LOCUS       sequence-299492-        7286 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 7286)
  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 7286)
  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-2KN0091
FEATURES             Location/Qualifiers
     source          1..7286
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    183..207
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             319..831
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             892..915
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      938..1138
                     /label=gene 7 terminator
                     /note="transcription terminator for octopine-type Ti 
                     plasmid gene 7"
     misc_feature    1160..1184
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             2485..3114
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIGTHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3548..4615
                     /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      4681..4875
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(5111..5133)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5219..5359
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5274..5293
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5374..5391)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5545..6133)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5625..5644)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6221..7036)
                     /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(6221..7036)
                     /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     6944..6963
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 gatcatccgt gtttcaaacc cggcagctta gttgccgttc ttccgaatag catcggtaac
       61 atgagcaaag tctgccgcct tacaacggct ctcccgctga cgccgtcccg gactgatggg
      121 ctgcctgtat cgagtggtga ttttgtgccg agctgccggt cggggagctg ttggctggct
      181 ggtggcagga tatattgtgg tgtaaacata acaagcttcg tctcaggagg ttctgtgggt
      241 tttttgggta gaagaagaaa cccatagcta tataatgaga aacaggggca aatccccgaa
      301 aacaaataac gcgaagcaat ggtattcaca ctcgaagatt tcgttgggga ctggcgacag
      361 acagccggct acaacctgga ccaagtcctt gaacagggag gtgtgtccag tttgtttcag
      421 aatctcgggg tgtccgtaac tccgatccaa aggattgtcc tgagcggtga aaatgggctg
      481 aagatcgaca tccatgtcat catcccgtat gaaggtctga gcggcgacca aatgggccag
      541 atcgaaaaaa tttttaaggt ggtgtaccct gtggatgatc atcactttaa ggtgatcctg
      601 cactatggca cactggtaat cgacggggtt acgccgaaca tgatcgacta tttcggacgg
      661 ccgtatgaag gcatcgccgt gttcgacggc aaaaagatca ctgtaacagg gaccctgtgg
      721 aacggcaaca aaattatcga cgagcgcctg atcaaccccg acggctccct gctgttccga
      781 gtaaccatca acggagtgac cggctggcgg ctgtgcgaac gcattctggc ttcggattat
      841 aaggaccatg acggagacta taaggaccat gacctcgacg ctgcagcagc ggattataag
      901 gacgatgacg ataagtgagc ttgatccccc gtcgacagct agctatatca tcaatttatg
      961 tattacacat aatatcgcac tcagtctttc atctacggca atgtaccagc tgatataatc
     1021 agttattgaa atatttctga atttaaactt gcatcaataa atttatgttt ttgcttggac
     1081 tataatacct gacttgttat tttatcaata aatatttaaa ctatatttct ttcaagatcg
     1141 ctgtcatgag acgaagcttt gacaggatat attggcgggt aaacctaaga gaaaagagcg
     1201 tttattagaa taatcggata tttaaaaggg cgtgaaaagg tttatccgtt cgtccatttg
     1261 tatgtgcatg ccaaccacag ggttcccctc gggatcaaag tactttgatc caacccctcc
     1321 gctgctatag tgcagtcggc ttctgacgtt cagtgcagcc gtcatctgaa aacgacatgt
     1381 cgcacaagtc ctaagttacg cgacaggctg ccgccctgcc cttttcctgg cgttttcttg
     1441 tcgcgtgttt tagtcgcata aagtagaata cttgcgacta gaaccggaga cattacgcca
     1501 tgaacaagag cgccgccgct ggcctgctgg gctatgcccg cgtcagcacc gacgaccagg
     1561 acttgaccaa ccaacgggcc gaactgcacg cggccggctg caccaagctg ttttccgaga
     1621 agatcaccgg caccaggcgc gaccgcccgg agctggccag gatgcttgac cacctacgcc
