LOCUS       sequence-330625-        7307 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 7307)
  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 7307)
  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: pEPYC1CB0173
FEATURES             Location/Qualifiers
     source          1..7307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..47
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     misc_feature    68..123
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             125..1774
                     /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(191..209)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     CDS             1838..1861
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      1872..2579
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_feature    2628..2652
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2808..3956)
                     /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(4292..4880)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4372..4391)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5051..5842,5843..5911))
                     /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     5674..5693
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5912..6016)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6103..6734
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    6826..6850
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     join(7299..7307,1..12)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
ORIGIN
        1 gcaagaccct tcctctatat aaggaagttc atttcatttg gagaggacac gctactgtat
       61 ttttacaaca attaccaaca acaacaaaca acaaacaaca ttacaattac tatttacaat
      121 tacaatggag gacgccaaaa acataaagaa aggcccggcg ccattctatc cgctggaaga
      181 tggaaccgct ggagagcaac tgcataaggc tatgaagaga tacgccctgg ttcctggaac
      241 aattgctttt acagatgcac atatcgaggt ggacatcact tacgctgagt acttcgaaat
      301 gtccgttcgg ttggcagaag ctatgaaacg atatgggctg aatacaaatc acagaatcgt
      361 cgtatgcagt gaaaactctc ttcaattctt tatgccggtg ttgggcgcgt tatttatcgg
      421 agttgcagtt gcgcccgcga acgacattta taatgaacgt gaattgctca acagtatggg
      481 catttcgcag cctaccgtgg tgttcgtttc caaaaagggg ttgcaaaaaa ttttgaacgt
      541 gcaaaaaaag ctcccaatca tccaaaaaat tattatcatg gattctaaaa cggattacca
      601 gggatttcag tcgatgtaca cgttcgtcac atctcatcta cctcccggtt ttaatgaata
      661 cgattttgtg ccagagtcct tcgataggga caagacaatt gcactgatca tgaactcctc
      721 tggatctact ggtctgccta aaggtgtcgc tctgcctcat agaactgcct gcgtgagatt
      781 ctcgcatgcc agagatccta tttttggcaa tcaaatcatt ccggatactg cgattttaag
      841 tgttgttcca ttccatcacg gttttggaat gtttactaca ctcggatatt tgatatgtgg
      901 atttcgagtc gtcttaatgt atagatttga agaagaactg tttctgagga gccttcagga
      961 ttacaagatt caaagtgcgc tgctggtgcc aaccctattc tccttcttcg ccaaaagcac
     1021 tctgattgac aaatacgatt tatctaattt acacgaaatt gcttctggtg gcgctcccct
     1081 ctctaaggaa gtcggggaag cggttgccaa gaggttccat ctgccaggta tcaggcaagg
     1141 atatgggctc actgagacta catcagctat tctgattaca cccgaggggg atgataaacc
     1201 gggcgcggtc ggtaaagttg ttccattttt tgaagcgaag gttgtggatc tggataccgg
     1261 gaaaacgctg ggcgttaatc aaagaggcga actgtgtgtg agaggtccta tgattatgtc
     1321 cggttatgta aacaatccgg aagcgaccaa cgccttgatt gacaaggatg gatggctaca
     1381 ttctggagac atagcttact gggacgagga cgaacacttc ttcatcgttg accgcctgaa
     1441 gtctctgatt aagtacaaag gctatcaggt ggctcccgct gaattggaat ccatcttgct
