LOCUS       sequence-299496-        7558 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 7558)
  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 7558)
  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-2KN0101
FEATURES             Location/Qualifiers
     source          1..7558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      279..473
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(709..731)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    817..957
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     872..891
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(972..989)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1143..1731)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1223..1242)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1819..2634)
                     /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(1819..2634)
                     /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     2542..2561
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    3067..3091
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     protein_bind    3126..3159
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    3185..3218
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    3248..3281
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    3307..3340
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        3357..3403
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             3479..3991
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             4052..4075
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      4098..4298
                     /label=gene 7 terminator
                     /note="transcription terminator for octopine-type Ti 
                     plasmid gene 7"
     misc_feature    4316..4340
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             5641..6270
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIGTHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             join(6704..7558,1..213)
                     /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"
ORIGIN
        1 ggcaaggcag aagccagatg gttgttcaag acgatctacg aacgcagtgg cagcgccgga
       61 gagttcaaga agttctgttt caccgtgcgc aagctgatcg ggtcaaatga cctgccggag
      121 tacgatttga aggaggaggc ggggcaggct ggcccgatcc tagtcatgcg ctaccgcaac
      181 ctgatcgagg gcgaagcatc cgccggttcc taatgtacgg agcagatgct agggcaaatt
      241 gccctagcag gggaaaaagg tcgaaaagga ctctttcctg tggatagcac gtacattggg
      301 aacccaaagc cgtacattgg gaaccggaac ccgtacattg ggaacccaaa gccgtacatt
      361 gggaaccggt cacacatgta agtgactgat ataaaagaga aaaaaggcga tttttccgcc
      421 taaaactctt taaaacttat taaaactctt aaaacccgcc tggcctgtgc ataactgtct
      481 ggccagcgca cagccgaaga gctgcaaaaa gcgcctaccc ttcggtcgct gcgctcccta
      541 cgccccgccg cttcgcgtcg gcctatcgcg gccgctggcc gctcaaaaat ggctggccta
      601 cggccaggca atctaccagg gcgcggacaa gccgcgccgt cgccactcga ccgccggcgc
      661 ccacatcaag gcaccctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
      721 gcagctcccg gtgacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
      781 tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag
      841 cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg
      901 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
