)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(1894..1998)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2299..2321
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2313..2330
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2314..2330
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2341..2360
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2341..2359
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2364..2375
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(2459..2492)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      4533..4785
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
ORIGIN
        1 gacctaataa cttcgtatag catacattat acgaagttat attaagggtt gaatgtgaga
       61 cgcacagagt ggcggccgct tggatcccag aattctaggt acctcttaat taactggcct
      121 catgggcctt cctttcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
      181 aacatggtca tagctgtttc cttgcgtatt gggcgctctc cgcttcctcg ctcactgact
      241 cgctgcgctc ggtcgttcgg gtaaagcctg gggtgcctaa tgagcaaaag gccagcaaaa
      301 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
      361 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
      421 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
      481 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
      541 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
      601 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
      661 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
      721 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
      781 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
      841 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
      901 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
      961 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     1021 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1081 aacttggtct gacagttatt agaaaaattc atccagcaga cgataaaacg caatacgctg
     1141 gctatccggt gccgcaatgc catacagcac cagaaaacga tccgcccatt cgccgcccag
     1201 ttcttccgca atatcacggg tggccagcgc aatatcctga taacgatccg ccacgcccag
     1261 acggccgcaa tcaataaagc cgctaaaacg gccattttcc accataatgt tcggcaggca
     1321 cgcatcacca tgggtcacca ccagatcttc gccatccggc atgctcgctt tcagacgcgc
     1381 aaacagctct gccggtgcca ggccctgatg ttcttcatcc agatcatcct gatccaccag
     1441 gcccgcttcc atacgggtac gcgcacgttc aatacgatgt ttcgcctgat gatcaaacgg
     1501 acaggtcgcc gggtccaggg tatgcagacg acgcatggca tccgccataa tgctcacttt
     1561 ttctgccggc gccagatggc tagacagcag atcctgaccc ggcacttcgc ccagcagcag
     1621 ccaatcacgg cccgcttcgg tcaccacatc cagcaccgcc gcacacggaa caccggtggt
     1681 ggccagccag ctcagacgcg ccgcttcatc ctgcagctcg ttcagcgcac cgctcagatc
     1741 ggttttcaca aacagcaccg gacgaccctg cgcgctcaga cgaaacaccg ccgcatcaga
     1801 gcagccaatg gtctgctgcg cccaatcata gccaaacaga cgttccaccc acgctgccgg
     1861 gctacccgca tgcaggccat cctgttcaat catactcttc ctttttcaat attattgaag
     1921 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     1981 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat
     2041 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     2101 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtggccgc
     2161 tacagggcgc tcccattcgc cattcaggct gcgcaactgt tgggaagggc gtttcggtgc
     2221 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     2281 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgacg
     2341 taatacgact cactataggg cgaattgaag gaaggccgtc aaggcctagg cgcgccatga
     2401 gctctaaagc ttcgctcgag ctgcggccgc cacttcgtgc acgtctcata ctgacctaat
     2461 aacttcgtat agcatacatt atacgaagtt atattaaggg ttgaatgggt cagtccctta
     2521 tgttacgtcc tgtagaaacc ccaacccgtg aaatcaaaaa actcgacggc ctgtgggcat
     2581 tcagtctgga tcgcgaaaac tgtggaattg atcagcgttg gtgggaaagc gcgttacaag
     2641 aaagccgggc aattgctgtg ccaggcagtt ttaacgatca gttcgccgat gcagatattc
     2701 gtaattatgc gggcaacgtc tggtatcagc gcgaagtctt tataccgaaa ggtaagtctt
     2761 actctctctt ttttggtctg tatttttaat tttttgaagt atactatttg tactgacgct
     2821 aataatcttt tttcaggttg ggcaggccag cgtatcgtgc tgcgtttcga tgcggtcact
     2881 cattacggca aagtgtgggt caataatcag gaagtgatgg agcatcaggg cggctatacg
     2941 ccatttgaag ccgatgtcac gccgtatgtt attgccggga aaagtgtacg tatcaccgtt
     3001 tgtgtgaaca acgaactgaa ctggcagact atcccgccgg gaatggtgat taccgacgaa
     3061 aacggcaaga aaaagcagtc ttacttccat gatttcttta actatgccgg aatccatcgc
     3121 agcgtaatgc tctacaccac gccgaacacc tgggtggacg atatcaccgt ggtgacgcat
     3181 gtcgcgcaag actgtaacca cgcgtctgtt gactggcagg tacttcatgc ttcaacgtgt
     3241 aacttaagag atactgtgtg aaattttata tttccataca tttgcttgac ctttgctttt
     3301 tgtcaatttt tttcccctta caggtggtgg ccaatggtga tgtcagcgtt gaactgcgtg
     3361 atgcggatca acaggtggtt gcaactggac aaggcactag cgggactttg caagtggtga
     3421 atccgcacct ctggcaaccg ggtgaaggtt atctctatga actgtgcgtc acagccaaaa
     3481 gccagacaga gtgtgatatc tacccgcttc gcgtcggcat ccggtcagtg gcagtgaagg
     3541 gcgaacagtt cctgattaac cacaaaccgt tctactttac tggctttggt cgtcatgaag
     3601 atgcggactt gcgtggcaaa ggattcgata acgtgctgat ggtgcacgac cacgcattaa
     3661 tggactggat tggggccaac tcctaccgta cctcgcatta cccttacgct gaagagatgc
     3721 tcgactgggc agatgaacat ggcatcgtgg tgattgatga aactgctgct gtcggcttta
     3781 acctctcttt aggcattggt ttcgaagcgg gcaacaagcc gaaagaactg tacagcgaag
     3841 aggcagtcaa cggggaaact cagcaagcgc acttacaggc gattaaagag ctgatagcgc
     3901 gtgacaaaaa ccacccaagc gtggtgatgt ggagtattgc caacgaaccg gatacccgtc
     3961 cgcaaggtgc acgggaatat ttcgcgccac tggcggaagc aacgcgtaaa ctcgacccga
     4021 cgcgtccgat cacctgcgtc aatgtaatgt tctgcgacgc tcacaccgat accatcagcg
     4081 atctctttga tgtgctgtgc ctgaaccgtt attacggatg gtatgtccaa agcggcgatt
     4141 tggaaacggc agagaaggta ctggaaaaag aacttctggc ctggcaggag aaactgcatc
     4201 agccgattat catcaccgaa tacggcgtgg atacgttagc cgggctgcac tcaatgtaca
     4261 ccgacatgtg gagtgaagag tatcagtgtg catggctgga tatgtatcac cgcgtctttg
     4321 atcgcgtcag cgccgtcgtc ggtgaacagg tatggaattt cgccgatttt gcgacctcgc
     4381 aaggcatatt gcgcgttggc ggtaacaaga aagggatctt cactcgcgac cgcaaaccga
     4441 agtcggcggc ttttctgctg caaaaacgct ggactggcat gaacttcggt gaaaaaccgc
     4501 agcagggagg caaacaatga gcttgtcaag cagatcgttc aaacatttgg caataaagtt
     4561 tcttaagatt gaatcctgtt gccggtcttg cgatgattat catataattt ctgttgaatt
     4621 acgttaagca tgtaataatt aacatgtaat gcatgacgtt atttatgaga tgggttttta
     4681 tgattagagt cccgcaatta tacatttaat acgcgataga aaacaaaata tagcgcgcaa
     4741 actaggataa attatcgcgc gcggtgtcat ctatgttact agatcgacgc t
//
