LOCUS       sequence-299494-        7400 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 7400)
  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 7400)
  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-2KN0093
FEATURES             Location/Qualifiers
     source          1..7400
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    141..165
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             391..903
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             964..987
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      1010..1210
                     /label=gene 7 terminator
                     /note="transcription terminator for octopine-type Ti 
                     plasmid gene 7"
     misc_feature    1232..1256
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             2557..3186
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIGTHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3620..4687
                     /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      4753..4947
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(5183..5205)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5291..5431
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5346..5365
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5446..5463)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5617..6205)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5697..5716)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6293..7108)
                     /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(6293..7108)
                     /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     7016..7035
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 ccgaatagca tcggtaacat gagcaaagtc tgccgcctta caacggctct cccgctgacg
       61 ccgtcccgga ctgatgggct gcctgtatcg agtggtgatt ttgtgccgag ctgccggtcg
      121 gggagctgtt ggctggctgg tggcaggata tattgtggtg taaacataac aagcttcgtc
      181 tcaggaggtt atcacacttg gtaatttaga agaagaaacc cataggttct gtgggttttt
      241 tgggtagaag aagaaaccca taggttctgt gggttttttg ggtagaagaa gaaacccata
      301 ggttctgtgg gttttttggg tagaagaaga aacccatagc tatataatga gaaacagggg
      361 caaatccccg aaaacaaata acgcgaagca atggtattca cactcgaaga tttcgttggg
      421 gactggcgac agacagccgg ctacaacctg gaccaagtcc ttgaacaggg aggtgtgtcc
      481 agtttgtttc agaatctcgg ggtgtccgta actccgatcc aaaggattgt cctgagcggt
      541 gaaaatgggc tgaagatcga catccatgtc atcatcccgt atgaaggtct gagcggcgac
      601 caaatgggcc agatcgaaaa aatttttaag gtggtgtacc ctgtggatga tcatcacttt
      661 aaggtgatcc tgcactatgg cacactggta atcgacgggg ttacgccgaa catgatcgac
      721 tatttcggac ggccgtatga aggcatcgcc gtgttcgacg gcaaaaagat cactgtaaca
      781 gggaccctgt ggaacggcaa caaaattatc gacgagcgcc tgatcaaccc cgacggctcc
      841 ctgctgttcc gagtaaccat caacggagtg accggctggc ggctgtgcga acgcattctg
      901 gcttcggatt ataaggacca tgacggagac tataaggacc atgacctcga cgctgcagca
      961 gcggattata aggacgatga cgataagtga gcttgatccc ccgtcgacag ctagctatat
     1021 catcaattta tgtattacac ataatatcgc actcagtctt tcatctacgg caatgtacca
     1081 gctgatataa tcagttattg aaatatttct gaatttaaac ttgcatcaat aaatttatgt
     1141 ttttgcttgg actataatac ctgacttgtt attttatcaa taaatattta aactatattt
     1201 ctttcaagat cgctgtcatg agacgaagct ttgacaggat atattggcgg gtaaacctaa
     1261 gagaaaagag cgtttattag aataatcgga tatttaaaag ggcgtgaaaa ggtttatccg
     1321 ttcgtccatt tgtatgtgca tgccaaccac agggttcccc tcgggatcaa agtactttga
     1381 tccaacccct ccgctgctat agtgcagtcg gcttctgacg ttcagtgcag ccgtcatctg
