LOCUS       sequence-342375-        7218 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 7218)
  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 7218)
  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: pEPYC1CB0199
FEATURES             Location/Qualifiers
     source          1..7218
                     /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        136..516
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     CDS             518..2167
                     /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(584..602)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     CDS             2231..2254
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      2265..2972
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_feature    3021..3045
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(3201..4349)
                     /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(4685..5273)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4765..4784)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5444..6235,6236..6304))
                     /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     6067..6086
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6305..6409)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6496..7127
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 tggcaggata tattgtggtg taaacaaatt gacgcttaga caacttaata acacattgcg
       61 gacgttttta atgtactggg gtggatgcag tgggccccac tctgtgaaga caatgccgaa
      121 ttcggatccg gagatttttc aaatcagtgc gcaagacgtg acgtaagtat ccgagtcagt
      181 ttttattttt ctactaattt ggtcgtttat ttcggcgtgt aggacatggc aaccgggcct
      241 gaatttcgcg ggtattctgt ttctattcca actttttctt gatccgcagc cattaacgac
      301 ttttgaatag atacgctgac acgccaagcc tcgctagtca aaagtgtacc aaacaacgct
      361 ttacagcaag aacggaatgc gcgtgacgct cgcggtgacg ccatttcgcc ttttcagaaa
      421 tggataaata gccttgcttc ctattatatc ttcccaaatt accaatacat tacactagca
      481 tctgaatttc ataaccaatc tcgatacacc aaatcgaatg gaggacgcca aaaacataaa
      541 gaaaggcccg gcgccattct atccgctgga agatggaacc gctggagagc aactgcataa
      601 ggctatgaag agatacgccc tggttcctgg aacaattgct tttacagatg cacatatcga
      661 ggtggacatc acttacgctg agtacttcga aatgtccgtt cggttggcag aagctatgaa
      721 acgatatggg ctgaatacaa atcacagaat cgtcgtatgc agtgaaaact ctcttcaatt
      781 ctttatgccg gtgttgggcg cgttatttat cggagttgca gttgcgcccg cgaacgacat
      841 ttataatgaa cgtgaattgc tcaacagtat gggcatttcg cagcctaccg tggtgttcgt
      901 ttccaaaaag gggttgcaaa aaattttgaa cgtgcaaaaa aagctcccaa tcatccaaaa
      961 aattattatc atggattcta aaacggatta ccagggattt cagtcgatgt acacgttcgt
     1021 cacatctcat ctacctcccg gttttaatga atacgatttt gtgccagagt ccttcgatag
     1081 ggacaagaca attgcactga tcatgaactc ctctggatct actggtctgc ctaaaggtgt
     1141 cgctctgcct catagaactg cctgcgtgag attctcgcat gccagagatc ctatttttgg
     1201 caatcaaatc attccggata ctgcgatttt aagtgttgtt ccattccatc acggttttgg
     1261 aatgtttact acactcggat atttgatatg tggatttcga gtcgtcttaa tgtatagatt
     1321 tgaagaagaa ctgtttctga ggagccttca ggattacaag attcaaagtg cgctgctggt
     1381 gccaacccta ttctccttct tcgccaaaag cactctgatt gacaaatacg atttatctaa
     1441 tttacacgaa attgcttctg gtggcgctcc cctctctaag gaagtcgggg aagcggttgc
     1501 caagaggttc catctgccag gtatcaggca aggatatggg ctcactgaga ctacatcagc
