LOCUS       sequence-464288-        7448 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning vector to clone promoter fragment in front of minimal 
            p35S:Luciferase:tNOS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7448)
  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 7448)
  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..7448
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    76..100
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      192..823
                     /label=oriV
                     /note="incP origin of replication"
     promoter        910..1014
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1015..1083,1084..1875)
                     /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(1233..1252)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2046..2634
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2535..2554
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2970..4118
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     misc_feature    4274..4298
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     protein_bind    4349..4373
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      complement(4378..4630)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(4631..6283)
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     6199..6217
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     promoter        complement(6284..6330)
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     terminator      complement(6344..6371)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(6387..6458)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(6503..7180)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             7187..7198
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    7216..7232
                     /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        complement(join(7240..7246,7247..7264,7265..7270))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7285..7306
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gtggggccca ctgcatccac cccagtacat taaaaacgtc cgcaatgtgt tattaagttg
       61 tctaagcgtc aatttgttta caccacaata tatcctgcca ccagccagcc aacagctccc
      121 cgaccggcag ctcggcacaa aatcaccact cgatacaggc agcccatcag tcagatcagg
      181 atctcctttg cgacgctcac cgggctggtt gccctcgccg ctgggctggc ggccgtctat
      241 ggccctgcaa acgcgccaga aacgccgtcg aagccgtgtg cgagacaccg cggccgccgg
      301 cgttgtggat acctcgcgga aaacttggcc ctcactgaca gatgaggggc ggacgttgac
      361 acttgagggg ccgactcacc cggcgcggcg ttgacagatg aggggcaggc tcgatttcgg
      421 ccggcgacgt ggagctggcc agcctcgcaa atcggcgaaa acgcctgatt ttacgcgagt
      481 ttcccacaga tgatgtggac aagcctgggg ataagtgccc tgcggtattg acacttgagg
      541 ggcgcgacta ctgacagatg aggggcgcga tccttgacac ttgaggggca gagtgctgac
      601 agatgagggg cgcacctatt gacatttgag gggctgtcca caggcagaaa atccagcatt
      661 tgcaagggtt tccgcccgtt tttcggccac cgctaacctg tcttttaacc tgcttttaaa
      721 ccaatattta taaaccttgt ttttaaccag ggctgcgccc tgtgcgcgtg accgcgcacg
      781 ccgaaggggg gtgccccccc ttctcgaacc ctcccggccc gctaacgcgg gcctcccatc
      841 cccccagggg ctgcgcccct cggccgcgaa cggcctcacc ccaaaaatgg cagcgctggc
      901 caattcgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      961 ctcatgagac aataaccttg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     1021 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     1081 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     1141 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     1201 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     1261 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1321 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1381 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1441 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1501 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1561 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1621 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1681 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg ttctcgcggt