     1441 aaaacgacat gtcgcacaag tcctaagtta cgcgacaggc tgccgccctg cccttttcct
     1501 ggcgttttct tgtcgcgtgt tttagtcgca taaagtagaa tacttgcgac tagaaccgga
     1561 gacattacgc catgaacaag agcgccgccg ctggcctgct gggctatgcc cgcgtcagca
     1621 ccgacgacca ggacttgacc aaccaacggg ccgaactgca cgcggccggc tgcaccaagc
     1681 tgttttccga gaagatcacc ggcaccaggc gcgaccgccc ggagctggcc aggatgcttg
     1741 accacctacg ccctggcgac gttgtgacag tgaccaggct agaccgcctg gcccgcagca
     1801 cccgcgacct actggacatt gccgagcgca tccaggaggc cggcgcgggc ctgcgtagcc
     1861 tggcagagcc gtgggccgac accaccacgc cggccggccg catggtgttg accgtgttcg
     1921 ccggcattgc cgagttcgag cgttccctaa tcatcgaccg cacccggagc gggcgcgagg
     1981 ccgccaaggc ccgaggcgtg aagtttggcc cccgccctac cctcaccccg gcacagatcg
     2041 cgcacgcccg cgagctgatc gaccaggaag gccgcaccgt gaaagaggcg gctgcactgc
     2101 ttggcgtgca tcgctcgacc ctgtaccgcg cacttgagcg cagcgaggaa gtgacgccca
     2161 ccgaggccag gcggcgcggt gccttccgtg aggacgcatt gaccgaggcc gacgccctgg
     2221 cggccgccga gaatgaacgc caagaggaac aagcatgaaa ccgcaccagg acggccagga
     2281 cgaaccgttt ttcattaccg aagagatcga ggcggagatg atcgcggccg ggtacgtgtt
     2341 cgagccgccc gcgcacctct caaccgtgcg gctgcatgaa atcctggccg gtttgtctga
     2401 tgccaagctg gcggcctggc cggccagctt ggccgctgaa gaaaccgagc gccgccgtct
     2461 aaaaaggtga tgtgtatttg agtaaaacag cttgcgtcat gcggtcgctg cgtatatgat
     2521 ccgatgagta aataaacaaa tacgcaaggg gaacgcatga aggttatcgc tgtacttaac
     2581 cagaaaggcg ggtcaggcaa gacgaccatc ggaacccatc tagcccgcgc cctgcaactc
     2641 gccggggccg atgttctgtt agtcgattcc gatccccagg gcagtgcccg cgattgggcg
     2701 gccgtgcggg aagatcaacc gctaaccgtt gtcggcatcg accgcccgac gattgaccgc
     2761 gacgtgaagg ccatcggccg gcgcgacttc gtagtgatcg acggagcgcc ccaggcggcg
     2821 gacttggctg tgtccgcgat caaggcagcc gacttcgtgc tgattccggt gcagccaagc
     2881 ccttacgaca tatgggccac cgccgacctg gtggagctgg ttaagcagcg cattgaggtc
     2941 acggatggaa ggctacaagc ggcctttgtc gtgtcgcggg cgatcaaagg cacgcgcatc
     3001 ggcggtgagg ttgccgaggc gctggccggg tacgagctgc ccattcttga gtcccgtatc
     3061 acgcagcgcg tgagctaccc aggcactgcc gccgccggca caaccgttct tgaatcagaa
     3121 cccgagggcg acgctgcccg cgaggtccag gcgctggccg ctgaaattaa atcaaaactc
     3181 atttgagtta atgaggtaaa gagaaaatga gcaaaagcac aaacacgcta agtgccggcc
     3241 gtccgagcgc acgcagcagc aaggctgcaa cgttggccag cctggcagac acgccagcca
     3301 tgaagcgggt caactttcag ttgccggcgg aggatcacac caagctgaag atgtacgcgg
     3361 tacgccaagg caagaccatt accgagctgc tatctgaata gatcgcgcag ctaccagagt
     3421 aaatgagcaa atgaataaat gagtagatga attttagcgg ctaaaggagg cggcatggaa
     3481 aatcaagaac aaccaggcac cgacgccgtg gaatgcccca tgtgtggagg aacgggcggt
     3541 tggccaggcg taagcggctg ggttgtctgc cggccctgca atggcactgg aacccccaag
     3601 cccgaggaat cggcgtgacg gtcgcaaacc atccggcccg gtacaaatcg gcgcggcgct
     3661 gggtgatgac ctggtggaga agttgaaggc cgcgcaggcc gcccagcggc aacgcatcga
     3721 ggcagaagca cgccccggtg aatcgtggca agcggccgct gatcgaatcc gcaaagaatc
     3781 ccggcaaccg ccggcagccg gtgcgccgtc gattaggaag ccgcccaagg gcgacgagca
     3841 accagatttt ttcgttccga tgctctatga cgtgggcacc cgcgatagtc gcagcatcat
     3901 ggacgtggcc gttttccgtc tgtcgaagcg tgaccgacga gctggcgagg tgatccgcta
     3961 cgagcttcca gacgggcacg tagaggtttc cgcagggccg gccggcatgg ccagtgtgtg
     4021 ggattacgac ctggtactga tggcggtttc ccatctaacc gaatccatga accgataccg
     4081 ggaagggaag ggagacaagc ccggccgcgt gttccgtcca cacgttgcgg acgtactcaa
     4141 gttctgccgg cgagccgatg gcggaaagca gaaagacgac ctggtagaaa cctgcattcg
     4201 gttaaacacc acgcacgttg ccatgcagcg tacgaagaag gccaagaacg gccgcctggt
