LOCUS       sequence-318518-        7068 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 7068)
  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 7068)
  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: pEPYC1CB0353
FEATURES             Location/Qualifiers
     source          1..7068
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..632
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    724..748
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    1034..1089
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             1091..2740
                     /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(1157..1175)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     CDS             2804..2827
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      2838..3545
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_feature    3594..3618
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(3774..4922)
                     /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(5258..5846)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5338..5357)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6017..6808,6809..6877))
                     /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     6640..6659
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6878..6982)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 agcgggccgg gagggttcga gaaggggggg cacccccctt cggcgtgcgc ggtcacgcgc
       61 acagggcgca gccctggtta aaaacaaggt ttataaatat tggtttaaaa gcaggttaaa
      121 agacaggtta gcggtggccg aaaaacgggc ggaaaccctt gcaaatgctg gattttctgc
      181 ctgtggacag cccctcaaat gtcaataggt gcgcccctca tctgtcagca ctctgcccct
      241 caagtgtcaa ggatcgcgcc cctcatctgt cagtagtcgc gcccctcaag tgtcaatacc
      301 gcagggcact tatccccagg cttgtccaca tcatctgtgg gaaactcgcg taaaatcagg
      361 cgttttcgcc gatttgcgag gctggccagc tccacgtcgc cggccgaaat cgagcctgcc
      421 cctcatctgt caacgccgcg ccgggtgagt cggcccctca agtgtcaacg tccgcccctc
      481 atctgtcagt gagggccaag ttttccgcga ggtatccaca acgccggcgg ccgcggtgtc
      541 tcgcacacgg cttcgacggc gtttctggcg cgtttgcagg gccatagacg gccgccagcc
      601 cagcggcgag ggcaaccagc ccggtgagcg tcgcaaagga gatcctgatc tgactgatgg
      661 gctgcctgta tcgagtggtg attttgtgcc gagctgccgg tcggggagct gttggctggc
      721 tggtggcagg atatattgtg gtgtaaacaa attgacgctt agacaactta ataacacatt
      781 gcggacgttt ttaatgtact ggggtggatg cagtgggccc cactctgtga agacaatgcc
      841 gaattcggat ccggagggag gcgtgtcgtt ttagtgaggc tggagtttaa tgagctaagc
      901 acatacgtca gaaaccatta ttgcgcgttc aaaagtcgcc taaggtcact atcagctagc
      961 aaatatctat ataaggtttt gctattcatt gaaagcagta gtgactgatt tgtatatata
     1021 ctgtattttt acaacaatta ccaacaacaa caaacaacaa acaacattac aattactatt
     1081 tacaattaca atggaggacg ccaaaaacat aaagaaaggc ccggcgccat tctatccgct
     1141 ggaagatgga accgctggag agcaactgca taaggctatg aagagatacg ccctggttcc
     1201 tggaacaatt gcttttacag atgcacatat cgaggtggac atcacttacg ctgagtactt
     1261 cgaaatgtcc gttcggttgg cagaagctat gaaacgatat gggctgaata caaatcacag
     1321 aatcgtcgta tgcagtgaaa actctcttca attctttatg ccggtgttgg gcgcgttatt
     1381 tatcggagtt gcagttgcgc ccgcgaacga catttataat gaacgtgaat tgctcaacag
     1441 tatgggcatt tcgcagccta ccgtggtgtt cgtttccaaa aaggggttgc aaaaaatttt
     1501 gaacgtgcaa aaaaagctcc caatcatcca aaaaattatt atcatggatt ctaaaacgga
     1561 ttaccaggga tttcagtcga tgtacacgtt cgtcacatct catctacctc ccggttttaa
     1621 tgaatacgat tttgtgccag agtccttcga tagggacaag acaattgcac tgatcatgaa
     1681 ctcctctgga tctactggtc tgcctaaagg tgtcgctctg cctcatagaa ctgcctgcgt
     1741 gagattctcg catgccagag atcctatttt tggcaatcaa atcattccgg atactgcgat
     1801 tttaagtgtt gttccattcc atcacggttt tggaatgttt actacactcg gatatttgat
     1861 atgtggattt cgagtcgtct taatgtatag atttgaagaa gaactgtttc tgaggagcct
     1921 tcaggattac aagattcaaa gtgcgctgct ggtgccaacc ctattctcct tcttcgccaa
     1981 aagcactctg attgacaaat acgatttatc taatttacac gaaattgctt ctggtggcgc
     2041 tcccctctct aaggaagtcg gggaagcggt tgccaagagg ttccatctgc caggtatcag
     2101 gcaaggatat gggctcactg agactacatc agctattctg attacacccg agggggatga
     2161 taaaccgggc gcggtcggta aagttgttcc attttttgaa gcgaaggttg tggatctgga
     2221 taccgggaaa acgctgggcg ttaatcaaag aggcgaactg tgtgtgagag gtcctatgat
     2281 tatgtccggt tatgtaaaca atccggaagc gaccaacgcc ttgattgaca aggatggatg
     2341 gctacattct ggagacatag cttactggga cgaggacgaa cacttcttca tcgttgaccg
     2401 cctgaagtct ctgattaagt acaaaggcta tcaggtggct cccgctgaat tggaatccat
     2461 cttgctccaa caccccaaca tcttcgacgc aggtgtcgca ggtctgcccg acgatgacgc
     2521 cggtgaactt cccgccgccg ttgttgtttt ggagcacgga aagacgatga cggaaaaaga
     2581 gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt
     2641 gtttgtggac gaagtaccga aaggtcttac cggaaaactc gacgcaagaa aaatcagaga
     2701 gatcctcata aaggccaaga agggcggaaa gatcgccgtg ggttcggatt ataaggacca
     2761 tgacggagac tataaggacc atgacctcga cgctgcagca gcggattata aggacgatga