      961 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     1021 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     1081 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     1141 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     1201 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     1261 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     1321 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     1381 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     1441 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     1501 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     1561 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     1621 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     1681 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     1741 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     1801 gtcatgcatt ctaggtgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
     1861 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     1921 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     1981 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     2041 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
     2101 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     2161 gttattcatt cgtgattgcg cctgagcgag tcgaaatacg cgatcgctgt taaaaggaca
     2221 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     2281 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttccctg ggatcgcagt
     2341 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     2401 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     2461 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc ggtagattgt
     2521 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     2581 gttggaattt aatcgcggcc ttgagcaaga cgtttcccgt tgaatatggc tcataacaga
     2641 acttattatt tccttcctct tttctacagt atttaaagat accccaagaa gctaattata
     2701 acaagacgaa ctccaattca ctgttccttg cattctaaaa ccttaaatac cagaaaacag
     2761 ctttttcaaa gttgttttca aagttggcgt ataacatagt atcgacggag ccgattttga
     2821 aaccgcggtg atcacaggca gcaacgctct gtcatcgtta caatcaacat gctaccctcc
     2881 gcgagatcat ccgtgtttca aacccggcag cttagttgcc gttcttccga atagcatcgg
     2941 taacatgagc aaagtctgcc gccttacaac ggctctcccg ctgacgccgt cccggactga
     3001 tgggctgcct gtatcgagtg gtgattttgt gccgagctgc cggtcgggga gctgttggct
     3061 ggctggtggc aggatatatt gtggtgtaaa cataacaagc ttcgtctcag tcaggagcgg
     3121 tgcgggtgcc agggcgtgcc cttgggctcc ccgggcgcgt actccactcc ccgagccggt
     3181 gcgggtgcca gggcgtgccc ttgggctccc cgggcgcgta ctccactccc cccatggtgc
     3241 ggtgcgggtg ccagggcgtg cccttgggct ccccgggcgc gtactccact ccccgagccg
     3301 gtgcgggtgc cagggcgtgc ccttgggctc cccgggcgcg tactccactc cccttcgcaa
     3361 gacccttcct ctatataagg aagttcattt catttggaga ggactccggt atttttacaa
     3421 caattaccac aacaaaacaa acaacaaaca acattacaat ttactattct agtcgaaaat
     3481 ggtattcaca ctcgaagatt tcgttgggga ctggcgacag acagccggct acaacctgga
     3541 ccaagtcctt gaacagggag gtgtgtccag tttgtttcag aatctcgggg tgtccgtaac
     3601 tccgatccaa aggattgtcc tgagcggtga aaatgggctg aagatcgaca tccatgtcat
     3661 catcccgtat gaaggtctga gcggcgacca aatgggccag atcgaaaaaa tttttaaggt
     3721 ggtgtaccct gtggatgatc atcactttaa ggtgatcctg cactatggca cactggtaat
     3781 cgacggggtt acgccgaaca tgatcgacta tttcggacgg ccgtatgaag gcatcgccgt
     3841 gttcgacggc aaaaagatca ctgtaacagg gaccctgtgg aacggcaaca aaattatcga
     3901 cgagcgcctg atcaaccccg acggctccct gctgttccga gtaaccatca acggagtgac
     3961 cggctggcgg ctgtgcgaac gcattctggc ttcggattat aaggaccatg acggagacta
     4021 taaggaccat gacctcgacg ctgcagcagc ggattataag gacgatgacg ataagtgagc
     4081 ttgatccccc gtcgacagct agctatatca tcaatttatg tattacacat aatatcgcac
     4141 tcagtctttc atctacggca atgtaccagc tgatataatc agttattgaa atatttctga
     4201 atttaaactt gcatcaataa atttatgttt ttgcttggac tataatacct gacttgttat
     4261 tttatcaata aatatttaaa ctatatttct ttcaagatcg cttgagacga agctttgaca
     4321 ggatatattg gcgggtaaac ctaagagaaa agagcgttta ttagaataat cggatattta
     4381 aaagggcgtg aaaaggttta tccgttcgtc catttgtatg tgcatgccaa ccacagggtt
     4441 cccctcggga tcaaagtact ttgatccaac ccctccgctg ctatagtgca gtcggcttct
     4501 gacgttcagt gcagccgtca tctgaaaacg acatgtcgca caagtcctaa gttacgcgac
