LOCUS       sequence-318519-        7029 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 7029)
  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 7029)
  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: pEPYC1CB0437
FEATURES             Location/Qualifiers
     source          1..7029
                     /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    995..1050
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             1052..2701
                     /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(1118..1136)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     CDS             2765..2788
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      2799..3506
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_feature    3555..3579
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(3735..4883)
                     /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(5219..5807)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5299..5318)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5978..6769,6770..6838))
                     /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     6601..6620
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6839..6943)
                     /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 ttagtgaggt ggtggagcac gacatggtgg
      901 agcacgacat ggtggagcac gacactatat aaggttttgc tattcattga aagcagtagt
      961 gactgatttg tatataatgt actgtatttt tacaacaatt accaacaaca acaaacaaca
     1021 aacaacatta caattactat ttacaattac aatggaggac gccaaaaaca taaagaaagg
     1081 cccggcgcca ttctatccgc tggaagatgg aaccgctgga gagcaactgc ataaggctat
     1141 gaagagatac gccctggttc ctggaacaat tgcttttaca gatgcacata tcgaggtgga
     1201 catcacttac gctgagtact tcgaaatgtc cgttcggttg gcagaagcta tgaaacgata
     1261 tgggctgaat acaaatcaca gaatcgtcgt atgcagtgaa aactctcttc aattctttat
     1321 gccggtgttg ggcgcgttat ttatcggagt tgcagttgcg cccgcgaacg acatttataa
     1381 tgaacgtgaa ttgctcaaca gtatgggcat ttcgcagcct accgtggtgt tcgtttccaa
     1441 aaaggggttg caaaaaattt tgaacgtgca aaaaaagctc ccaatcatcc aaaaaattat
     1501 tatcatggat tctaaaacgg attaccaggg atttcagtcg atgtacacgt tcgtcacatc
     1561 tcatctacct cccggtttta atgaatacga ttttgtgcca gagtccttcg atagggacaa
     1621 gacaattgca ctgatcatga actcctctgg atctactggt ctgcctaaag gtgtcgctct
     1681 gcctcataga actgcctgcg tgagattctc gcatgccaga gatcctattt ttggcaatca
     1741 aatcattccg gatactgcga ttttaagtgt tgttccattc catcacggtt ttggaatgtt
     1801 tactacactc ggatatttga tatgtggatt tcgagtcgtc ttaatgtata gatttgaaga
     1861 agaactgttt ctgaggagcc ttcaggatta caagattcaa agtgcgctgc tggtgccaac
     1921 cctattctcc ttcttcgcca aaagcactct gattgacaaa tacgatttat ctaatttaca
     1981 cgaaattgct tctggtggcg ctcccctctc taaggaagtc ggggaagcgg ttgccaagag
     2041 gttccatctg ccaggtatca ggcaaggata tgggctcact gagactacat cagctattct
     2101 gattacaccc gagggggatg ataaaccggg cgcggtcggt aaagttgttc cattttttga
     2161 agcgaaggtt gtggatctgg ataccgggaa aacgctgggc gttaatcaaa gaggcgaact
     2221 gtgtgtgaga ggtcctatga ttatgtccgg ttatgtaaac aatccggaag cgaccaacgc
     2281 cttgattgac aaggatggat ggctacattc tggagacata gcttactggg acgaggacga
     2341 acacttcttc atcgttgacc gcctgaagtc tctgattaag tacaaaggct atcaggtggc
     2401 tcccgctgaa ttggaatcca tcttgctcca acaccccaac atcttcgacg caggtgtcgc
     2461 aggtctgccc gacgatgacg ccggtgaact tcccgccgcc gttgttgttt tggagcacgg
     2521 aaagacgatg acggaaaaag agatcgtgga ttacgtcgcc agtcaagtaa caaccgcgaa
     2581 aaagttgcgc ggaggagttg tgtttgtgga cgaagtaccg aaaggtctta ccggaaaact
     2641 cgacgcaaga aaaatcagag agatcctcat aaaggccaag aagggcggaa agatcgccgt
     2701 gggttcggat tataaggacc atgacggaga ctataaggac catgacctcg acgctgcagc
     2761 agcggattat aaggacgatg acgataagtg agcttgtcct gctttaatga gatatgcgag
     2821 aagcctatga tcgcatgata tttgctttca attctgttgt gcacgttgta aaaaacctga
     2881 gcatgtgtag ctcagatcct taccgccggt ttcggttcat tctaatgaat atatcacccg
     2941 ttactatcgt atttttatga ataatattct ccgttcaatt tactgattgt accctactac
     3001 ttatatgtac aatattaaaa tgaaaacaat atattgtgct gaataggttt atagcgacat
     3061 ctatgataga gcgccacaat aacaaacaat tgcgttttat tattacaaat ccaattttaa
     3121 aaaaagcggc agaaccggtc aaacctaaaa gactgattac ataaatctta ttcaaatttc
     3181 aaaagtgccc caggggctag tatctacgac acaccgagcg gcgaactaat aacgctcact
