LOCUS       sequence-308058-        6380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luminescent reporter for cytokinin signaling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6380)
  AUTHORS   Lehmann S, Dominguez-Ferreras A, Huang WJ, Denby K, Ntoukakis V, 
            Schafer P
  TITLE     Novel markers for high-throughput protoplast-based analyses of 
            phytohormone signaling.
  JOURNAL   PLoS One. 2020 Jun 4;15(6):e0234154. doi: 
            10.1371/journal.pone.0234154. eCollection 2020.
  PUBMED    32497144
REFERENCE   2  (bases 1 to 6380)
  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: "PLoS One.";
            date: "2020-06-4"; volume: "15(6):e0234154. doi"; pages: " 
            10.1371/journal.pone.0234154. eCollection 2020"
COMMENT     Sequence Label: pARR5::LUC
FEATURES             Location/Qualifiers
     source          1..6380
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(14..30)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14..30)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(27..49)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    38..54
                     /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(62..68,69..86,87..92))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(245..262)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(416..1004)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(496..515)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1175..1966,1967..2035))
                     /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     1798..1817
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2036..2140)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2208..2226
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     2386..2405
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2599..2621
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2613..2630
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2614..2630
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4316..5968
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(4382..4400)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     terminator      6123..6375
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
ORIGIN
        1 aattcgtaat catgtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
       61 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
      121 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
      181 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
      241 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
      301 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
      361 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
      421 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
      481 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
      541 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
      601 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
      661 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
      721 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
      781 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
      841 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
      901 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
      961 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     1021 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     1081 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     1141 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     1201 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     1261 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     1321 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     1381 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     1441 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     1501 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     1561 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     1621 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     1681 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     1741 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     1801 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     1861 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     1921 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     1981 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     2041 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     2101 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     2161 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     2221 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     2281 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     2341 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     2401 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     2461 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     2521 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     2581 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gccaagcttg
