LOCUS       sequence-318551-        7027 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 7027)
  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 7027)
  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: pEPYC1CB0514
FEATURES             Location/Qualifiers
     source          1..7027
                     /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    993..1048
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             1050..2699
                     /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(1116..1134)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     CDS             2763..2786
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      2797..3504
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_feature    3553..3577
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(3733..4881)
                     /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(5217..5805)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5297..5316)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5976..6767,6768..6836))
                     /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     6599..6618
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6837..6941)
                     /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 ttagtgagga accattattg cgatgagcta
      901 agcacatacg tcagtcacta tcagcctata taaggttttg ctattcattg aaagcagtag
      961 tgactgattt gtatatatac tgtattttta caacaattac caacaacaac aaacaacaaa
     1021 caacattaca attactattt acaattacaa tggaggacgc caaaaacata aagaaaggcc
     1081 cggcgccatt ctatccgctg gaagatggaa ccgctggaga gcaactgcat aaggctatga
     1141 agagatacgc cctggttcct ggaacaattg cttttacaga tgcacatatc gaggtggaca
     1201 tcacttacgc tgagtacttc gaaatgtccg ttcggttggc agaagctatg aaacgatatg
     1261 ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa ttctttatgc
     1321 cggtgttggg cgcgttattt atcggagttg cagttgcgcc cgcgaacgac atttataatg
     1381 aacgtgaatt gctcaacagt atgggcattt cgcagcctac cgtggtgttc gtttccaaaa
     1441 aggggttgca aaaaattttg aacgtgcaaa aaaagctccc aatcatccaa aaaattatta
     1501 tcatggattc taaaacggat taccagggat ttcagtcgat gtacacgttc gtcacatctc
     1561 atctacctcc cggttttaat gaatacgatt ttgtgccaga gtccttcgat agggacaaga
     1621 caattgcact gatcatgaac tcctctggat ctactggtct gcctaaaggt gtcgctctgc
     1681 ctcatagaac tgcctgcgtg agattctcgc atgccagaga tcctattttt ggcaatcaaa
     1741 tcattccgga tactgcgatt ttaagtgttg ttccattcca tcacggtttt ggaatgttta
     1801 ctacactcgg atatttgata tgtggatttc gagtcgtctt aatgtataga tttgaagaag
     1861 aactgtttct gaggagcctt caggattaca agattcaaag tgcgctgctg gtgccaaccc
     1921 tattctcctt cttcgccaaa agcactctga ttgacaaata cgatttatct aatttacacg
     1981 aaattgcttc tggtggcgct cccctctcta aggaagtcgg ggaagcggtt gccaagaggt
     2041 tccatctgcc aggtatcagg caaggatatg ggctcactga gactacatca gctattctga
     2101 ttacacccga gggggatgat aaaccgggcg cggtcggtaa agttgttcca ttttttgaag
     2161 cgaaggttgt ggatctggat accgggaaaa cgctgggcgt taatcaaaga ggcgaactgt
     2221 gtgtgagagg tcctatgatt atgtccggtt atgtaaacaa tccggaagcg accaacgcct
     2281 tgattgacaa ggatggatgg ctacattctg gagacatagc ttactgggac gaggacgaac
     2341 acttcttcat cgttgaccgc ctgaagtctc tgattaagta caaaggctat caggtggctc
     2401 ccgctgaatt ggaatccatc ttgctccaac accccaacat cttcgacgca ggtgtcgcag
     2461 gtctgcccga cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg gagcacggaa
     2521 agacgatgac ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca accgcgaaaa
     2581 agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa aggtcttacc ggaaaactcg
     2641 acgcaagaaa aatcagagag atcctcataa aggccaagaa gggcggaaag atcgccgtgg
     2701 gttcggatta taaggaccat gacggagact ataaggacca tgacctcgac gctgcagcag
     2761 cggattataa ggacgatgac gataagtgag cttgtcctgc tttaatgaga tatgcgagaa
     2821 gcctatgatc gcatgatatt tgctttcaat tctgttgtgc acgttgtaaa aaacctgagc
     2881 atgtgtagct cagatcctta ccgccggttt cggttcattc taatgaatat atcacccgtt
     2941 actatcgtat ttttatgaat aatattctcc gttcaattta ctgattgtac cctactactt
     3001 atatgtacaa tattaaaatg aaaacaatat attgtgctga ataggtttat agcgacatct
     3061 atgatagagc gccacaataa caaacaattg cgttttatta ttacaaatcc aattttaaaa
     3121 aaagcggcag aaccggtcaa acctaaaaga ctgattacat aaatcttatt caaatttcaa
     3181 aagtgcccca ggggctagta tctacgacac accgagcggc gaactaataa cgctcactga
