LOCUS       sequence-299493-        7323 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 7323)
  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 7323)
  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-2KN0092
FEATURES             Location/Qualifiers
     source          1..7323
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    179..203
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             352..864
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             925..948
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      971..1171
                     /label=gene 7 terminator
                     /note="transcription terminator for octopine-type Ti 
                     plasmid gene 7"
     misc_feature    1193..1217
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             2518..3147
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIGTHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3581..4648
                     /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      4714..4908
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(5144..5166)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5252..5392
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5307..5326
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5407..5424)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5578..6166)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5658..5677)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6254..7069)
                     /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(6254..7069)
                     /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     6977..6996
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 atccgtgttt caaacccggc agcttagttg ccgttcttcc gaatagcatc ggtaacatga
       61 gcaaagtctg ccgccttaca acggctctcc cgctgacgcc gtcccggact gatgggctgc
      121 ctgtatcgag tggtgatttt gtgccgagct gccggtcggg gagctgttgg ctggctggtg
      181 gcaggatata ttgtggtgta aactaacaag cttcgtctca ggaggttatc acacttggta
      241 atttagaaga agaaacccat aggttctgtg ggttttttgg gtagaagaag aaacccatag
      301 ctatataatg agaaacaggg gcaaatcccc gaaaacaaat aacgcgaagc aatggtattc
      361 acactcgaag atttcgttgg ggactggcga cagacagccg gctacaacct ggaccaagtc
      421 cttgaacagg gaggtgtgtc cagtttgttt cagaatctcg gggtgtccgt aactccgatc
      481 caaaggattg tcctgagcgg tgaaaatggg ctgaagatcg acatccatgt catcatcccg
      541 tatgaaggtc tgagcggcga ccaaatgggc cagatcgaaa aaatttttaa ggtggtgtac
      601 cctgtggatg atcatcactt taaggtgatc ctgcactatg gcacactggt aatcgacggg
      661 gttacgccga acatgatcga ctatttcgga cggccgtatg aaggcatcgc cgtgttcgac
      721 ggcaaaaaga tcactgtaac agggaccctg tggaacggca acaaaattat cgacgagcgc
      781 ctgatcaacc ccgacggctc cctgctgttc cgagtaacca tcaacggagt gaccggctgg
      841 cggctgtgcg aacgcattct ggcttcggat tataaggacc atgacggaga ctataaggac
      901 catgacctcg acgctgcagc agcggattat aaggacgatg acgataagtg agcttgatcc
      961 cccgtcgaca gctagctata tcatcaattt atgtattaca cataatatcg cactcagtct
     1021 ttcatctacg gcaatgtacc agctgatata atcagttatt gaaatatttc tgaatttaaa
     1081 cttgcatcaa taaatttatg tttttgcttg gactataata cctgacttgt tattttatca
     1141 ataaatattt aaactatatt tctttcaaga tcgctgtcat gagacgaagc tttgacagga
     1201 tatattggcg ggtaaaccta agagaaaaga gcgtttatta gaataatcgg atatttaaaa
     1261 gggcgtgaaa aggtttatcc gttcgtccat ttgtatgtgc atgccaacca cagggttccc
     1321 ctcgggatca aagtactttg atccaacccc tccgctgcta tagtgcagtc ggcttctgac
     1381 gttcagtgca gccgtcatct gaaaacgaca tgtcgcacaa gtcctaagtt acgcgacagg