     4561 aggctgccgc cctgcccttt tcctggcgtt ttcttgtcgc gtgttttagt cgcataaagt
     4621 agaatacttg cgactagaac cggagacatt acgccatgaa caagagcgcc gccgctggcc
     4681 tgctgggcta tgcccgcgtc agcaccgacg accaggactt gaccaaccaa cgggccgaac
     4741 tgcacgcggc cggctgcacc aagctgtttt ccgagaagat caccggcacc aggcgcgacc
     4801 gcccggagct ggccaggatg cttgaccacc tacgccctgg cgacgttgtg acagtgacca
     4861 ggctagaccg cctggcccgc agcacccgcg acctactgga cattgccgag cgcatccagg
     4921 aggccggcgc gggcctgcgt agcctggcag agccgtgggc cgacaccacc acgccggccg
     4981 gccgcatggt gttgaccgtg ttcgccggca ttgccgagtt cgagcgttcc ctaatcatcg
     5041 accgcacccg gagcgggcgc gaggccgcca aggcccgagg cgtgaagttt ggcccccgcc
     5101 ctaccctcac cccggcacag atcgcgcacg cccgcgagct gatcgaccag gaaggccgca
     5161 ccgtgaaaga ggcggctgca ctgcttggcg tgcatcgctc gaccctgtac cgcgcacttg
     5221 agcgcagcga ggaagtgacg cccaccgagg ccaggcggcg cggtgccttc cgtgaggacg
     5281 cattgaccga ggccgacgcc ctggcggccg ccgagaatga acgccaagag gaacaagcat
     5341 gaaaccgcac caggacggcc aggacgaacc gtttttcatt accgaagaga tcgaggcgga
     5401 gatgatcgcg gccgggtacg tgttcgagcc gcccgcgcac ctctcaaccg tgcggctgca
     5461 tgaaatcctg gccggtttgt ctgatgccaa gctggcggcc tggccggcca gcttggccgc
     5521 tgaagaaacc gagcgccgcc gtctaaaaag gtgatgtgta tttgagtaaa acagcttgcg
     5581 tcatgcggtc gctgcgtata tgatccgatg agtaaataaa caaatacgca aggggaacgc
     5641 atgaaggtta tcgctgtact taaccagaaa ggcgggtcag gcaagacgac catcggaacc
     5701 catctagccc gcgccctgca actcgccggg gccgatgttc tgttagtcga ttccgatccc
     5761 cagggcagtg cccgcgattg ggcggccgtg cgggaagatc aaccgctaac cgttgtcggc
     5821 atcgaccgcc cgacgattga ccgcgacgtg aaggccatcg gccggcgcga cttcgtagtg
     5881 atcgacggag cgccccaggc ggcggacttg gctgtgtccg cgatcaaggc agccgacttc
     5941 gtgctgattc cggtgcagcc aagcccttac gacatatggg ccaccgccga cctggtggag
     6001 ctggttaagc agcgcattga ggtcacggat ggaaggctac aagcggcctt tgtcgtgtcg
     6061 cgggcgatca aaggcacgcg catcggcggt gaggttgccg aggcgctggc cgggtacgag
     6121 ctgcccattc ttgagtcccg tatcacgcag cgcgtgagct acccaggcac tgccgccgcc
     6181 ggcacaaccg ttcttgaatc agaacccgag ggcgacgctg cccgcgaggt ccaggcgctg
     6241 gccgctgaaa ttaaatcaaa actcatttga gttaatgagg taaagagaaa atgagcaaaa
     6301 gcacaaacac gctaagtgcc ggccgtccga gcgcacgcag cagcaaggct gcaacgttgg
     6361 ccagcctggc agacacgcca gccatgaagc gggtcaactt tcagttgccg gcggaggatc
     6421 acaccaagct gaagatgtac gcggtacgcc aaggcaagac cattaccgag ctgctatctg
     6481 aatagatcgc gcagctacca gagtaaatga gcaaatgaat aaatgagtag atgaatttta
     6541 gcggctaaag gaggcggcat ggaaaatcaa gaacaaccag gcaccgacgc cgtggaatgc
     6601 cccatgtgtg gaggaacggg cggttggcca ggcgtaagcg gctgggttgt ctgccggccc
     6661 tgcaatggca ctggaacccc caagcccgag gaatcggcgt gacggtcgca aaccatccgg
     6721 cccggtacaa atcggcgcgg cgctgggtga tgacctggtg gagaagttga aggccgcgca
     6781 ggccgcccag cggcaacgca tcgaggcaga agcacgcccc ggtgaatcgt ggcaagcggc
     6841 cgctgatcga atccgcaaag aatcccggca accgccggca gccggtgcgc cgtcgattag
     6901 gaagccgccc aagggcgacg agcaaccaga ttttttcgtt ccgatgctct atgacgtggg
     6961 cacccgcgat agtcgcagca tcatggacgt ggccgttttc cgtctgtcga agcgtgaccg
     7021 acgagctggc gaggtgatcc gctacgagct tccagacggg cacgtagagg tttccgcagg
     7081 gccggccggc atggccagtg tgtgggatta cgacctggta ctgatggcgg tttcccatct
     7141 aaccgaatcc atgaaccgat accgggaagg gaagggagac aagcccggcc gcgtgttccg
     7201 tccacacgtt gcggacgtac tcaagttctg ccggcgagcc gatggcggaa agcagaaaga
     7261 cgacctggta gaaacctgca ttcggttaaa caccacgcac gttgccatgc agcgtacgaa
     7321 gaaggccaag aacggccgcc tggtgacggt atccgagggt gaagccttga ttagccgcta
     7381 caagatcgta aagagcgaaa ccgggcggcc ggagtacatc gagatcgagc tagctgattg
     7441 gatgtaccgc gagatcacag aaggcaagaa cccggacgtg ctgacggttc accccgatta
     7501 ctttttgatc gatcccggca tcggccgttt tctctaccgc ctggcacgcc gcgccgca
//
