LOCUS       sequence-308023-        5654 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Luminescent reporter for auxin signaling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5654)
  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 5654)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pIAA5::LUC
FEATURES             Location/Qualifiers
     source          1..5654
                     /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             3590..5242
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(3656..3674)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     terminator      5397..5649
                     /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 gatccgctgt ccattatcac aaagtcttgt tgtgttattc
     2701 attggctaaa gccagccgac aatatattat acgcaattct acatgatcaa tcgaaaacta
     2761 gaaattgtgt acaaggaaat ttgactaatc tagcttatct atcaacatgt gaacattcca
     2821 aaaatgtact attggagata caaagatatg gaaatatcta tgaaaaatgg aaatatatcc
     2881 cgtaattgat attttgatca gttctgtttc tatttcgtaa caaataatga caaaagtatg
     2941 gtgtacatgt gtagattacg tatgatgtaa tgaaagtatc gggcacatgt gtgaatagca
     3001 gaaacaagat atatgacgaa aacgtagatg catttttttc tttgaaatta tgtatgcact
     3061 gtgtagtgtg ttacgtgtag aaaaaaatga aataatataa taacttgtct tagccttgag
     3121 taacacgtgt tcggttgtcg agaaacataa atatacgtag attttaaagt gcactaatat
     3181 atattcgacc atgaatatgt tgttttatta ttatgtcgtt gattaaaaga atgggataca
     3241 gacataacaa atcacacata acatgggatg tctaatagaa agtaacgtgt ttctgatcca
     3301 acggctgaga aagaaaccct actttgtcgg taacaaaatc gtggccgtcc agtgtgtcga
     3361 accaacggat ccatattatc gacatttaaa cacgaagcgg caaaggaatg aaaaaacatg
     3421 acataaaaat ttcgacaaga tcttcgaatg gggccgacaa gataaatcaa tgctaaacaa
     3481 gcttgtctca aacgccatcg tttatgttcc catgagaaca tataaagtgg caacagaata
     3541 aagttcaaac acattaacac caatactttc aatcaaaaac atcaaagcca tggaagacgc
     3601 caaaaacata aagaaaggcc cggcgccatt ctatcctcta gaggatggaa ccgctggaga
     3661 gcaactgcat aaggctatga agagatacgc cctggttcct ggaacaattg cttttacaga
     3721 tgcacatatc gaggtgaaca tcacgtacgc ggaatacttc gaaatgtccg ttcggttggc
     3781 agaagctatg aaacgatatg ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa
     3841 ctctcttcaa ttctttatgc cggtgttggg cgcgttattt atcggagttg cagttgcgcc
     3901 cgcgaacgac atttataatg aacgtgaatt gctcaacagt atgaacattt cgcagcctac
     3961 cgtagtgttt gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa aaaaattacc
     4021 aataatccag aaaattatta tcatggattc taaaacggat taccagggat ttcagtcgat
     4081 gtacacgttc gtcacatctc atctacctcc cggttttaat gaatacgatt ttgtaccaga
     4141 gtcctttgat cgtgacaaaa caattgcact gataatgaat tcctctggat ctactgggtt
     4201 acctaagggt gtggcccttc cgcatagaac tgcctgcgtc agattctcgc atgccagaga
     4261 tcctattttt ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg ttccattcca
     4321 tcacggtttt ggaatgttta ctacactcgg atatttgata tgtggatttc gagtcgtctt
     4381 aatgtataga tttgaagaag agctgttttt acgatccctt caggattaca aaattcaaag
     4441 tgcgttgcta gtaccaaccc tattttcatt cttcgccaaa agcactctga ttgacaaata
     4501 cgatttatct aatttacacg aaattgcttc tgggggcgca cctctttcga aagaagtcgg
     4561 ggaagcggtt gcaaaacgct tccatcttcc agggatacga caaggatatg ggctcactga
     4621 gactacatca gctattctga ttacacccga gggggatgat aaaccgggcg cggtcggtaa
     4681 agttgttcca ttttttgaag cgaaggttgt ggatctggat accgggaaaa cgctgggcgt
     4741 taatcagaga ggcgaattat gtgtcagagg acctatgatt atgtccggtt atgtaaacaa
     4801 tccggaagcg accaacgcct tgattgacaa ggatggatgg ctacattctg gagacatagc
     4861 ttactgggac gaagacgaac acttcttcat agttgaccgc ttgaagtctt taattaaata
     4921 caaaggatat caggtggccc ccgctgaatt ggaatcgata ttgttacaac accccaacat
     4981 cttcgacgcg ggcgtggcag gtcttcccga cgatgacgcc ggtgaacttc ccgccgccgt
     5041 tgttgttttg gagcacggaa agacgatgac ggaaaaagag atcgtggatt acgtcgccag
     5101 tcaagtaaca accgcgaaaa agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa
     5161 aggtcttacc ggaaaactcg acgcaagaaa aatcagagag atcctcataa aggccaagaa
     5221 gggcggaaag tccaaattgt aaaatgtaac tgtattcagc gatgacgaaa ttcttagcta
     5281 ttgtaatatt atatgcaaat tgatgaatgg taattttgta attgtgggtc actgtactat
     5341 tttaacgaat aataaaatca ggtataggta actaaaaagg aattcgagct cgaattgatc
     5401 gttcaaacat ttggcaataa agtttcttaa gattgaatcc tgttgccggt cttgcgatga
     5461 ttatcatata atttctgttg aattacgtta agcatgtaat aattaacatg taatgcatga
     5521 cgttatttat gagatgggtt tttatgatta gagtcccgca attatacatt taatacgcga
     5581 tagaaaacaa aatatagcgc gcaaactagg ataaattatc gcgcgcggtg tcatctatgt
     5641 tactagatct ctag
//
