LOCUS       sequence-299497-        7676 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid level alpha luciferase reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7676)
  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 7676)
  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: pEPKK-alpha-2KN0102
FEATURES             Location/Qualifiers
     source          1..7676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    48..72
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     protein_bind    107..140
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    166..199
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    225..258
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    284..317
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    347..380
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    406..439
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        456..502
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             578..1090
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             1151..1174
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      1197..1397
                     /label=gene 7 terminator
                     /note="transcription terminator for octopine-type Ti 
                     plasmid gene 7"
     misc_feature    1415..1439
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             2740..3369
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIGTHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3803..4870
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      4936..5130
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(5366..5388)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5474..5614
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5529..5548
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5629..5646)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5800..6388)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5880..5899)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6476..7291)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(6476..7291)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     7199..7218
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 ggtgattttg tgccgagctg ccggtcgggg agctgttggc tggctggtgg caggatatat
       61 tgtggtgtaa acataacaag cttcgtctca gtcaggagcg gtgcgggtgc cagggcgtgc
      121 ccttgggctc cccgggcgcg tactccactc cccgagccgg tgcgggtgcc agggcgtgcc
      181 cttgggctcc ccgggcgcgt actccactcc cctgaccggt gcgggtgcca gggcgtgccc
      241 ttgggctccc cgggcgcgta ctccactccc cgagccggtg cgggtgccag ggcgtgccct
      301 tgggctcccc gggcgcgtac tccactcccc ccatggtgcg gtgcgggtgc cagggcgtgc
      361 ccttgggctc cccgggcgcg tactccactc cccgagccgg tgcgggtgcc agggcgtgcc
      421 cttgggctcc ccgggcgcgt actccactcc ccttcgcaag acccttcctc tatataagga
      481 agttcatttc atttggagag gactccggta tttttacaac aattaccaca acaaaacaaa
      541 caacaaacaa cattacaatt tactattcta gtcgaaaatg gtattcacac tcgaagattt
      601 cgttggggac tggcgacaga cagccggcta caacctggac caagtccttg aacagggagg
      661 tgtgtccagt ttgtttcaga atctcggggt gtccgtaact ccgatccaaa ggattgtcct
      721 gagcggtgaa aatgggctga agatcgacat ccatgtcatc atcccgtatg aaggtctgag
      781 cggcgaccaa atgggccaga tcgaaaaaat ttttaaggtg gtgtaccctg tggatgatca
      841 tcactttaag gtgatcctgc actatggcac actggtaatc gacggggtta cgccgaacat
      901 gatcgactat ttcggacggc cgtatgaagg catcgccgtg ttcgacggca aaaagatcac
      961 tgtaacaggg accctgtgga acggcaacaa aattatcgac gagcgcctga tcaaccccga
     1021 cggctccctg ctgttccgag taaccatcaa cggagtgacc ggctggcggc tgtgcgaacg
     1081 cattctggct tcggattata aggaccatga cggagactat aaggaccatg acctcgacgc
     1141 tgcagcagcg gattataagg acgatgacga taagtgagct tgatcccccg tcgacagcta
     1201 gctatatcat caatttatgt attacacata atatcgcact cagtctttca tctacggcaa