     4261 gacggtatcc gagggtgaag ccttgattag ccgctacaag atcgtaaaga gcgaaaccgg
     4321 gcggccggag tacatcgaga tcgagctagc tgattggatg taccgcgaga tcacagaagg
     4381 caagaacccg gacgtgctga cggttcaccc cgattacttt ttgatcgatc ccggcatcgg
     4441 ccgttttctc taccgcctgg cacgccgcgc cgcaggcaag gcagaagcca gatggttgtt
     4501 caagacgatc tacgaacgca gtggcagcgc cggagagttc aagaagttct gtttcaccgt
     4561 gcgcaagctg atcgggtcaa atgacctgcc ggagtacgat ttgaaggagg aggcggggca
     4621 ggctggcccg atcctagtca tgcgctaccg caacctgatc gagggcgaag catccgccgg
     4681 ttcctaatgt acggagcaga tgctagggca aattgcccta gcaggggaaa aaggtcgaaa
     4741 aggactcttt cctgtggata gcacgtacat tgggaaccca aagccgtaca ttgggaaccg
     4801 gaacccgtac attgggaacc caaagccgta cattgggaac cggtcacaca tgtaagtgac
     4861 tgatataaaa gagaaaaaag gcgatttttc cgcctaaaac tctttaaaac ttattaaaac
     4921 tcttaaaacc cgcctggcct gtgcataact gtctggccag cgcacagccg aagagctgca
     4981 aaaagcgcct acccttcggt cgctgcgctc cctacgcccc gccgcttcgc gtcggcctat
     5041 cgcggccgct ggccgctcaa aaatggctgg cctacggcca ggcaatctac cagggcgcgg
     5101 acaagccgcg ccgtcgccac tcgaccgccg gcgcccacat caaggcaccc tgcctcgcgc
     5161 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggtgacg gtcacagctt
     5221 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     5281 ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa
     5341 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     5401 cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc
     5461 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5521 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5581 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5641 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5701 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5761 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5821 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5881 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5941 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6001 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     6061 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6121 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6181 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6241 tcagtggaac gaaaactcac gttaagggat tttggtcatg cattctaggt gattagaaaa
     6301 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt
     6361 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
     6421 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
     6481 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg
     6541 gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac
     6601 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag
     6661 cgagtcgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc
     6721 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta
     6781 atacctggaa tgctgttttc cctgggatcg cagtggtgag taaccatgca tcatcaggag
     6841 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
     6901 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
     6961 gcgcatcggg cttcccatac aatcggtaga ttgtcgcacc tgattgcccg acattatcgc
     7021 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggccttgagc
     7081 aagacgtttc ccgttgaata tggctcataa cagaacttat tatttccttc ctcttttcta
     7141 cagtatttaa agatacccca agaagctaat tataacaaga cgaactccaa ttcactgttc
     7201 cttgcattct aaaaccttaa ataccagaaa acagcttttt caaagttgtt ttcaaagttg
     7261 gcgtataaca tagtatcgac ggagccgatt ttgaaaccgc ggtgatcaca ggcagcaacg
     7321 ctctgtcatc gttacaatca acatgctacc ctccgcgaga tcatccgtgt ttcaaacccg
     7381 gcagcttagt tgccgttctt
//