     1501 ccaacacccc aacatcttcg acgcaggtgt cgcaggtctg cccgacgatg acgccggtga
     1561 acttcccgcc gccgttgttg ttttggagca cggaaagacg atgacggaaa aagagatcgt
     1621 ggattacgtc gccagtcaag taacaaccgc gaaaaagttg cgcggaggag ttgtgtttgt
     1681 ggacgaagta ccgaaaggtc ttaccggaaa actcgacgca agaaaaatca gagagatcct
     1741 cataaaggcc aagaagggcg gaaagatcgc cgtgggttcg gattataagg accatgacgg
     1801 agactataag gaccatgacc tcgacgctgc agcagcggat tataaggacg atgacgataa
     1861 gtgagcttgt cctgctttaa tgagatatgc gagaagccta tgatcgcatg atatttgctt
     1921 tcaattctgt tgtgcacgtt gtaaaaaacc tgagcatgtg tagctcagat ccttaccgcc
     1981 ggtttcggtt cattctaatg aatatatcac ccgttactat cgtattttta tgaataatat
     2041 tctccgttca atttactgat tgtaccctac tacttatatg tacaatatta aaatgaaaac
     2101 aatatattgt gctgaatagg tttatagcga catctatgat agagcgccac aataacaaac
     2161 aattgcgttt tattattaca aatccaattt taaaaaaagc ggcagaaccg gtcaaaccta
     2221 aaagactgat tacataaatc ttattcaaat ttcaaaagtg ccccaggggc tagtatctac
     2281 gacacaccga gcggcgaact aataacgctc actgaaggga actccggttc cccgccggcg
     2341 cgcatgggtg agattccttg aagttgagta ttggccgtcc gctctaccga aagttacggg
     2401 caccattcaa cccggtccag cacggcggcc gggtaaccga cttgctgccc cgagaattat
     2461 gcagcatttt tttggtgtat gtgggcccca aatgaagtgc aggtcaaacc ttgacagtga
     2521 cgacaaatcg ttgggcgggt ccagggcgaa ttttgcgaca acatgtcgag gctcagcagg
     2581 accgctgcaa ttgtcttctg cacgaagtgg tttaaactat cagtgtttga caggatatat
     2641 tggcgggtaa acctaagaga aaagagcgtt tattagaata atcggatatt taaaagggcg
     2701 tgaaaaggtt tatccgttcg tccatttgta tgtgcatgcc aaccacaggg ttccccagat
     2761 caggcgctgg ctgctgaacc cccagccgga actgacccca caaggcccta gcgtttgcaa
     2821 tgcaccaggt catcattgac ccaggcgtgt tccaccaggc cgctgcctcg caactcttcg
     2881 caggcttcgc cgacctgctc gcgccacttc ttcacgcggg tggaatccga tccgcacatg
     2941 aggcggaagg tttccagctt gagcgggtac ggctcccggt gcgagctgaa atagtcgaac
     3001 atccgtcggg ccgtcggcga cagcttgcgg tacttctccc atatgaattt cgtgtagtgg
     3061 tcgccagcaa acagcacgac gatttcctcg tcgatcagga cctggcaacg ggacgttttc
     3121 ttgccacggt ccaggacgcg gaagcggtgc agcagcgaca ccgattccag gtgcccaacg
     3181 cggtcggacg tgaagcccat cgccgtcgcc tgtaggcgcg acaggcattc ctcggccttc
     3241 gtgtaatacc ggccattgat cgaccagccc aggtcctggc aaagctcgta gaacgtgaag
     3301 gtgatcggct cgccgatagg ggtgcgcttc gcgtactcca acacctgctg ccacaccagt
     3361 tcgtcatcgt cggcccgcag ctcgacgccg gtgtaggtga tcttcacgtc cttgttgacg
     3421 tggaaaatga ccttgttttg cagcgcctcg cgcgggattt tcttgttgcg cgtggtgaac
     3481 agggcagagc gggccgtgtc gtttggcatc gctcgcatcg tgtccggcca cggcgcaata
     3541 tcgaacaagg aaagctgcat ttccttgatc tgctgcttcg tgtgtttcag caacgcggcc
     3601 tgcttggcct cgctgacctg ttttgccagg tcctcgccgg cggtttttcg cttcttggtc
     3661 gtcatagttc ctcgcgtgtc gatggtcatc gacttcgcca aacctgccgc ctcctgttca
     3721 agacgacgcg aacgctccac ggcggccgat ggcgcgggca gggcaggggg agccagttgc
     3781 acgctgtcgc gctcgatctt ggccgtagct tgctggacca tcgagccgac ggactggaag
     3841 gtttcgcggg gcgcacgcat gacggtgcgg cttgcgatgg tttcggcatc ctcggcggaa
     3901 aaccccgcgt cgatcagttc ttgcctgtat gccttccggt caaacgtccg attcattcac
     3961 cctccttgcg ggattgcccc gactcacgcc ggggcaatgt gcccttattc ctgatttgac
     4021 ccgcctggtg ccttggtgtc cagataatcc accttatcgg caatgaagtc ggtcccgtag
     4081 accgtctggc cgtccttctc gtacttggta ttccgaatct tgccctgcac gaataccagc
     4141 gaccccttgc ccaaatactt gccgtgggcc tcggcctgag agccaaaaca cttgatgcgg