     1561 tattctgatt acacccgagg gggatgataa accgggcgcg gtcggtaaag ttgttccatt
     1621 ttttgaagcg aaggttgtgg atctggatac cgggaaaacg ctgggcgtta atcaaagagg
     1681 cgaactgtgt gtgagaggtc ctatgattat gtccggttat gtaaacaatc cggaagcgac
     1741 caacgccttg attgacaagg atggatggct acattctgga gacatagctt actgggacga
     1801 ggacgaacac ttcttcatcg ttgaccgcct gaagtctctg attaagtaca aaggctatca
     1861 ggtggctccc gctgaattgg aatccatctt gctccaacac cccaacatct tcgacgcagg
     1921 tgtcgcaggt ctgcccgacg atgacgccgg tgaacttccc gccgccgttg ttgttttgga
     1981 gcacggaaag acgatgacgg aaaaagagat cgtggattac gtcgccagtc aagtaacaac
     2041 cgcgaaaaag ttgcgcggag gagttgtgtt tgtggacgaa gtaccgaaag gtcttaccgg
     2101 aaaactcgac gcaagaaaaa tcagagagat cctcataaag gccaagaagg gcggaaagat
     2161 cgccgtgggt tcggattata aggaccatga cggagactat aaggaccatg acctcgacgc
     2221 tgcagcagcg gattataagg acgatgacga taagtgagct tgtcctgctt taatgagata
     2281 tgcgagaagc ctatgatcgc atgatatttg ctttcaattc tgttgtgcac gttgtaaaaa
     2341 acctgagcat gtgtagctca gatccttacc gccggtttcg gttcattcta atgaatatat
     2401 cacccgttac tatcgtattt ttatgaataa tattctccgt tcaatttact gattgtaccc
     2461 tactacttat atgtacaata ttaaaatgaa aacaatatat tgtgctgaat aggtttatag
     2521 cgacatctat gatagagcgc cacaataaca aacaattgcg ttttattatt acaaatccaa
     2581 ttttaaaaaa agcggcagaa ccggtcaaac ctaaaagact gattacataa atcttattca
     2641 aatttcaaaa gtgccccagg ggctagtatc tacgacacac cgagcggcga actaataacg
     2701 ctcactgaag ggaactccgg ttccccgccg gcgcgcatgg gtgagattcc ttgaagttga
     2761 gtattggccg tccgctctac cgaaagttac gggcaccatt caacccggtc cagcacggcg
     2821 gccgggtaac cgacttgctg ccccgagaat tatgcagcat ttttttggtg tatgtgggcc
     2881 ccaaatgaag tgcaggtcaa accttgacag tgacgacaaa tcgttgggcg ggtccagggc
     2941 gaattttgcg acaacatgtc gaggctcagc aggaccgctg caattgtctt ctgcacgaag
     3001 tggtttaaac tatcagtgtt tgacaggata tattggcggg taaacctaag agaaaagagc
     3061 gtttattaga ataatcggat atttaaaagg gcgtgaaaag gtttatccgt tcgtccattt
     3121 gtatgtgcat gccaaccaca gggttcccca gatcaggcgc tggctgctga acccccagcc
     3181 ggaactgacc ccacaaggcc ctagcgtttg caatgcacca ggtcatcatt gacccaggcg
     3241 tgttccacca ggccgctgcc tcgcaactct tcgcaggctt cgccgacctg ctcgcgccac
     3301 ttcttcacgc gggtggaatc cgatccgcac atgaggcgga aggtttccag cttgagcggg
     3361 tacggctccc ggtgcgagct gaaatagtcg aacatccgtc gggccgtcgg cgacagcttg
     3421 cggtacttct cccatatgaa tttcgtgtag tggtcgccag caaacagcac gacgatttcc
     3481 tcgtcgatca ggacctggca acgggacgtt ttcttgccac ggtccaggac gcggaagcgg
     3541 tgcagcagcg acaccgattc caggtgccca acgcggtcgg acgtgaagcc catcgccgtc
     3601 gcctgtaggc gcgacaggca ttcctcggcc ttcgtgtaat accggccatt gatcgaccag
     3661 cccaggtcct ggcaaagctc gtagaacgtg aaggtgatcg gctcgccgat aggggtgcgc
     3721 ttcgcgtact ccaacacctg ctgccacacc agttcgtcat cgtcggcccg cagctcgacg
     3781 ccggtgtagg tgatcttcac gtccttgttg acgtggaaaa tgaccttgtt ttgcagcgcc
     3841 tcgcgcggga ttttcttgtt gcgcgtggtg aacagggcag agcgggccgt gtcgtttggc
     3901 atcgctcgca tcgtgtccgg ccacggcgca atatcgaaca aggaaagctg catttccttg
     3961 atctgctgct tcgtgtgttt cagcaacgcg gcctgcttgg cctcgctgac ctgttttgcc
     4021 aggtcctcgc cggcggtttt tcgcttcttg gtcgtcatag ttcctcgcgt gtcgatggtc
     4081 atcgacttcg ccaaacctgc cgcctcctgt tcaagacgac gcgaacgctc cacggcggcc
     4141 gatggcgcgg gcagggcagg gggagccagt tgcacgctgt cgcgctcgat cttggccgta