     2641 catgcctgca ggctctagag gatccccaat aaagcatatt tgtcttcagc ctctcatact
     2701 atttgagagg taaaaaccga gaccattagg attattcttt atagaatgtt ttggtgccta
     2761 acaaatttta ctggcatttc ttttttttgc cataaactgt taagaggttt atttatgaat
     2821 tgttaactta atttctttga tcaaacaata gtaagttcta taaatgtatc ttgtgatctt
     2881 tgactatgat agacactaag ctcggactca tgtacatagc caaaacactc atatatatta
     2941 aaacaataat ttcatcggta aactcatgtt gtttccaaat atacaagtag agaaaacgtg
     3001 tagacttact ctgaagggac acgcatatac atactgataa acaaccatca tggagaaatc
     3061 cgagccgttt ctctgcaagc caaaccaaat ttgacatata tagtaaggtc atgtgctcta
     3121 tacacctagt acctcacatt catttgatat ctcctctaga gatacatcga tcatagtcat
     3181 attttcagct ttcaatgatg atttagtcat taccattctt gaaatatatt ggtcgagttt
     3241 agtccataga tcttcgatcc tgtcgtgaca aagccttcca tcaattgtca aaagatctct
     3301 cacgtgtgga aatactcgat gcgtgttgct aaatatgtga ataaaagaac gccaaattta
     3361 tagaaaaagc gaattaaaat tcaagtaaac atatgaatat ttttcaagaa aagtgaattg
     3421 aaattttatt ttcatgaaaa cttactattt tttttttttt tttgaacact catgaaaact
     3481 tactattaac tcatcaatct tatgccattc gaccacgatc gatagtagag gcaactatta
     3541 tggtacagtt acgagatgga caaacaacaa taacttgaaa aataatgatt ttttgttttg
     3601 tttctagaaa tatacacata cggatcttga atttatctga ttgcgcaact agagatatca
     3661 acattgaaat tcattcgccc aaatagtcaa tctggcggtc acatgtcagc taacttttcc
     3721 aacaatgttc attaattaat taacaatctc aaagattttg tagattgaaa tacaaatctt
     3781 ctctctgtgg tacatttctt gaaaaatggg aaaatcaaga aagtatcgaa aatgtacaaa
     3841 aataaaaaga aatgaatcaa agtagccatg atcttgacaa caataatcga gagagatcgt
     3901 catgatacga tttccctcat ccaaaattga ttttatttcc cttcccaaat caaacatatc
     3961 atatgatttc accactcacc attacttgac tattctcaac aaaaaaaata ttaaaaaact
     4021 ttatgacttt gattttattt ttatttgaag tttagccaaa atttgaaaat atgacttttg
     4081 agaagaaaac agaataaaca aataattagc cacgcgctat cagacagaca aaatcccaca
     4141 gatatgcaaa gatctctcag aatcctctcc ccatatcata tttttctctt ttccctctcc
     4201 ttctttcttc ctttataaat ccatttattc tcctctcatc tctcagcaaa atcaaatcct
     4261 catagttgat tctctctatc tctctcacga gtcacgatcc tactcttctt gatccatgga
     4321 agacgccaaa aacataaaga aaggcccggc gccattctat cctctagagg atggaaccgc
     4381 tggagagcaa ctgcataagg ctatgaagag atacgccctg gttcctggaa caattgcttt
     4441 tacagatgca catatcgagg tgaacatcac gtacgcggaa tacttcgaaa tgtccgttcg
     4501 gttggcagaa gctatgaaac gatatgggct gaatacaaat cacagaatcg tcgtatgcag
     4561 tgaaaactct cttcaattct ttatgccggt gttgggcgcg ttatttatcg gagttgcagt
     4621 tgcgcccgcg aacgacattt ataatgaacg tgaattgctc aacagtatga acatttcgca
     4681 gcctaccgta gtgtttgttt ccaaaaaggg gttgcaaaaa attttgaacg tgcaaaaaaa
     4741 attaccaata atccagaaaa ttattatcat ggattctaaa acggattacc agggatttca
     4801 gtcgatgtac acgttcgtca catctcatct acctcccggt tttaatgaat acgattttgt
     4861 accagagtcc tttgatcgtg acaaaacaat tgcactgata atgaattcct ctggatctac
     4921 tgggttacct aagggtgtgg cccttccgca tagaactgcc tgcgtcagat tctcgcatgc
     4981 cagagatcct atttttggca atcaaatcat tccggatact gcgattttaa gtgttgttcc
     5041 attccatcac ggttttggaa tgtttactac actcggatat ttgatatgtg gatttcgagt
     5101 cgtcttaatg tatagatttg aagaagagct gtttttacga tcccttcagg attacaaaat
     5161 tcaaagtgcg ttgctagtac caaccctatt ttcattcttc gccaaaagca ctctgattga
     5221 caaatacgat ttatctaatt tacacgaaat tgcttctggg ggcgcacctc tttcgaaaga
     5281 agtcggggaa gcggttgcaa aacgcttcca tcttccaggg atacgacaag gatatgggct
     5341 cactgagact acatcagcta ttctgattac acccgagggg gatgataaac cgggcgcggt
     5401 cggtaaagtt gttccatttt ttgaagcgaa ggttgtggat ctggataccg ggaaaacgct
     5461 gggcgttaat cagagaggcg aattatgtgt cagaggacct atgattatgt ccggttatgt
     5521 aaacaatccg gaagcgacca acgccttgat tgacaaggat ggatggctac attctggaga
     5581 catagcttac tgggacgaag acgaacactt cttcatagtt gaccgcttga agtctttaat
     5641 taaatacaaa ggatatcagg tggcccccgc tgaattggaa tcgatattgt tacaacaccc
     5701 caacatcttc gacgcgggcg tggcaggtct tcccgacgat gacgccggtg aacttcccgc
     5761 cgccgttgtt gttttggagc acggaaagac gatgacggaa aaagagatcg tggattacgt
     5821 cgccagtcaa gtaacaaccg cgaaaaagtt gcgcggagga gttgtgtttg tggacgaagt
     5881 accgaaaggt cttaccggaa aactcgacgc aagaaaaatc agagagatcc tcataaaggc
     5941 caagaagggc ggaaagtcca aattgtaaaa tgtaactgta ttcagcgatg acgaaattct
     6001 tagctattgt aatattatat gcaaattgat gaatggtaat tttgtaattg tgggtcactg
     6061 tactatttta acgaataata aaatcaggta taggtaacta aaaaggaatt cgagctcgaa
     6121 ttgatcgttc aaacatttgg caataaagtt tcttaagatt gaatcctgtt gccggtcttg
     6181 cgatgattat catataattt ctgttgaatt acgttaagca tgtaataatt aacatgtaat
     6241 gcatgacgtt atttatgaga tgggttttta tgattagagt cccgcaatta tacatttaat
     6301 acgcgataga aaacaaaata tagcgcgcaa actaggataa attatcgcgc gcggtgtcat
     6361 ctatgttact agatctctag
//