     1741 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1801 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1861 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1921 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1981 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2041 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2101 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2161 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     2221 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2281 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2341 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2401 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2461 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2521 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2581 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2641 agcaacgcgg cctttttacg gttcctggca gatcctagat gtggcgcaac gatgccggcg
     2701 acaagcagga gcgcaccgac ttcttccgca tcaagtgttt tggctctcag gccgaggccc
     2761 acggcaagta tttgggcaag gggtcgctgg tattcgtgca gggcaagatt cggaatacca
     2821 agtacgagaa ggacggccag acggtctacg ggaccgactt cattgccgat aaggtggatt
     2881 atctggacac caaggcacca ggcgggtcaa atcaggaata agggcacatt gccccggcgt
     2941 gagtcggggc aatcccgcaa ggagggtgaa tgaatcggac gtttgaccgg aaggcataca
     3001 ggcaagaact gatcgacgcg gggttttccg ccgaggatgc cgaaaccatc gcaagccgca
     3061 ccgtcatgcg tgcgccccgc gaaaccttcc agtccgtcgg ctcgatggtc cagcaagcta
     3121 cggccaagat cgagcgcgac agcgtgcaac tggctccccc tgccctgccc gcgccatcgg
     3181 ccgccgtgga gcgttcgcgt cgtcttgaac aggaggcggc aggtttggcg aagtcgatga
     3241 ccatcgacac gcgaggaact atgacgacca agaagcgaaa aaccgccggc gaggacctgg
     3301 caaaacaggt cagcgaggcc aagcaggccg cgttgctgaa acacacgaag cagcagatca
     3361 aggaaatgca gctttccttg ttcgatattg cgccgtggcc ggacacgatg cgagcgatgc
     3421 caaacgacac ggcccgctct gccctgttca ccacgcgcaa caagaaaatc ccgcgcgagg
     3481 cgctgcaaaa caaggtcatt ttccacgtca acaaggacgt gaagatcacc tacaccggcg
     3541 tcgagctgcg ggccgacgat gacgaactgg tgtggcagca ggtgttggag tacgcgaagc
     3601 gcacccctat cggcgagccg atcaccttca cgttctacga gctttgccag gacctgggct
     3661 ggtcgatcaa tggccggtat tacacgaagg ccgaggaatg cctgtcgcgc ctacaggcga
     3721 cggcgatggg cttcacgtcc gaccgcgttg ggcacctgga atcggtgtcg ctgctgcacc
     3781 gcttccgcgt cctggaccgt ggcaagaaaa cgtcccgttg ccaggtcctg atcgacgagg
     3841 aaatcgtcgt gctgtttgct ggcgaccact acacgaaatt catatgggag aagtaccgca
     3901 agctgtcgcc gacggcccga cggatgttcg actatttcag ctcgcaccgg gagccgtacc
     3961 cgctcaagct ggaaaccttc cgcctcatgt gcggatcgga ttccacccgc gtgaagaagt
     4021 ggcgcgagca ggtcggcgaa gcctgcgaag agttgcgagg cagcggcctg gtggaacacg
     4081 cctgggtcaa tgatgacctg gtgcattgca aacgctaggg ccttgtgggg tcagttccgg
     4141 ctgggggttc agcagccagc gcctgatctg gggaaccctg tggttggcat gcacatacaa
     4201 atggacgaac ggataaacct tttcacgccc ttttaaatat ccgattattc taataaacgc
     4261 tcttttctct taggtttacc cgccaatata tcctgtcaaa cactgatagt ttaaaccact
     4321 tcgtgcagaa gacaattgca gcgtaatgcc aactttgtac aagaaagctg ggtctaggat
     4381 ctagtaacat agatgacacc gcgcgcgata atttatccta gtttgcgcgc tatattttgt
     4441 tttctatcgc gtattaaatg tataattgcg ggactctaat cataaaaacc catctcataa
