LOCUS       sequence-318554-        7339 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 7339)
  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 7339)
  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: pEPYC1CB0334
FEATURES             Location/Qualifiers
     source          1..7339
                     /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    1305..1360
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             1362..3011
                     /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(1428..1446)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     CDS             3075..3098
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      3109..3816
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_feature    3865..3889
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(4045..5193)
                     /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(5529..6117)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5609..5628)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6288..7079,7080..7148))
                     /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     6911..6930
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7149..7253)
                     /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 ccggagtgga agattcagaa aatcgattaa tggattgatc aacatcctta
      901 ccgctatggg taagattgat gaaaagtcaa aaacaaaaat caattatgca caccagcatg
      961 tgttgatcac cagctattgt gggacaccaa tttcgtccac agacatcaac atcttatcgt
     1021 cctttgaaga taagataata atgttgaaga taagagtggg agccaccact aaaacattgc
     1081 tttgtcaaaa gctaaaaaag atgatgcccg acagccactt gtgtgaagca tgtgaagccg
     1141 gtccctccac taagaaaatt agtgaagcat cttccagtgg tccctccact cacagctcaa
     1201 tcagtgagca acaggacgaa ggaaatgacg taagccatga cgtctaatcc cacaagaatt
     1261 tccttatata aggaacacaa atcagaaggt actgtatttt tacaacaatt accaacaaca
     1321 acaaacaaca aacaacatta caattactat ttacaattac aatggaggac gccaaaaaca
     1381 taaagaaagg cccggcgcca ttctatccgc tggaagatgg aaccgctgga gagcaactgc
     1441 ataaggctat gaagagatac gccctggttc ctggaacaat tgcttttaca gatgcacata
     1501 tcgaggtgga catcacttac gctgagtact tcgaaatgtc cgttcggttg gcagaagcta
     1561 tgaaacgata tgggctgaat acaaatcaca gaatcgtcgt atgcagtgaa aactctcttc
     1621 aattctttat gccggtgttg ggcgcgttat ttatcggagt tgcagttgcg cccgcgaacg
     1681 acatttataa tgaacgtgaa ttgctcaaca gtatgggcat ttcgcagcct accgtggtgt
     1741 tcgtttccaa aaaggggttg caaaaaattt tgaacgtgca aaaaaagctc ccaatcatcc
     1801 aaaaaattat tatcatggat tctaaaacgg attaccaggg atttcagtcg atgtacacgt
     1861 tcgtcacatc tcatctacct cccggtttta atgaatacga ttttgtgcca gagtccttcg
     1921 atagggacaa gacaattgca ctgatcatga actcctctgg atctactggt ctgcctaaag
     1981 gtgtcgctct gcctcataga actgcctgcg tgagattctc gcatgccaga gatcctattt
     2041 ttggcaatca aatcattccg gatactgcga ttttaagtgt tgttccattc catcacggtt
     2101 ttggaatgtt tactacactc ggatatttga tatgtggatt tcgagtcgtc ttaatgtata
     2161 gatttgaaga agaactgttt ctgaggagcc ttcaggatta caagattcaa agtgcgctgc
     2221 tggtgccaac cctattctcc ttcttcgcca aaagcactct gattgacaaa tacgatttat
     2281 ctaatttaca cgaaattgct tctggtggcg ctcccctctc taaggaagtc ggggaagcgg
     2341 ttgccaagag gttccatctg ccaggtatca ggcaaggata tgggctcact gagactacat
     2401 cagctattct gattacaccc gagggggatg ataaaccggg cgcggtcggt aaagttgttc
     2461 cattttttga agcgaaggtt gtggatctgg ataccgggaa aacgctgggc gttaatcaaa
     2521 gaggcgaact gtgtgtgaga ggtcctatga ttatgtccgg ttatgtaaac aatccggaag
     2581 cgaccaacgc cttgattgac aaggatggat ggctacattc tggagacata gcttactggg
     2641 acgaggacga acacttcttc atcgttgacc gcctgaagtc tctgattaag tacaaaggct
     2701 atcaggtggc tcccgctgaa ttggaatcca tcttgctcca acaccccaac atcttcgacg
     2761 caggtgtcgc aggtctgccc gacgatgacg ccggtgaact tcccgccgcc gttgttgttt
     2821 tggagcacgg aaagacgatg acggaaaaag agatcgtgga ttacgtcgcc agtcaagtaa
     2881 caaccgcgaa aaagttgcgc ggaggagttg tgtttgtgga cgaagtaccg aaaggtctta
     2941 ccggaaaact cgacgcaaga aaaatcagag agatcctcat aaaggccaag aagggcggaa
     3001 agatcgccgt gggttcggat tataaggacc atgacggaga ctataaggac catgacctcg
     3061 acgctgcagc agcggattat aaggacgatg acgataagtg agcttgtcct gctttaatga
     3121 gatatgcgag aagcctatga tcgcatgata tttgctttca attctgttgt gcacgttgta
     3181 aaaaacctga gcatgtgtag ctcagatcct taccgccggt ttcggttcat tctaatgaat
     3241 atatcacccg ttactatcgt atttttatga ataatattct ccgttcaatt tactgattgt
     3301 accctactac ttatatgtac aatattaaaa tgaaaacaat atattgtgct gaataggttt
     3361 atagcgacat ctatgataga gcgccacaat aacaaacaat tgcgttttat tattacaaat
