LOCUS       sequence-453812-        3084 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cloning vector to clone promoter fragment in front of minimal 
            p35S:Luciferase:tNOS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3084)
  AUTHORS   Winkelmolen T, Colleoni P, Moscou M, Hoseinzadeh P, Oldach K, 
            Schmidt RC, Immink R, van Esse GW
  TITLE     VRS5 (HvTB1) binds to the promoter of tillering and floral homeotic 
            genes to regulate their expression
  JOURNAL   bioRxiv 2024.08.22.609225
REFERENCE   2  (bases 1 to 3084)
  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: "bioRxiv 
            2024.08.22.609225"
COMMENT     Sequence Label: L1p1_min35S_Luciferase
FEATURES             Location/Qualifiers
     source          1..3084
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    114..135
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(150..155,156..173,174..180)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    188..204
                     /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)."
     RBS             222..233
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             240..917
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      962..1033
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1049..1076
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        1094..1140
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             1141..2793
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(1207..1225)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     terminator      2794..3046
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    complement(3051..3075)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 ggagtacgag acgcaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
       61 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
      121 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
      181 tgtggaattg tgagcggata acaatttcac acatactaga gaaagaggag aaatactaga
      241 tggcttcctc cgaggacgtt atcaaagagt tcatgcgttt caaagttcgt atggaaggtt
      301 ccgttaacgg tcacgagttc gaaatcgaag gtgaaggtga aggtcgtccg tacgaaggta
      361 cccagaccgc taaactgaaa gttaccaaag gtggtccgct gccgttcgct tgggacatcc
      421 tgtccccgca gttccagtac ggttccaaag cttacgttaa acacccggct gacatcccgg
      481 actacctgaa actgtccttc ccggaaggtt tcaaatggga acgtgttatg aacttcgagg
      541 acggtggtgt tgttaccgtt acccaggact cctccctgca agacggtgag ttcatctaca
      601 aagttaaact gcgtggtacc aacttcccgt ccgacggtcc ggttatgcag aaaaaaacca
      661 tgggttggga agcttccacc gaacgtatgt acccggagga cggtgctctg aaaggtgaaa
      721 tcaaaatgcg tctgaaactg aaagacggtg gtcactacga cgctgaagtt aaaaccacct
      781 acatggctaa aaaaccggtt cagctgccgg gtgcttacaa aaccgacatc aaactggaca
      841 tcacctccca caacgaggac tacaccatcg ttgaacagta cgaacgtgct gaaggtcgtc
      901 actccaccgg tgcttaataa cgctgatagt gctagtgtag atcgctacta gagccaggca
      961 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     1021 ggtgaacgct ctctactaga gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta
     1081 tacgtctccg caagcaagac ccttcctcta tataaggaag ttcatttcat ttggagagga
     1141 atggaggacg ccaaaaacat aaagaaaggc ccggcgccat tctatcctct agaggatgga
     1201 accgctggag agcaactgca taaggctatg aagagatacg ccctggttcc tggaacaatt
     1261 gcttttacag atgcacatat cgaggtgaac atcacgtacg cggaatactt cgaaatgtcc
     1321 gttcggttgg cagaagctat gaaacgatat gggctgaata caaatcacag aatcgtcgta
     1381 tgcagtgaaa actctcttca attctttatg ccggtgttgg gcgcgttatt tatcggagtt
     1441 gcagttgcgc ccgcgaacga catttataat gaacgtgaat tgctcaacag tatgaacatt
     1501 tcgcagccta ccgtagtgtt tgtttccaaa aaggggttgc aaaaaatttt gaacgtgcaa
     1561 aaaaaattac caataatcca gaaaattatt atcatggatt ctaaaacgga ttaccaggga
     1621 tttcagtcga tgtacacgtt cgtcacatct catctacctc ccggttttaa tgaatacgat
     1681 tttgtaccag agtcctttga tcgtgacaaa acaattgcac tgataatgaa ttcctctgga
     1741 tctactgggt tacctaaggg tgtggccctt ccgcatagaa ctgcctgcgt cagattctcg
     1801 catgccagag atcctatttt tggcaatcaa atcattccgg atactgcgat tttaagtgtt
     1861 gttccattcc atcacggttt tggaatgttt actacactcg gatatttgat atgtggattt
     1921 cgagtcgtct taatgtatag atttgaagaa gagctgtttt tacgatccct tcaggattac
     1981 aaaattcaaa gtgcgttgct agtaccaacc ctattttcat tcttcgccaa aagcactctg
     2041 attgacaaat acgatttatc taatttacac gaaattgctt ctgggggcgc acctctttcg
     2101 aaagaagtcg gggaagcggt tgcaaaacgc ttccatcttc cagggatacg acaaggatat
     2161 gggctcactg agactacatc agctattctg attacacccg agggggatga taaaccgggc
     2221 gcggtcggta aagttgttcc attttttgaa gcgaaggttg tggatctgga taccgggaaa
     2281 acgctgggcg ttaatcagag aggcgaatta tgtgtcagag gacctatgat tatgtccggt
     2341 tatgtaaaca atccggaagc gaccaacgcc ttgattgaca aggatggatg gctacattct
     2401 ggagacatag cttactggga cgaggacgaa cacttcttca tagttgaccg cttgaagtct
     2461 ttaattaaat acaaaggata tcaggtggcc cccgctgaat tggaatcgat attgttacaa
     2521 caccccaaca tcttcgacgc gggcgtggca ggtctgcccg acgatgacgc cggtgaactt
     2581 cccgccgccg ttgttgtttt ggagcacgga aagacgatga cggaaaaaga gatcgtggat
     2641 tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt gtttgtggac
     2701 gaagtaccga aaggtcttac cggaaaactc gacgcaagaa aaatcagaga gatcctcata
     2761 aaggccaaga agggcggaaa gtccaaattg taagatcgtt caaacatttg gcaataaagt
     2821 ttcttaagat tgaatcctgt tgccggtctt gcgatgatta tcatataatt tctgttgaat
     2881 tacgttaagc atgtaataat taacatgtaa tgcatgacgt tatttatgag atgggttttt
     2941 atgattagag tcccgcaatt atacatttaa tacgcgatag aaaacaaaat atagcgcgca
     3001 aactaggata aattatcgcg cgcggtgtca tctatgttac tagatcctag acccagcttt
     3061 cttgtacaaa gttggcatta cgct
//
