LOCUS       sequence-312899-        4710 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 4710)
  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 4710)
  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..4710
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(366..470)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     771..793
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     785..802
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     786..802
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     813..832
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        813..831
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             836..847
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(891..924)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      2965..3217
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    complement(3230..3263)
                     /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(3409..3426)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3519..4107)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3599..3618)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4281..4710,1..365))
                     /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"
ORIGIN
        1 cgacgcatgg catccgccat aatgctcact ttttctgccg gcgccagatg gctagacagc
       61 agatcctgac ccggcacttc gcccagcagc agccaatcac ggcccgcttc ggtcaccaca
      121 tccagcaccg ccgcacacgg aacaccggtg gtggccagcc agctcagacg cgccgcttca
      181 tcctgcagct cgttcagcgc accgctcaga tcggttttca caaacagcac cggacgaccc
      241 tgcgcgctca gacgaaacac cgccgcatca gagcagccaa tggtctgctg cgcccaatca
      301 tagccaaaca gacgttccac ccacgctgcc gggctacccg catgcaggcc atcctgttca
      361 atcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
      421 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
      481 cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
      541 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
      601 aagaatagac cgagataggg ttgagtggcc gctacagggc gctcccattc gccattcagg
      661 ctgcgcaact gttgggaagg gcgtttcggt gcgggcctct tcgctattac gccagctggc
      721 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
      781 acgttgtaaa acgacggcca gtgagcgcga cgtaatacga ctcactatag ggcgaattga
      841 aggaaggccg tcaaggccgc atcacttcgt gcacgtctca tactgaccta ataacttcgt
      901 atagcataca ttatacgaag ttatattaag ggttgaatgg gtcagtccct tatgttacgt
      961 cctgtagaaa ccccaacccg tgaaatcaaa aaactcgacg gcctgtgggc attcagtctg
     1021 gatcgcgaaa actgtggaat tgatcagcgt tggtgggaaa gcgcgttaca agaaagccgg
     1081 gcaattgctg tgccaggcag ttttaacgat cagttcgccg atgcagatat tcgtaattat
     1141 gcgggcaacg tctggtatca gcgcgaagtc tttataccga aaggtaagtc ttactctctc
     1201 ttttttggtc tgtattttta attttttgaa gtatactatt tgtactgacg ctaataatct
     1261 tttttcaggt tgggcaggcc agcgtatcgt gctgcgtttc gatgcggtca ctcattacgg
     1321 caaagtgtgg gtcaataatc aggaagtgat ggagcatcag ggcggctata cgccatttga
     1381 agccgatgtc acgccgtatg ttattgccgg gaaaagtgta cgtatcaccg tttgtgtgaa
     1441 caacgaactg aactggcaga ctatcccgcc gggaatggtg attaccgacg aaaacggcaa
     1501 gaaaaagcag tcttacttcc atgatttctt taactatgcc ggaatccatc gcagcgtaat
     1561 gctctacacc acgccgaaca cctgggtgga cgatatcacc gtggtgacgc atgtcgcgca
     1621 agactgtaac cacgcgtctg ttgactggca ggtacttcat gcttcaacgt gtaacttaag
     1681 agatactgtg tgaaatttta tatttccata catttgcttg acctttgctt tttgtcaatt
     1741 tttttcccct tacaggtggt ggccaatggt gatgtcagcg ttgaactgcg tgatgcggat
     1801 caacaggtgg ttgcaactgg acaaggcact agcgggactt tgcaagtggt gaatccgcac
     1861 ctctggcaac cgggtgaagg ttatctctat gaactgtgcg tcacagccaa aagccagaca
     1921 gagtgtgata tctacccgct tcgcgtcggc atccggtcag tggcagtgaa gggcgaacag
     1981 ttcctgatta accacaaacc gttctacttt actggctttg gtcgtcatga agatgcggac
     2041 ttgcgtggca aaggattcga taacgtgctg atggtgcacg accacgcatt aatggactgg
     2101 attggggcca actcctaccg tacctcgcat tacccttacg ctgaagagat gctcgactgg
     2161 gcagatgaac atggcatcgt ggtgattgat gaaactgctg ctgtcggctt taacctctct
     2221 ttaggcattg gtttcgaagc gggcaacaag ccgaaagaac tgtacagcga agaggcagtc
     2281 aacggggaaa ctcagcaagc gcacttacag gcgattaaag agctgatagc gcgtgacaaa
     2341 aaccacccaa gcgtggtgat gtggagtatt gccaacgaac cggatacccg tccgcaaggt
     2401 gcacgggaat atttcgcgcc actggcggaa gcaacgcgta aactcgaccc gacgcgtccg
     2461 atcacctgcg tcaatgtaat gttctgcgac gctcacaccg ataccatcag cgatctcttt
     2521 gatgtgctgt gcctgaaccg ttattacgga tggtatgtcc aaagcggcga tttggaaacg
     2581 gcagagaagg tactggaaaa agaacttctg gcctggcagg agaaactgca tcagccgatt
     2641 atcatcaccg aatacggcgt ggatacgtta gccgggctgc actcaatgta caccgacatg
     2701 tggagtgaag agtatcagtg tgcatggctg gatatgtatc accgcgtctt tgatcgcgtc
     2761 agcgccgtcg tcggtgaaca ggtatggaat ttcgccgatt ttgcgacctc gcaaggcata
     2821 ttgcgcgttg gcggtaacaa gaaagggatc ttcactcgcg accgcaaacc gaagtcggcg
     2881 gcttttctgc tgcaaaaacg ctggactggc atgaacttcg gtgaaaaacc gcagcaggga
     2941 ggcaaacaat gagcttgtca agcagatcgt tcaaacattt ggcaataaag tttcttaaga
     3001 ttgaatcctg ttgccggtct tgcgatgatt atcatataat ttctgttgaa ttacgttaag
     3061 catgtaataa ttaacatgta atgcatgacg ttatttatga gatgggtttt tatgattaga
     3121 gtcccgcaat tatacattta atacgcgata gaaaacaaaa tatagcgcgc aaactaggat
     3181 aaattatcgc gcgcggtgtc atctatgtta ctagatcgac gctgacctaa taacttcgta
     3241 tagcatacat tatacgaagt tatattaagg gttgaatgtg agacgcacag agtgctgggc
     3301 ctcatgggcc ttcctttcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     3361 ttaacatggt catagctgtt tccttgcgta ttgggcgctc tccgcttcct cgctcactga
     3421 ctcgctgcgc tcggtcgttc gggtaaagcc tggggtgcct aatgagcaaa aggccagcaa
     3481 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3541 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3601 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3661 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3721 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3781 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3841 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3901 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3961 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4021 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4081 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4141 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4201 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4261 taaacttggt ctgacagtta ttagaaaaat tcatccagca gacgataaaa cgcaatacgc
     4321 tggctatccg gtgccgcaat gccatacagc accagaaaac gatccgccca ttcgccgccc
     4381 agttcttccg caatatcacg ggtggccagc gcaatatcct gataacgatc cgccacgccc
     4441 agacggccgc aatcaataaa gccgctaaaa cggccatttt ccaccataat gttcggcagg
     4501 cacgcatcac catgggtcac caccagatct tcgccatccg gcatgctcgc tttcagacgc
     4561 gcaaacagct ctgccggtgc caggccctga tgttcttcat ccagatcatc ctgatccacc
     4621 aggcccgctt ccatacgggt acgcgcacgt tcaatacgat gtttcgcctg atgatcaaac
     4681 ggacaggtcg ccgggtccag ggtatgcaga
//