     3421 ccaattttaa aaaaagcggc agaaccggtc aaacctaaaa gactgattac ataaatctta
     3481 ttcaaatttc aaaagtgccc caggggctag tatctacgac acaccgagcg gcgaactaat
     3541 aacgctcact gaagggaact ccggttcccc gccggcgcgc atgggtgaga ttccttgaag
     3601 ttgagtattg gccgtccgct ctaccgaaag ttacgggcac cattcaaccc ggtccagcac
     3661 ggcggccggg taaccgactt gctgccccga gaattatgca gcattttttt ggtgtatgtg
     3721 ggccccaaat gaagtgcagg tcaaaccttg acagtgacga caaatcgttg ggcgggtcca
     3781 gggcgaattt tgcgacaaca tgtcgaggct cagcaggacc gctgcaattg tcttctgcac
     3841 gaagtggttt aaactatcag tgtttgacag gatatattgg cgggtaaacc taagagaaaa
     3901 gagcgtttat tagaataatc ggatatttaa aagggcgtga aaaggtttat ccgttcgtcc
     3961 atttgtatgt gcatgccaac cacagggttc cccagatcag gcgctggctg ctgaaccccc
     4021 agccggaact gaccccacaa ggccctagcg tttgcaatgc accaggtcat cattgaccca
     4081 ggcgtgttcc accaggccgc tgcctcgcaa ctcttcgcag gcttcgccga cctgctcgcg
     4141 ccacttcttc acgcgggtgg aatccgatcc gcacatgagg cggaaggttt ccagcttgag
     4201 cgggtacggc tcccggtgcg agctgaaata gtcgaacatc cgtcgggccg tcggcgacag
     4261 cttgcggtac ttctcccata tgaatttcgt gtagtggtcg ccagcaaaca gcacgacgat
     4321 ttcctcgtcg atcaggacct ggcaacggga cgttttcttg ccacggtcca ggacgcggaa
     4381 gcggtgcagc agcgacaccg attccaggtg cccaacgcgg tcggacgtga agcccatcgc
     4441 cgtcgcctgt aggcgcgaca ggcattcctc ggccttcgtg taataccggc cattgatcga
     4501 ccagcccagg tcctggcaaa gctcgtagaa cgtgaaggtg atcggctcgc cgataggggt
     4561 gcgcttcgcg tactccaaca cctgctgcca caccagttcg tcatcgtcgg cccgcagctc
     4621 gacgccggtg taggtgatct tcacgtcctt gttgacgtgg aaaatgacct tgttttgcag
     4681 cgcctcgcgc gggattttct tgttgcgcgt ggtgaacagg gcagagcggg ccgtgtcgtt
     4741 tggcatcgct cgcatcgtgt ccggccacgg cgcaatatcg aacaaggaaa gctgcatttc
     4801 cttgatctgc tgcttcgtgt gtttcagcaa cgcggcctgc ttggcctcgc tgacctgttt
     4861 tgccaggtcc tcgccggcgg tttttcgctt cttggtcgtc atagttcctc gcgtgtcgat
     4921 ggtcatcgac ttcgccaaac ctgccgcctc ctgttcaaga cgacgcgaac gctccacggc
     4981 ggccgatggc gcgggcaggg cagggggagc cagttgcacg ctgtcgcgct cgatcttggc
     5041 cgtagcttgc tggaccatcg agccgacgga ctggaaggtt tcgcggggcg cacgcatgac
     5101 ggtgcggctt gcgatggttt cggcatcctc ggcggaaaac cccgcgtcga tcagttcttg
     5161 cctgtatgcc ttccggtcaa acgtccgatt cattcaccct ccttgcggga ttgccccgac
     5221 tcacgccggg gcaatgtgcc cttattcctg atttgacccg cctggtgcct tggtgtccag
     5281 ataatccacc ttatcggcaa tgaagtcggt cccgtagacc gtctggccgt ccttctcgta
     5341 cttggtattc cgaatcttgc cctgcacgaa taccagcgac cccttgccca aatacttgcc
     5401 gtgggcctcg gcctgagagc caaaacactt gatgcggaag aagtcggtgc gctcctgctt
     5461 gtcgccggca tcgttgcgcc acatctagga tctgccagga accgtaaaaa ggccgcgttg
     5521 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5581 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5641 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5701 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5761 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5821 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5881 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5941 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     6001 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6061 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6121 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6181 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6241 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6301 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6361 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6421 ataccgcgag aaccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6481 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6541 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6601 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6661 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6721 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6781 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6841 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6901 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6961 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7021 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7081 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7141 atactcatac tcttcctttt tcaatattat tgaagcattt atcaaggtta ttgtctcatg
     7201 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacgaat
     7261 tggccagcgc tgccattttt ggggtgaggc cgttcgcggc cgaggggcgc agcccctggg
     7321 gggatgggag gcccgcgtt
//