     4201 aagaagtcgg tgcgctcctg cttgtcgccg gcatcgttgc gccacatcta ggatctgcca
     4261 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4321 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4381 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4441 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4501 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4561 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4621 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4681 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4741 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4801 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4861 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4921 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4981 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5041 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5101 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5161 catctggccc cagtgctgca atgataccgc gagaaccacg ctcaccggct ccagatttat
     5221 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5281 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5341 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5401 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5461 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5521 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5581 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5641 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5701 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5761 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5821 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5881 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     5941 tttatcaagg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6001 aaataggggt tccgcgcacg aattggccag cgctgccatt tttggggtga ggccgttcgc
     6061 ggccgagggg cgcagcccct ggggggatgg gaggcccgcg ttagcgggcc gggagggttc
     6121 gagaaggggg ggcacccccc ttcggcgtgc gcggtcacgc gcacagggcg cagccctggt
     6181 taaaaacaag gtttataaat attggtttaa aagcaggtta aaagacaggt tagcggtggc
     6241 cgaaaaacgg gcggaaaccc ttgcaaatgc tggattttct gcctgtggac agcccctcaa
     6301 atgtcaatag gtgcgcccct catctgtcag cactctgccc ctcaagtgtc aaggatcgcg
     6361 cccctcatct gtcagtagtc gcgcccctca agtgtcaata ccgcagggca cttatcccca
     6421 ggcttgtcca catcatctgt gggaaactcg cgtaaaatca ggcgttttcg ccgatttgcg
     6481 aggctggcca gctccacgtc gccggccgaa atcgagcctg cccctcatct gtcaacgccg
     6541 cgccgggtga gtcggcccct caagtgtcaa cgtccgcccc tcatctgtca gtgagggcca
     6601 agttttccgc gaggtatcca caacgccggc ggccgcggtg tctcgcacac ggcttcgacg
     6661 gcgtttctgg cgcgtttgca gggccataga cggccgccag cccagcggcg agggcaacca
     6721 gcccggtgag cgtcgcaaag gagatcctga tctgactgat gggctgcctg tatcgagtgg
     6781 tgattttgtg ccgagctgcc ggtcggggag ctgttggctg gctggtggca ggatatattg
     6841 tggtgtaaac aaattgacgc ttagacaact taataacaca ttgcggacgt ttttaatgta
     6901 ctggggtgga tgcagtgggc cccactctgt gaagacaatg ccgaattcgg atccggaggt
     6961 caacatggtg gagcacgaca ctctggtcta ctccaaaaat gtcaaagata cagtctcaga
     7021 agatcaaagg gctattgaga cttttcaaca aaggataatt tcgggaaacc tcctcggatt
     7081 ccattgccca gctatctgtc acttcatcga aaggacagta gaaaaggaag gtggctccta
     7141 caaatgccat cattgcgata aaggaaaggc tatcattcaa gagatggacc cccacccacg
     7201 aggagcatcg tggaaaaaga agaggttcca accacgtcta caaagcaagt ggattgatgt
     7261 gacatctcca ctgacgtaag ggatgacgca caatcccact atccttc
//