     1261 tgtaccagct gatataatca gttattgaaa tatttctgaa tttaaacttg catcaataaa
     1321 tttatgtttt tgcttggact ataatacctg acttgttatt ttatcaataa atatttaaac
     1381 tatatttctt tcaagatcgc ttgagacgaa gctttgacag gatatattgg cgggtaaacc
     1441 taagagaaaa gagcgtttat tagaataatc ggatatttaa aagggcgtga aaaggtttat
     1501 ccgttcgtcc atttgtatgt gcatgccaac cacagggttc ccctcgggat caaagtactt
     1561 tgatccaacc cctccgctgc tatagtgcag tcggcttctg acgttcagtg cagccgtcat
     1621 ctgaaaacga catgtcgcac aagtcctaag ttacgcgaca ggctgccgcc ctgccctttt
     1681 cctggcgttt tcttgtcgcg tgttttagtc gcataaagta gaatacttgc gactagaacc
     1741 ggagacatta cgccatgaac aagagcgccg ccgctggcct gctgggctat gcccgcgtca
     1801 gcaccgacga ccaggacttg accaaccaac gggccgaact gcacgcggcc ggctgcacca
     1861 agctgttttc cgagaagatc accggcacca ggcgcgaccg cccggagctg gccaggatgc
     1921 ttgaccacct acgccctggc gacgttgtga cagtgaccag gctagaccgc ctggcccgca
     1981 gcacccgcga cctactggac attgccgagc gcatccagga ggccggcgcg ggcctgcgta
     2041 gcctggcaga gccgtgggcc gacaccacca cgccggccgg ccgcatggtg ttgaccgtgt
     2101 tcgccggcat tgccgagttc gagcgttccc taatcatcga ccgcacccgg agcgggcgcg
     2161 aggccgccaa ggcccgaggc gtgaagtttg gcccccgccc taccctcacc ccggcacaga
     2221 tcgcgcacgc ccgcgagctg atcgaccagg aaggccgcac cgtgaaagag gcggctgcac
     2281 tgcttggcgt gcatcgctcg accctgtacc gcgcacttga gcgcagcgag gaagtgacgc
     2341 ccaccgaggc caggcggcgc ggtgccttcc gtgaggacgc attgaccgag gccgacgccc
     2401 tggcggccgc cgagaatgaa cgccaagagg aacaagcatg aaaccgcacc aggacggcca
     2461 ggacgaaccg tttttcatta ccgaagagat cgaggcggag atgatcgcgg ccgggtacgt
     2521 gttcgagccg cccgcgcacc tctcaaccgt gcggctgcat gaaatcctgg ccggtttgtc
     2581 tgatgccaag ctggcggcct ggccggccag cttggccgct gaagaaaccg agcgccgccg
     2641 tctaaaaagg tgatgtgtat ttgagtaaaa cagcttgcgt catgcggtcg ctgcgtatat
     2701 gatccgatga gtaaataaac aaatacgcaa ggggaacgca tgaaggttat cgctgtactt
     2761 aaccagaaag gcgggtcagg caagacgacc atcggaaccc atctagcccg cgccctgcaa
     2821 ctcgccgggg ccgatgttct gttagtcgat tccgatcccc agggcagtgc ccgcgattgg
     2881 gcggccgtgc gggaagatca accgctaacc gttgtcggca tcgaccgccc gacgattgac
     2941 cgcgacgtga aggccatcgg ccggcgcgac ttcgtagtga tcgacggagc gccccaggcg
     3001 gcggacttgg ctgtgtccgc gatcaaggca gccgacttcg tgctgattcc ggtgcagcca
     3061 agcccttacg acatatgggc caccgccgac ctggtggagc tggttaagca gcgcattgag
     3121 gtcacggatg gaaggctaca agcggccttt gtcgtgtcgc gggcgatcaa aggcacgcgc
     3181 atcggcggtg aggttgccga ggcgctggcc gggtacgagc tgcccattct tgagtcccgt
     3241 atcacgcagc gcgtgagcta cccaggcact gccgccgccg gcacaaccgt tcttgaatca
     3301 gaacccgagg gcgacgctgc ccgcgaggtc caggcgctgg ccgctgaaat taaatcaaaa
     3361 ctcatttgag ttaatgaggt aaagagaaaa tgagcaaaag cacaaacacg ctaagtgccg
     3421 gccgtccgag cgcacgcagc agcaaggctg caacgttggc cagcctggca gacacgccag
     3481 ccatgaagcg ggtcaacttt cagttgccgg cggaggatca caccaagctg aagatgtacg
     3541 cggtacgcca aggcaagacc attaccgagc tgctatctga atagatcgcg cagctaccag
     3601 agtaaatgag caaatgaata aatgagtaga tgaattttag cggctaaagg aggcggcatg
     3661 gaaaatcaag aacaaccagg caccgacgcc gtggaatgcc ccatgtgtgg aggaacgggc
     3721 ggttggccag gcgtaagcgg ctgggttgtc tgccggccct gcaatggcac tggaaccccc
     3781 aagcccgagg aatcggcgtg acggtcgcaa accatccggc ccggtacaaa tcggcgcggc
     3841 gctgggtgat gacctggtgg agaagttgaa ggccgcgcag gccgcccagc ggcaacgcat
     3901 cgaggcagaa gcacgccccg gtgaatcgtg gcaagcggcc gctgatcgaa tccgcaaaga
     3961 atcccggcaa ccgccggcag ccggtgcgcc gtcgattagg aagccgccca agggcgacga
     4021 gcaaccagat tttttcgttc cgatgctcta tgacgtgggc acccgcgata gtcgcagcat
     4081 catggacgtg gccgttttcc gtctgtcgaa gcgtgaccga cgagctggcg aggtgatccg
     4141 ctacgagctt ccagacgggc acgtagaggt ttccgcaggg ccggccggca tggccagtgt
     4201 gtgggattac gacctggtac tgatggcggt ttcccatcta accgaatcca tgaaccgata
     4261 ccgggaaggg aagggagaca agcccggccg cgtgttccgt ccacacgttg cggacgtact