     2821 cgataagtga gcttgtcctg ctttaatgag atatgcgaga agcctatgat cgcatgatat
     2881 ttgctttcaa ttctgttgtg cacgttgtaa aaaacctgag catgtgtagc tcagatcctt
     2941 accgccggtt tcggttcatt ctaatgaata tatcacccgt tactatcgta tttttatgaa
     3001 taatattctc cgttcaattt actgattgta ccctactact tatatgtaca atattaaaat
     3061 gaaaacaata tattgtgctg aataggttta tagcgacatc tatgatagag cgccacaata
     3121 acaaacaatt gcgttttatt attacaaatc caattttaaa aaaagcggca gaaccggtca
     3181 aacctaaaag actgattaca taaatcttat tcaaatttca aaagtgcccc aggggctagt
     3241 atctacgaca caccgagcgg cgaactaata acgctcactg aagggaactc cggttccccg
     3301 ccggcgcgca tgggtgagat tccttgaagt tgagtattgg ccgtccgctc taccgaaagt
     3361 tacgggcacc attcaacccg gtccagcacg gcggccgggt aaccgacttg ctgccccgag
     3421 aattatgcag catttttttg gtgtatgtgg gccccaaatg aagtgcaggt caaaccttga
     3481 cagtgacgac aaatcgttgg gcgggtccag ggcgaatttt gcgacaacat gtcgaggctc
     3541 agcaggaccg ctgcaattgt cttctgcacg aagtggttta aactatcagt gtttgacagg
     3601 atatattggc gggtaaacct aagagaaaag agcgtttatt agaataatcg gatatttaaa
     3661 agggcgtgaa aaggtttatc cgttcgtcca tttgtatgtg catgccaacc acagggttcc
     3721 ccagatcagg cgctggctgc tgaaccccca gccggaactg accccacaag gccctagcgt
     3781 ttgcaatgca ccaggtcatc attgacccag gcgtgttcca ccaggccgct gcctcgcaac
     3841 tcttcgcagg cttcgccgac ctgctcgcgc cacttcttca cgcgggtgga atccgatccg
     3901 cacatgaggc ggaaggtttc cagcttgagc gggtacggct cccggtgcga gctgaaatag
     3961 tcgaacatcc gtcgggccgt cggcgacagc ttgcggtact tctcccatat gaatttcgtg
     4021 tagtggtcgc cagcaaacag cacgacgatt tcctcgtcga tcaggacctg gcaacgggac
     4081 gttttcttgc cacggtccag gacgcggaag cggtgcagca gcgacaccga ttccaggtgc
     4141 ccaacgcggt cggacgtgaa gcccatcgcc gtcgcctgta ggcgcgacag gcattcctcg
     4201 gccttcgtgt aataccggcc attgatcgac cagcccaggt cctggcaaag ctcgtagaac
     4261 gtgaaggtga tcggctcgcc gataggggtg cgcttcgcgt actccaacac ctgctgccac
     4321 accagttcgt catcgtcggc ccgcagctcg acgccggtgt aggtgatctt cacgtccttg
     4381 ttgacgtgga aaatgacctt gttttgcagc gcctcgcgcg ggattttctt gttgcgcgtg
     4441 gtgaacaggg cagagcgggc cgtgtcgttt ggcatcgctc gcatcgtgtc cggccacggc
     4501 gcaatatcga acaaggaaag ctgcatttcc ttgatctgct gcttcgtgtg tttcagcaac
     4561 gcggcctgct tggcctcgct gacctgtttt gccaggtcct cgccggcggt ttttcgcttc
     4621 ttggtcgtca tagttcctcg cgtgtcgatg gtcatcgact tcgccaaacc tgccgcctcc
     4681 tgttcaagac gacgcgaacg ctccacggcg gccgatggcg cgggcagggc agggggagcc
     4741 agttgcacgc tgtcgcgctc gatcttggcc gtagcttgct ggaccatcga gccgacggac
     4801 tggaaggttt cgcggggcgc acgcatgacg gtgcggcttg cgatggtttc ggcatcctcg
     4861 gcggaaaacc ccgcgtcgat cagttcttgc ctgtatgcct tccggtcaaa cgtccgattc
     4921 attcaccctc cttgcgggat tgccccgact cacgccgggg caatgtgccc ttattcctga
     4981 tttgacccgc ctggtgcctt ggtgtccaga taatccacct tatcggcaat gaagtcggtc
     5041 ccgtagaccg tctggccgtc cttctcgtac ttggtattcc gaatcttgcc ctgcacgaat
     5101 accagcgacc ccttgcccaa atacttgccg tgggcctcgg cctgagagcc aaaacacttg
     5161 atgcggaaga agtcggtgcg ctcctgcttg tcgccggcat cgttgcgcca catctaggat
     5221 ctgccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5281 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5341 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5401 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5461 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5521 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5581 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5641 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5701 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5761 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5821 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5881 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5941 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6001 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6061 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6121 gcttaccatc tggccccagt gctgcaatga taccgcgaga accacgctca ccggctccag
     6181 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6241 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6301 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6361 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6421 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6481 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6541 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6601 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6661 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6721 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6781 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6841 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6901 gaagcattta tcaaggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6961 ataaacaaat aggggttccg cgcacgaatt ggccagcgct gccatttttg gggtgaggcc
     7021 gttcgcggcc gaggggcgca gcccctgggg ggatgggagg cccgcgtt
//