     1441 ctgccgccct gcccttttcc tggcgttttc ttgtcgcgtg ttttagtcgc ataaagtaga
     1501 atacttgcga ctagaaccgg agacattacg ccatgaacaa gagcgccgcc gctggcctgc
     1561 tgggctatgc ccgcgtcagc accgacgacc aggacttgac caaccaacgg gccgaactgc
     1621 acgcggccgg ctgcaccaag ctgttttccg agaagatcac cggcaccagg cgcgaccgcc
     1681 cggagctggc caggatgctt gaccacctac gccctggcga cgttgtgaca gtgaccaggc
     1741 tagaccgcct ggcccgcagc acccgcgacc tactggacat tgccgagcgc atccaggagg
     1801 ccggcgcggg cctgcgtagc ctggcagagc cgtgggccga caccaccacg ccggccggcc
     1861 gcatggtgtt gaccgtgttc gccggcattg ccgagttcga gcgttcccta atcatcgacc
     1921 gcacccggag cgggcgcgag gccgccaagg cccgaggcgt gaagtttggc ccccgcccta
     1981 ccctcacccc ggcacagatc gcgcacgccc gcgagctgat cgaccaggaa ggccgcaccg
     2041 tgaaagaggc ggctgcactg cttggcgtgc atcgctcgac cctgtaccgc gcacttgagc
     2101 gcagcgagga agtgacgccc accgaggcca ggcggcgcgg tgccttccgt gaggacgcat
     2161 tgaccgaggc cgacgccctg gcggccgccg agaatgaacg ccaagaggaa caagcatgaa
     2221 accgcaccag gacggccagg acgaaccgtt tttcattacc gaagagatcg aggcggagat
     2281 gatcgcggcc gggtacgtgt tcgagccgcc cgcgcacctc tcaaccgtgc ggctgcatga
     2341 aatcctggcc ggtttgtctg atgccaagct ggcggcctgg ccggccagct tggccgctga
     2401 agaaaccgag cgccgccgtc taaaaaggtg atgtgtattt gagtaaaaca gcttgcgtca
     2461 tgcggtcgct gcgtatatga tccgatgagt aaataaacaa atacgcaagg ggaacgcatg
     2521 aaggttatcg ctgtacttaa ccagaaaggc gggtcaggca agacgaccat cggaacccat
     2581 ctagcccgcg ccctgcaact cgccggggcc gatgttctgt tagtcgattc cgatccccag
     2641 ggcagtgccc gcgattgggc ggccgtgcgg gaagatcaac cgctaaccgt tgtcggcatc
     2701 gaccgcccga cgattgaccg cgacgtgaag gccatcggcc ggcgcgactt cgtagtgatc
     2761 gacggagcgc cccaggcggc ggacttggct gtgtccgcga tcaaggcagc cgacttcgtg
     2821 ctgattccgg tgcagccaag cccttacgac atatgggcca ccgccgacct ggtggagctg
     2881 gttaagcagc gcattgaggt cacggatgga aggctacaag cggcctttgt cgtgtcgcgg
     2941 gcgatcaaag gcacgcgcat cggcggtgag gttgccgagg cgctggccgg gtacgagctg
     3001 cccattcttg agtcccgtat cacgcagcgc gtgagctacc caggcactgc cgccgccggc
     3061 acaaccgttc ttgaatcaga acccgagggc gacgctgccc gcgaggtcca ggcgctggcc
     3121 gctgaaatta aatcaaaact catttgagtt aatgaggtaa agagaaaatg agcaaaagca
     3181 caaacacgct aagtgccggc cgtccgagcg cacgcagcag caaggctgca acgttggcca
     3241 gcctggcaga cacgccagcc atgaagcggg tcaactttca gttgccggcg gaggatcaca
     3301 ccaagctgaa gatgtacgcg gtacgccaag gcaagaccat taccgagctg ctatctgaat
     3361 agatcgcgca gctaccagag taaatgagca aatgaataaa tgagtagatg aattttagcg
     3421 gctaaaggag gcggcatgga aaatcaagaa caaccaggca ccgacgccgt ggaatgcccc
     3481 atgtgtggag gaacgggcgg ttggccaggc gtaagcggct gggttgtctg ccggccctgc
     3541 aatggcactg gaacccccaa gcccgaggaa tcggcgtgac ggtcgcaaac catccggccc
     3601 ggtacaaatc ggcgcggcgc tgggtgatga cctggtggag aagttgaagg ccgcgcaggc
     3661 cgcccagcgg caacgcatcg aggcagaagc acgccccggt gaatcgtggc aagcggccgc
     3721 tgatcgaatc cgcaaagaat cccggcaacc gccggcagcc ggtgcgccgt cgattaggaa
     3781 gccgcccaag ggcgacgagc aaccagattt tttcgttccg atgctctatg acgtgggcac
     3841 ccgcgatagt cgcagcatca tggacgtggc cgttttccgt ctgtcgaagc gtgaccgacg
     3901 agctggcgag gtgatccgct acgagcttcc agacgggcac gtagaggttt ccgcagggcc
     3961 ggccggcatg gccagtgtgt gggattacga cctggtactg atggcggttt cccatctaac
     4021 cgaatccatg aaccgatacc gggaagggaa gggagacaag cccggccgcg tgttccgtcc
     4081 acacgttgcg gacgtactca agttctgccg gcgagccgat ggcggaaagc agaaagacga
     4141 cctggtagaa acctgcattc ggttaaacac cacgcacgtt gccatgcagc gtacgaagaa
     4201 ggccaagaac ggccgcctgg tgacggtatc cgagggtgaa gccttgatta gccgctacaa