     4321 caagttctgc cggcgagccg atggcggaaa gcagaaagac gacctggtag aaacctgcat
     4381 tcggttaaac accacgcacg ttgccatgca gcgtacgaag aaggccaaga acggccgcct
     4441 ggtgacggta tccgagggtg aagccttgat tagccgctac aagatcgtaa agagcgaaac
     4501 cgggcggccg gagtacatcg agatcgagct agctgattgg atgtaccgcg agatcacaga
     4561 aggcaagaac ccggacgtgc tgacggttca ccccgattac tttttgatcg atcccggcat
     4621 cggccgtttt ctctaccgcc tggcacgccg cgccgcaggc aaggcagaag ccagatggtt
     4681 gttcaagacg atctacgaac gcagtggcag cgccggagag ttcaagaagt tctgtttcac
     4741 cgtgcgcaag ctgatcgggt caaatgacct gccggagtac gatttgaagg aggaggcggg
     4801 gcaggctggc ccgatcctag tcatgcgcta ccgcaacctg atcgagggcg aagcatccgc
     4861 cggttcctaa tgtacggagc agatgctagg gcaaattgcc ctagcagggg aaaaaggtcg
     4921 aaaaggactc tttcctgtgg atagcacgta cattgggaac ccaaagccgt acattgggaa
     4981 ccggaacccg tacattggga acccaaagcc gtacattggg aaccggtcac acatgtaagt
     5041 gactgatata aaagagaaaa aaggcgattt ttccgcctaa aactctttaa aacttattaa
     5101 aactcttaaa acccgcctgg cctgtgcata actgtctggc cagcgcacag ccgaagagct
     5161 gcaaaaagcg cctacccttc ggtcgctgcg ctccctacgc cccgccgctt cgcgtcggcc
     5221 tatcgcggcc gctggccgct caaaaatggc tggcctacgg ccaggcaatc taccagggcg
     5281 cggacaagcc gcgccgtcgc cactcgaccg ccggcgccca catcaaggca ccctgcctcg
     5341 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggtg acggtcacag
     5401 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     5461 gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct
     5521 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     5581 gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga
     5641 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5701 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5761 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5821 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5881 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5941 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     6001 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6061 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6121 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6181 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6241 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6301 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6361 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6421 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgcattcta ggtgattaga
     6481 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
     6541 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
     6601 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
     6661 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
     6721 ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat
     6781 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
     6841 gagcgagtcg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
     6901 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
     6961 ctaatacctg gaatgctgtt ttccctggga tcgcagtggt gagtaaccat gcatcatcag
     7021 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     7081 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     7141 ctggcgcatc gggcttccca tacaatcggt agattgtcgc acctgattgc ccgacattat
     7201 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggccttg
     7261 agcaagacgt ttcccgttga atatggctca taacagaact tattatttcc ttcctctttt
     7321 ctacagtatt taaagatacc ccaagaagct aattataaca agacgaactc caattcactg
     7381 ttccttgcat tctaaaacct taaataccag aaaacagctt tttcaaagtt gttttcaaag
     7441 ttggcgtata acatagtatc gacggagccg attttgaaac cgcggtgatc acaggcagca
     7501 acgctctgtc atcgttacaa tcaacatgct accctccgcg agatcatccg tgtttcaaac
     7561 ccggcagctt agttgccgtt cttccgaata gcatcggtaa catgagcaaa gtctgccgcc
     7621 ttacaacggc tctcccgctg acgccgtccc ggactgatgg gctgcctgta tcgagt
//