     4201 gcttgctgga ccatcgagcc gacggactgg aaggtttcgc ggggcgcacg catgacggtg
     4261 cggcttgcga tggtttcggc atcctcggcg gaaaaccccg cgtcgatcag ttcttgcctg
     4321 tatgccttcc ggtcaaacgt ccgattcatt caccctcctt gcgggattgc cccgactcac
     4381 gccggggcaa tgtgccctta ttcctgattt gacccgcctg gtgccttggt gtccagataa
     4441 tccaccttat cggcaatgaa gtcggtcccg tagaccgtct ggccgtcctt ctcgtacttg
     4501 gtattccgaa tcttgccctg cacgaatacc agcgacccct tgcccaaata cttgccgtgg
     4561 gcctcggcct gagagccaaa acacttgatg cggaagaagt cggtgcgctc ctgcttgtcg
     4621 ccggcatcgt tgcgccacat ctaggatctg ccaggaaccg taaaaaggcc gcgttgctgg
     4681 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4741 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4801 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4861 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4921 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4981 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5041 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5101 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5161 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5221 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5281 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5341 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5401 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5461 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5521 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5581 cgcgagaacc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5641 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5701 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5761 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5821 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5881 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5941 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6001 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6061 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6121 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6181 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6241 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6301 tcatactctt cctttttcaa tattattgaa gcatttatca aggttattgt ctcatgagcg
     6361 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acgaattggc
     6421 cagcgctgcc atttttgggg tgaggccgtt cgcggccgag gggcgcagcc cctgggggga
     6481 tgggaggccc gcgttagcgg gccgggaggg ttcgagaagg gggggcaccc cccttcggcg
     6541 tgcgcggtca cgcgcacagg gcgcagccct ggttaaaaac aaggtttata aatattggtt
     6601 taaaagcagg ttaaaagaca ggttagcggt ggccgaaaaa cgggcggaaa cccttgcaaa
     6661 tgctggattt tctgcctgtg gacagcccct caaatgtcaa taggtgcgcc cctcatctgt
     6721 cagcactctg cccctcaagt gtcaaggatc gcgcccctca tctgtcagta gtcgcgcccc
     6781 tcaagtgtca ataccgcagg gcacttatcc ccaggcttgt ccacatcatc tgtgggaaac
     6841 tcgcgtaaaa tcaggcgttt tcgccgattt gcgaggctgg ccagctccac gtcgccggcc
     6901 gaaatcgagc ctgcccctca tctgtcaacg ccgcgccggg tgagtcggcc cctcaagtgt
     6961 caacgtccgc ccctcatctg tcagtgaggg ccaagttttc cgcgaggtat ccacaacgcc
     7021 ggcggccgcg gtgtctcgca cacggcttcg acggcgtttc tggcgcgttt gcagggccat
     7081 agacggccgc cagcccagcg gcgagggcaa ccagcccggt gagcgtcgca aaggagatcc
     7141 tgatctgact gatgggctgc ctgtatcgag tggtgatttt gtgccgagct gccggtcggg
     7201 gagctgttgg ctggctgg
//