     4261 gatcgtaaag agcgaaaccg ggcggccgga gtacatcgag atcgagctag ctgattggat
     4321 gtaccgcgag atcacagaag gcaagaaccc ggacgtgctg acggttcacc ccgattactt
     4381 tttgatcgat cccggcatcg gccgttttct ctaccgcctg gcacgccgcg ccgcaggcaa
     4441 ggcagaagcc agatggttgt tcaagacgat ctacgaacgc agtggcagcg ccggagagtt
     4501 caagaagttc tgtttcaccg tgcgcaagct gatcgggtca aatgacctgc cggagtacga
     4561 tttgaaggag gaggcggggc aggctggccc gatcctagtc atgcgctacc gcaacctgat
     4621 cgagggcgaa gcatccgccg gttcctaatg tacggagcag atgctagggc aaattgccct
     4681 agcaggggaa aaaggtcgaa aaggactctt tcctgtggat agcacgtaca ttgggaaccc
     4741 aaagccgtac attgggaacc ggaacccgta cattgggaac ccaaagccgt acattgggaa
     4801 ccggtcacac atgtaagtga ctgatataaa agagaaaaaa ggcgattttt ccgcctaaaa
     4861 ctctttaaaa cttattaaaa ctcttaaaac ccgcctggcc tgtgcataac tgtctggcca
     4921 gcgcacagcc gaagagctgc aaaaagcgcc tacccttcgg tcgctgcgct ccctacgccc
     4981 cgccgcttcg cgtcggccta tcgcggccgc tggccgctca aaaatggctg gcctacggcc
     5041 aggcaatcta ccagggcgcg gacaagccgc gccgtcgcca ctcgaccgcc ggcgcccaca
     5101 tcaaggcacc ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     5161 tcccggtgac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     5221 gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata
     5281 gcggagtgta tactggctta actatgcggc atcagagcag attgtactga gagtgcacca
     5341 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc
     5401 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5461 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5521 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5581 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5641 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5701 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5761 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5821 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5881 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5941 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6001 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6061 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6121 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6181 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6241 gcattctagg tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat
     6301 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac
     6361 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa
     6421 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac
     6481 catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt
     6541 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat
     6601 tcattcgtga ttgcgcctga gcgagtcgaa atacgcgatc gctgttaaaa ggacaattac
     6661 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
     6721 ctgaatcagg atattcttct aatacctgga atgctgtttt ccctgggatc gcagtggtga
     6781 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
     6841 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc
     6901 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcggtag attgtcgcac
     6961 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
     7021 aatttaatcg cggccttgag caagacgttt cccgttgaat atggctcata acagaactta
     7081 ttatttcctt cctcttttct acagtattta aagatacccc aagaagctaa ttataacaag
     7141 acgaactcca attcactgtt ccttgcattc taaaacctta aataccagaa aacagctttt
     7201 tcaaagttgt tttcaaagtt ggcgtataac atagtatcga cggagccgat tttgaaaccg
     7261 cggtgatcac aggcagcaac gctctgtcat cgttacaatc aacatgctac cctccgcgag
     7321 atc
//