     3241 agggaactcc ggttccccgc cggcgcgcat gggtgagatt ccttgaagtt gagtattggc
     3301 cgtccgctct accgaaagtt acgggcacca ttcaacccgg tccagcacgg cggccgggta
     3361 accgacttgc tgccccgaga attatgcagc atttttttgg tgtatgtggg ccccaaatga
     3421 agtgcaggtc aaaccttgac agtgacgaca aatcgttggg cgggtccagg gcgaattttg
     3481 cgacaacatg tcgaggctca gcaggaccgc tgcaattgtc ttctgcacga agtggtttaa
     3541 actatcagtg tttgacagga tatattggcg ggtaaaccta agagaaaaga gcgtttatta
     3601 gaataatcgg atatttaaaa gggcgtgaaa aggtttatcc gttcgtccat ttgtatgtgc
     3661 atgccaacca cagggttccc cagatcaggc gctggctgct gaacccccag ccggaactga
     3721 ccccacaagg ccctagcgtt tgcaatgcac caggtcatca ttgacccagg cgtgttccac
     3781 caggccgctg cctcgcaact cttcgcaggc ttcgccgacc tgctcgcgcc acttcttcac
     3841 gcgggtggaa tccgatccgc acatgaggcg gaaggtttcc agcttgagcg ggtacggctc
     3901 ccggtgcgag ctgaaatagt cgaacatccg tcgggccgtc ggcgacagct tgcggtactt
     3961 ctcccatatg aatttcgtgt agtggtcgcc agcaaacagc acgacgattt cctcgtcgat
     4021 caggacctgg caacgggacg ttttcttgcc acggtccagg acgcggaagc ggtgcagcag
     4081 cgacaccgat tccaggtgcc caacgcggtc ggacgtgaag cccatcgccg tcgcctgtag
     4141 gcgcgacagg cattcctcgg ccttcgtgta ataccggcca ttgatcgacc agcccaggtc
     4201 ctggcaaagc tcgtagaacg tgaaggtgat cggctcgccg ataggggtgc gcttcgcgta
     4261 ctccaacacc tgctgccaca ccagttcgtc atcgtcggcc cgcagctcga cgccggtgta
     4321 ggtgatcttc acgtccttgt tgacgtggaa aatgaccttg ttttgcagcg cctcgcgcgg
     4381 gattttcttg ttgcgcgtgg tgaacagggc agagcgggcc gtgtcgtttg gcatcgctcg
     4441 catcgtgtcc ggccacggcg caatatcgaa caaggaaagc tgcatttcct tgatctgctg
     4501 cttcgtgtgt ttcagcaacg cggcctgctt ggcctcgctg acctgttttg ccaggtcctc
     4561 gccggcggtt tttcgcttct tggtcgtcat agttcctcgc gtgtcgatgg tcatcgactt
     4621 cgccaaacct gccgcctcct gttcaagacg acgcgaacgc tccacggcgg ccgatggcgc
     4681 gggcagggca gggggagcca gttgcacgct gtcgcgctcg atcttggccg tagcttgctg
     4741 gaccatcgag ccgacggact ggaaggtttc gcggggcgca cgcatgacgg tgcggcttgc
     4801 gatggtttcg gcatcctcgg cggaaaaccc cgcgtcgatc agttcttgcc tgtatgcctt
     4861 ccggtcaaac gtccgattca ttcaccctcc ttgcgggatt gccccgactc acgccggggc
     4921 aatgtgccct tattcctgat ttgacccgcc tggtgccttg gtgtccagat aatccacctt
     4981 atcggcaatg aagtcggtcc cgtagaccgt ctggccgtcc ttctcgtact tggtattccg
     5041 aatcttgccc tgcacgaata ccagcgaccc cttgcccaaa tacttgccgt gggcctcggc
     5101 ctgagagcca aaacacttga tgcggaagaa gtcggtgcgc tcctgcttgt cgccggcatc
     5161 gttgcgccac atctaggatc tgccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5221 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5281 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5341 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5401 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5461 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5521 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5581 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5641 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5701 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5761 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5821 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5881 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5941 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6001 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6061 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagaa
     6121 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6181 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6241 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6301 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6361 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6421 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6481 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6541 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6601 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6661 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6721 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6781 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6841 ttcctttttc aatattattg aagcatttat caaggttatt gtctcatgag cggatacata
     6901 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacgaattg gccagcgctg
     6961 ccatttttgg ggtgaggccg ttcgcggccg aggggcgcag cccctggggg gatgggaggc
     7021 ccgcgtt
//
