LOCUS       sequence-342374-        5591 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1 luciferase reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5591)
  AUTHORS   Cai YM, Kallam K, Tidd H, Gendarini G, Salzman A, Patron NJ
  TITLE     Rational design of minimal synthetic promoters for plants.
  JOURNAL   Nucleic Acids Res. 2020 Aug 28. pii: 5897334. doi: 
            10.1093/nar/gkaa682.
  PUBMED    32856047
REFERENCE   2  (bases 1 to 5591)
  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: "Nucleic 
            Acids Res. 2020 Aug 28. pii: 5897334. doi: 10.1093/nar/gkaa682."
COMMENT     Sequence Label: pEPYC1CB0197
FEATURES             Location/Qualifiers
     source          1..5591
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..25
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        210..389
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    425..480
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             482..994
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             1055..1078
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      1094..1346
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    1394..1418
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(1574..2722)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      complement(3058..3646)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3138..3157)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3817..4608,4609..4677))
                     /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     4440..4459
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4678..4782)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4869..5500
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 tggcaggata tattgtggtg taaacaaatt gacgcttaga caacttaata acacattgcg
       61 gacgttttta atgtactggg gtggatgcag tgggccccac tctgtgaaga caagcaagaa
      121 ttcaagcttg gagagcggag aattaaggga gtcacgttat gacccccgcc gatgacgcgg
      181 gacaagccgt tttacgtttg gaactgacag aaccgcaacg ttgaaggagc cactcagccg
      241 cgggtttctg gagtttaatg agctaagcac atacgtcaga aaccattatt gcgcgttcaa
      301 aagtcgccta aggtcactat cagctagcaa atatttcttg tcaaaaatgc tccactgacg
      361 ttccataaat tcccctcggt atccaattag agtctcatat tcactctcat actgtatttt
      421 tacaacaatt accaacaaca acaaacaaca aacaacatta caattactat ttacaattac
      481 aatggtattc acactcgaag atttcgttgg ggactggcga cagacagccg gctacaacct
      541 ggaccaagtc cttgaacagg gaggtgtgtc cagtttgttt cagaatctcg gggtgtccgt
      601 aactccgatc caaaggattg tcctgagcgg tgaaaatggg ctgaagatcg acatccatgt
      661 catcatcccg tatgaaggtc tgagcggcga ccaaatgggc cagatcgaaa aaatttttaa
      721 ggtggtgtac cctgtggatg atcatcactt taaggtgatc ctgcactatg gcacactggt
      781 aatcgacggg gttacgccga acatgatcga ctatttcgga cggccgtatg aaggcatcgc
      841 cgtgttcgac ggcaaaaaga tcactgtaac agggaccctg tggaacggca acaaaattat
      901 cgacgagcgc ctgatcaacc ccgacggctc cctgctgttc cgagtaacca tcaacggagt
      961 gaccggctgg cggctgtgcg aacgcattct ggcttcggat tataaggacc atgacggaga
     1021 ctataaggac catgacctcg acgctgcagc agcggattat aaggacgatg acgataagtg
     1081 agcttgtcaa gcagatcgtt caaacatttg gcaataaagt ttcttaagat tgaatcctgt
     1141 tgccggtctt gcgatgatta tcatataatt tctgttgaat tacgttaagc atgtaataat
     1201 taacatgtaa tgcatgacgt tatttatgag atgggttttt atgattagag tcccgcaatt
     1261 atacatttaa tacgcgatag aaaacaaaat atagcgcgca aactaggata aattatcgcg
     1321 cgcggtgtca tctatgttac tagatcgacg ctactattgt cttctgcacg aagtggttta
     1381 aactatcagt gtttgacagg atatattggc gggtaaacct aagagaaaag agcgtttatt
     1441 agaataatcg gatatttaaa agggcgtgaa aaggtttatc cgttcgtcca tttgtatgtg
     1501 catgccaacc acagggttcc ccagatcagg cgctggctgc tgaaccccca gccggaactg
     1561 accccacaag gccctagcgt ttgcaatgca ccaggtcatc attgacccag gcgtgttcca
     1621 ccaggccgct gcctcgcaac tcttcgcagg cttcgccgac ctgctcgcgc cacttcttca
     1681 cgcgggtgga atccgatccg cacatgaggc ggaaggtttc cagcttgagc gggtacggct
     1741 cccggtgcga gctgaaatag tcgaacatcc gtcgggccgt cggcgacagc ttgcggtact
     1801 tctcccatat gaatttcgtg tagtggtcgc cagcaaacag cacgacgatt tcctcgtcga
     1861 tcaggacctg gcaacgggac gttttcttgc cacggtccag gacgcggaag cggtgcagca