     3241 gaagggaact ccggttcccc gccggcgcgc atgggtgaga ttccttgaag ttgagtattg
     3301 gccgtccgct ctaccgaaag ttacgggcac cattcaaccc ggtccagcac ggcggccggg
     3361 taaccgactt gctgccccga gaattatgca gcattttttt ggtgtatgtg ggccccaaat
     3421 gaagtgcagg tcaaaccttg acagtgacga caaatcgttg ggcgggtcca gggcgaattt
     3481 tgcgacaaca tgtcgaggct cagcaggacc gctgcaattg tcttctgcac gaagtggttt
     3541 aaactatcag tgtttgacag gatatattgg cgggtaaacc taagagaaaa gagcgtttat
     3601 tagaataatc ggatatttaa aagggcgtga aaaggtttat ccgttcgtcc atttgtatgt
     3661 gcatgccaac cacagggttc cccagatcag gcgctggctg ctgaaccccc agccggaact
     3721 gaccccacaa ggccctagcg tttgcaatgc accaggtcat cattgaccca ggcgtgttcc
     3781 accaggccgc tgcctcgcaa ctcttcgcag gcttcgccga cctgctcgcg ccacttcttc
     3841 acgcgggtgg aatccgatcc gcacatgagg cggaaggttt ccagcttgag cgggtacggc
     3901 tcccggtgcg agctgaaata gtcgaacatc cgtcgggccg tcggcgacag cttgcggtac
     3961 ttctcccata tgaatttcgt gtagtggtcg ccagcaaaca gcacgacgat ttcctcgtcg
     4021 atcaggacct ggcaacggga cgttttcttg ccacggtcca ggacgcggaa gcggtgcagc
     4081 agcgacaccg attccaggtg cccaacgcgg tcggacgtga agcccatcgc cgtcgcctgt
     4141 aggcgcgaca ggcattcctc ggccttcgtg taataccggc cattgatcga ccagcccagg
     4201 tcctggcaaa gctcgtagaa cgtgaaggtg atcggctcgc cgataggggt gcgcttcgcg
     4261 tactccaaca cctgctgcca caccagttcg tcatcgtcgg cccgcagctc gacgccggtg
     4321 taggtgatct tcacgtcctt gttgacgtgg aaaatgacct tgttttgcag cgcctcgcgc
     4381 gggattttct tgttgcgcgt ggtgaacagg gcagagcggg ccgtgtcgtt tggcatcgct
     4441 cgcatcgtgt ccggccacgg cgcaatatcg aacaaggaaa gctgcatttc cttgatctgc
     4501 tgcttcgtgt gtttcagcaa cgcggcctgc ttggcctcgc tgacctgttt tgccaggtcc
     4561 tcgccggcgg tttttcgctt cttggtcgtc atagttcctc gcgtgtcgat ggtcatcgac
     4621 ttcgccaaac ctgccgcctc ctgttcaaga cgacgcgaac gctccacggc ggccgatggc
     4681 gcgggcaggg cagggggagc cagttgcacg ctgtcgcgct cgatcttggc cgtagcttgc
     4741 tggaccatcg agccgacgga ctggaaggtt tcgcggggcg cacgcatgac ggtgcggctt
     4801 gcgatggttt cggcatcctc ggcggaaaac cccgcgtcga tcagttcttg cctgtatgcc
     4861 ttccggtcaa acgtccgatt cattcaccct ccttgcggga ttgccccgac tcacgccggg
     4921 gcaatgtgcc cttattcctg atttgacccg cctggtgcct tggtgtccag ataatccacc
     4981 ttatcggcaa tgaagtcggt cccgtagacc gtctggccgt ccttctcgta cttggtattc
     5041 cgaatcttgc cctgcacgaa taccagcgac cccttgccca aatacttgcc gtgggcctcg
     5101 gcctgagagc caaaacactt gatgcggaag aagtcggtgc gctcctgctt gtcgccggca
     5161 tcgttgcgcc acatctagga tctgccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5221 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5281 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5341 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5401 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5461 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5521 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5581 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5641 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5701 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5761 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5821 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5881 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5941 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6001 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6061 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6121 aaccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6181 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6241 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6301 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6361 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6421 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6481 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6541 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6601 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6661 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6721 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6781 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6841 tcttcctttt tcaatattat tgaagcattt atcaaggtta ttgtctcatg agcggataca
     6901 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacgaat tggccagcgc
     6961 tgccattttt ggggtgaggc cgttcgcggc cgaggggcgc agcccctggg gggatgggag
     7021 gcccgcgtt
//
