LOCUS       sequence-486338-        3813 bp    DNA     circular SYN 16-SEP-2025
DEFINITION  Firefly luciferase plus as a Level-0 part (Golden Gate).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3813)
  AUTHORS   Khan MA, Herring G, Zhu JY, Oliva M, Fourie E, Johnston B, Zhang Z, 
            Potter J, Pineda L, Pflueger J, Swain T, Pflueger C, Lloyd JPB, 
            Secco D, Small I, Kidd BN, Lister R
  TITLE     CRISPRi-based circuits to control gene expression in plants.
  JOURNAL   Nat Biotechnol. 2025 Mar;43(3):416-430. doi: 
            10.1038/s41587-024-02236-w. Epub 2024 May 20.
  PUBMED    38769424
REFERENCE   2  (bases 1 to 3813)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41587-024-02236-w"; journalName: "Nat Biotechnol"; date: 
            "2025-03"; volume: "43"; issue: "3"; pages: "416-430"
COMMENT     Sequence Label: LLP1022
FEATURES             Location/Qualifiers
     source          1..3813
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             285..1076
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      1417..2005
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1906..1925
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2058..3707
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(2124..2142)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
ORIGIN
        1 gtgccgaggc acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt
       61 agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg
      121 cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg
      181 catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg
      241 atgttacgca gcagggcagt cgccctaaaa caaagttaaa catcatgggg gaagcggtga
      301 tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac
      361 cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca
      421 gcgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt
      481 tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg
      541 tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg
      601 aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca
      661 cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg
      721 taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc
      781 taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg
      841 tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg
      901 atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg
      961 aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt
     1021 tggaagaatt tgtccattac gtgaaaggcg agatcaccaa ggtagtcggc aaataatgtc
     1081 tagctagaaa ttcgttcaag ccgacgccgc ttcgcggcgc ggcttaactc aagcgttaga
     1141 tgcactaagc acataattgc tcacagccaa actatcaggt caagtctgct tttattattt
     1201 ttaagcgtgc ataataagcc ctacacaaat tgggagatat atcatgctgt cagaccaagt
     1261 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     1321 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     1381 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     1441 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     1501 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     1561 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     1621 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     1681 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     1741 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     1801 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     1861 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     1921 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     1981 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctgca
     2041 ctctgtggtc tcaaatgatg gaggacgcca aaaacataaa gaaaggcccg gcgccattct
     2101 atccgctgga agatggaacc gctggagagc aactgcataa ggctatgaag agatacgccc
     2161 tggttcctgg aacaattgct tttacagatg cacatatcga ggtggacatc acttacgctg
     2221 agtacttcga aatgtccgtt cggttggcag aagctatgaa acgatatggg ctgaatacaa
     2281 atcacagaat cgtcgtatgc agtgaaaact ctcttcaatt ctttatgccg gtgttgggcg
     2341 cgttatttat cggagttgca gttgcgcccg cgaacgacat ttataatgaa cgtgaattgc
     2401 tcaacagtat gggcatttcg cagcctaccg tggtgttcgt ttccaaaaag gggttgcaaa
     2461 aaattttgaa cgtgcaaaaa aagctcccaa tcatccaaaa aattattatc atggattcta
     2521 aaacggatta ccagggattt cagtcgatgt acacgttcgt cacatctcat ctacctcccg
     2581 gttttaatga atacgatttt gtgccagagt ccttcgatag ggacaagaca attgcactga
     2641 tcatgaactc ctctggatct actggtctgc ctaaaggtgt cgctctgcct catagaactg
     2701 cctgcgtgag attctcgcat gccagagatc ctatttttgg caatcaaatc attccggata
     2761 ctgcgatttt aagtgttgtt ccattccatc acggttttgg aatgtttact acactcggat
     2821 atttgatatg tggatttcga gtcgtcttaa tgtatagatt tgaagaggag ctgtttctga
     2881 ggagccttca ggattacaag attcaaagtg cgctgctggt gccaacccta ttctccttct
     2941 tcgccaaaag cactctgatt gacaaatacg atttatctaa tttacacgaa attgcttctg
     3001 gtggcgctcc cctctctaag gaagtcgggg aagcggttgc caagaggttc catctgccag
     3061 gtatcaggca aggatatggg ctcactgaga ctacatcagc tattctgatt acacccgagg
     3121 gggatgataa accgggcgcg gtcggtaaag ttgttccatt ttttgaagcg aaggttgtgg
     3181 atctggatac cgggaaaacg ctgggcgtta atcaaagagg cgaactgtgt gtgagaggtc
     3241 ctatgattat gtccggttat gtaaacaatc cggaagcgac caacgccttg attgacaagg
     3301 atggatggct acattctgga gacatagctt actgggacga ggacgaacac ttcttcatcg
     3361 ttgaccgcct gaagtctctg attaagtaca aaggctatca ggtggctccc gctgaattgg
     3421 aatccatctt gctccaacac cccaacatct tcgacgcagg tgtcgcaggt ctacccgacg
     3481 atgacgccgg tgaacttccc gccgccgttg ttgttttgga gcacggaaag acgatgacgg
     3541 aaaaagagat cgtggattac gtcgccagtc aagtaacaac cgcgaaaaag ttgcgcggag
     3601 gagttgtgtt tgtggacgaa gtaccgaaag gtcttaccgg aaaactcgac gcaagaaaaa
     3661 tcagagagat cctcataaag gccaagaagg gcggaaagat cgccgtgggt tctcgatctg
     3721 atgcttctga agtaacgtca acaagctccc agtctgaggt atgcacagtg gaaacgccag
     3781 gttgtgttgg ttcttgagct tcgagaccac gaa
//