     1921 gcgacaccga ttccaggtgc ccaacgcggt cggacgtgaa gcccatcgcc gtcgcctgta
     1981 ggcgcgacag gcattcctcg gccttcgtgt aataccggcc attgatcgac cagcccaggt
     2041 cctggcaaag ctcgtagaac gtgaaggtga tcggctcgcc gataggggtg cgcttcgcgt
     2101 actccaacac ctgctgccac accagttcgt catcgtcggc ccgcagctcg acgccggtgt
     2161 aggtgatctt cacgtccttg ttgacgtgga aaatgacctt gttttgcagc gcctcgcgcg
     2221 ggattttctt gttgcgcgtg gtgaacaggg cagagcgggc cgtgtcgttt ggcatcgctc
     2281 gcatcgtgtc cggccacggc gcaatatcga acaaggaaag ctgcatttcc ttgatctgct
     2341 gcttcgtgtg tttcagcaac gcggcctgct tggcctcgct gacctgtttt gccaggtcct
     2401 cgccggcggt ttttcgcttc ttggtcgtca tagttcctcg cgtgtcgatg gtcatcgact
     2461 tcgccaaacc tgccgcctcc tgttcaagac gacgcgaacg ctccacggcg gccgatggcg
     2521 cgggcagggc agggggagcc agttgcacgc tgtcgcgctc gatcttggcc gtagcttgct
     2581 ggaccatcga gccgacggac tggaaggttt cgcggggcgc acgcatgacg gtgcggcttg
     2641 cgatggtttc ggcatcctcg gcggaaaacc ccgcgtcgat cagttcttgc ctgtatgcct
     2701 tccggtcaaa cgtccgattc attcaccctc cttgcgggat tgccccgact cacgccgggg
     2761 caatgtgccc ttattcctga tttgacccgc ctggtgcctt ggtgtccaga taatccacct
     2821 tatcggcaat gaagtcggtc ccgtagaccg tctggccgtc cttctcgtac ttggtattcc
     2881 gaatcttgcc ctgcacgaat accagcgacc ccttgcccaa atacttgccg tgggcctcgg
     2941 cctgagagcc aaaacacttg atgcggaaga agtcggtgcg ctcctgcttg tcgccggcat
     3001 cgttgcgcca catctaggat ctgccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3061 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3121 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3181 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3241 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3301 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3361 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3421 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3481 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3541 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3601 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3661 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3721 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3781 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     3841 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     3901 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     3961 accacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4021 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4081 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4141 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4201 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4261 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4321 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4381 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4441 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4501 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4561 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4621 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4681 cttccttttt caatattatt gaagcattta tcaaggttat tgtctcatga gcggatacat
     4741 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacgaatt ggccagcgct
     4801 gccatttttg gggtgaggcc gttcgcggcc gaggggcgca gcccctgggg ggatgggagg
     4861 cccgcgttag cgggccggga gggttcgaga agggggggca ccccccttcg gcgtgcgcgg
     4921 tcacgcgcac agggcgcagc cctggttaaa aacaaggttt ataaatattg gtttaaaagc
     4981 aggttaaaag acaggttagc ggtggccgaa aaacgggcgg aaacccttgc aaatgctgga
     5041 ttttctgcct gtggacagcc cctcaaatgt caataggtgc gcccctcatc tgtcagcact
     5101 ctgcccctca agtgtcaagg atcgcgcccc tcatctgtca gtagtcgcgc ccctcaagtg
     5161 tcaataccgc agggcactta tccccaggct tgtccacatc atctgtggga aactcgcgta
     5221 aaatcaggcg ttttcgccga tttgcgaggc tggccagctc cacgtcgccg gccgaaatcg
     5281 agcctgcccc tcatctgtca acgccgcgcc gggtgagtcg gcccctcaag tgtcaacgtc
     5341 cgcccctcat ctgtcagtga gggccaagtt ttccgcgagg tatccacaac gccggcggcc
     5401 gcggtgtctc gcacacggct tcgacggcgt ttctggcgcg tttgcagggc catagacggc
     5461 cgccagccca gcggcgaggg caaccagccc ggtgagcgtc gcaaaggaga tcctgatctg
     5521 actgatgggc tgcctgtatc gagtggtgat tttgtgccga gctgccggtc ggggagctgt
     5581 tggctggctg g
//