     4501 ataacgtcat gcattacatg ttaattatta catgcttaac gtaattcaac agaaattata
     4561 tgataatcat cgcaagaccg gcaacaggat tcaatcttaa gaaactttat tgccaaatgt
     4621 ttgaacgatc ttacaatttg gactttccgc ccttcttggc ctttatgagg atctctctga
     4681 tttttcttgc gtcgagtttt ccggtaagac ctttcggtac ttcgtccaca aacacaactc
     4741 ctccgcgcaa ctttttcgcg gttgttactt gactggcgac gtaatccacg atctcttttt
     4801 ccgtcatcgt ctttccgtgc tccaaaacaa caacggcggc gggaagttca ccggcgtcat
     4861 cgtcgggcag acctgccacg cccgcgtcga agatgttggg gtgttgtaac aatatcgatt
     4921 ccaattcagc gggggccacc tgatatcctt tgtatttaat taaagacttc aagcggtcaa
     4981 ctatgaagaa gtgttcgtcc tcgtcccagt aagctatgtc tccagaatgt agccatccat
     5041 ccttgtcaat caaggcgttg gtcgcttccg gattgtttac ataaccggac ataatcatag
     5101 gtcctctgac acataattcg cctctctgat taacgcccag cgttttcccg gtatccagat
     5161 ccacaacctt cgcttcaaaa aatggaacaa ctttaccgac cgcgcccggt ttatcatccc
     5221 cctcgggtgt aatcagaata gctgatgtag tctcagtgag cccatatcct tgtcgtatcc
     5281 ctggaagatg gaagcgtttt gcaaccgctt ccccgacttc tttcgaaaga ggtgcgcccc
     5341 cagaagcaat ttcgtgtaaa ttagataaat cgtatttgtc aatcagagtg cttttggcga
     5401 agaatgaaaa tagggttggt actagcaacg cactttgaat tttgtaatcc tgaagggatc
     5461 gtaaaaacag ctcttcttca aatctataca ttaagacgac tcgaaatcca catatcaaat
     5521 atccgagtgt agtaaacatt ccaaaaccgt gatggaatgg aacaacactt aaaatcgcag
     5581 tatccggaat gatttgattg ccaaaaatag gatctctggc atgcgagaat ctgacgcagg
     5641 cagttctatg cggaagggcc acacccttag gtaacccagt agatccagag gaattcatta
     5701 tcagtgcaat tgttttgtca cgatcaaagg actctggtac aaaatcgtat tcattaaaac
     5761 cgggaggtag atgagatgtg acgaacgtgt acatcgactg aaatccctgg taatccgttt
     5821 tagaatccat gataataatt ttctggatta ttggtaattt tttttgcacg ttcaaaattt
     5881 tttgcaaccc ctttttggaa acaaacacta cggtaggctg cgaaatgttc atactgttga
     5941 gcaattcacg ttcattataa atgtcgttcg cgggcgcaac tgcaactccg ataaataacg
     6001 cgcccaacac cggcataaag aattgaagag agttttcact gcatacgacg attctgtgat
     6061 ttgtattcag cccatatcgt ttcatagctt ctgccaaccg aacggacatt tcgaagtatt
     6121 ccgcgtacgt gatgttcacc tcgatatgtg catctgtaaa agcaattgtt ccaggaacca
     6181 gggcgtatct cttcatagcc ttatgcagtt gctctccagc ggttccatcc tctagaggat
     6241 agaatggcgc cgggcctttc tttatgtttt tggcgtcctc cattcctctc caaatgaaat
     6301 gaacttcctt atatagagga agggtcttgc ggagacgtat aaacgcagaa aggcccaccc
     6361 gaaggtgagc cagtgtgact ctagtagaga gcgttcaccg acaaacaaca gataaaacga
     6421 aaggcccagt ctttcgactg agcctttcgt tttatttgat gcctggctct agtagcgatc
     6481 tacactagca ctatcagcgt tattaagcac cggtggagtg acgaccttca gcacgttcgt
     6541 actgttcaac gatggtgtag tcctcgttgt gggaggtgat gtccagtttg atgtcggttt
     6601 tgtaagcacc cggcagctga accggttttt tagccatgta ggtggtttta acttcagcgt
     6661 cgtagtgacc accgtctttc agtttcagac gcattttgat ttcacctttc aaagcaccgt
     6721 cctccgggta catacgttcg gtggaagctt cccaacccat ggtttttttc tgcataaccg
     6781 gaccgtcgga cgggaagttg gtaccacgca gtttaacttt gtagatgaac tcaccgtctt
     6841 gcagggagga gtcctgggta acggtaacaa caccaccgtc ctcgaagttc ataacacgtt
     6901 cccatttgaa accttccggg aaggacagtt tcaggtagtc cgggatgtca gccgggtgtt
     6961 taacgtaagc tttggaaccg tactggaact gcggggacag gatgtcccaa gcgaacggca
     7021 gcggaccacc tttggtaact ttcagtttag cggtctgggt accttcgtac ggacgacctt
     7081 caccttcacc ttcgatttcg aactcgtgac cgttaacgga accttccata cgaactttga
     7141 aacgcatgaa ctctttgata acgtcctcgg aggaagccat ctagtatttc tcctctttct
     7201 ctagtatgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     7261 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     7321 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     7381 cgcggggaga ggcggtttgc gtattgcgtc tcgtactccg gatccgaatt cggcattgtc
     7441 ttcacaga
//