     1681 ctggcgacgt tgtgacagtg accaggctag accgcctggc ccgcagcacc cgcgacctac
     1741 tggacattgc cgagcgcatc caggaggccg gcgcgggcct gcgtagcctg gcagagccgt
     1801 gggccgacac caccacgccg gccggccgca tggtgttgac cgtgttcgcc ggcattgccg
     1861 agttcgagcg ttccctaatc atcgaccgca cccggagcgg gcgcgaggcc gccaaggccc
     1921 gaggcgtgaa gtttggcccc cgccctaccc tcaccccggc acagatcgcg cacgcccgcg
     1981 agctgatcga ccaggaaggc cgcaccgtga aagaggcggc tgcactgctt ggcgtgcatc
     2041 gctcgaccct gtaccgcgca cttgagcgca gcgaggaagt gacgcccacc gaggccaggc
     2101 ggcgcggtgc cttccgtgag gacgcattga ccgaggccga cgccctggcg gccgccgaga
     2161 atgaacgcca agaggaacaa gcatgaaacc gcaccaggac ggccaggacg aaccgttttt
     2221 cattaccgaa gagatcgagg cggagatgat cgcggccggg tacgtgttcg agccgcccgc
     2281 gcacctctca accgtgcggc tgcatgaaat cctggccggt ttgtctgatg ccaagctggc
     2341 ggcctggccg gccagcttgg ccgctgaaga aaccgagcgc cgccgtctaa aaaggtgatg
     2401 tgtatttgag taaaacagct tgcgtcatgc ggtcgctgcg tatatgatcc gatgagtaaa
     2461 taaacaaata cgcaagggga acgcatgaag gttatcgctg tacttaacca gaaaggcggg
     2521 tcaggcaaga cgaccatcgg aacccatcta gcccgcgccc tgcaactcgc cggggccgat
     2581 gttctgttag tcgattccga tccccagggc agtgcccgcg attgggcggc cgtgcgggaa
     2641 gatcaaccgc taaccgttgt cggcatcgac cgcccgacga ttgaccgcga cgtgaaggcc
     2701 atcggccggc gcgacttcgt agtgatcgac ggagcgcccc aggcggcgga cttggctgtg
     2761 tccgcgatca aggcagccga cttcgtgctg attccggtgc agccaagccc ttacgacata
     2821 tgggccaccg ccgacctggt ggagctggtt aagcagcgca ttgaggtcac ggatggaagg
     2881 ctacaagcgg cctttgtcgt gtcgcgggcg atcaaaggca cgcgcatcgg cggtgaggtt
     2941 gccgaggcgc tggccgggta cgagctgccc attcttgagt cccgtatcac gcagcgcgtg
     3001 agctacccag gcactgccgc cgccggcaca accgttcttg aatcagaacc cgagggcgac
     3061 gctgcccgcg aggtccaggc gctggccgct gaaattaaat caaaactcat ttgagttaat
     3121 gaggtaaaga gaaaatgagc aaaagcacaa acacgctaag tgccggccgt ccgagcgcac
     3181 gcagcagcaa ggctgcaacg ttggccagcc tggcagacac gccagccatg aagcgggtca
     3241 actttcagtt gccggcggag gatcacacca agctgaagat gtacgcggta cgccaaggca
     3301 agaccattac cgagctgcta tctgaataga tcgcgcagct accagagtaa atgagcaaat
     3361 gaataaatga gtagatgaat tttagcggct aaaggaggcg gcatggaaaa tcaagaacaa
     3421 ccaggcaccg acgccgtgga atgccccatg tgtggaggaa cgggcggttg gccaggcgta
     3481 agcggctggg ttgtctgccg gccctgcaat ggcactggaa cccccaagcc cgaggaatcg
     3541 gcgtgacggt cgcaaaccat ccggcccggt acaaatcggc gcggcgctgg gtgatgacct
     3601 ggtggagaag ttgaaggccg cgcaggccgc ccagcggcaa cgcatcgagg cagaagcacg
     3661 ccccggtgaa tcgtggcaag cggccgctga tcgaatccgc aaagaatccc ggcaaccgcc
     3721 ggcagccggt gcgccgtcga ttaggaagcc gcccaagggc gacgagcaac cagatttttt
     3781 cgttccgatg ctctatgacg tgggcacccg cgatagtcgc agcatcatgg acgtggccgt
     3841 tttccgtctg tcgaagcgtg accgacgagc tggcgaggtg atccgctacg agcttccaga
     3901 cgggcacgta gaggtttccg cagggccggc cggcatggcc agtgtgtggg attacgacct
     3961 ggtactgatg gcggtttccc atctaaccga atccatgaac cgataccggg aagggaaggg
     4021 agacaagccc ggccgcgtgt tccgtccaca cgttgcggac gtactcaagt tctgccggcg
     4081 agccgatggc ggaaagcaga aagacgacct ggtagaaacc tgcattcggt taaacaccac
     4141 gcacgttgcc atgcagcgta cgaagaaggc caagaacggc cgcctggtga cggtatccga
     4201 gggtgaagcc ttgattagcc gctacaagat cgtaaagagc gaaaccgggc ggccggagta
     4261 catcgagatc gagctagctg attggatgta ccgcgagatc acagaaggca agaacccgga
     4321 cgtgctgacg gttcaccccg attacttttt gatcgatccc ggcatcggcc gttttctcta
     4381 ccgcctggca cgccgcgccg caggcaaggc agaagccaga tggttgttca agacgatcta
     4441 cgaacgcagt ggcagcgccg gagagttcaa gaagttctgt ttcaccgtgc gcaagctgat
     4501 cgggtcaaat gacctgccgg agtacgattt gaaggaggag gcggggcagg ctggcccgat
     4561 cctagtcatg cgctaccgca acctgatcga gggcgaagca tccgccggtt cctaatgtac
     4621 ggagcagatg ctagggcaaa ttgccctagc aggggaaaaa ggtcgaaaag gactctttcc
     4681 tgtggatagc acgtacattg ggaacccaaa gccgtacatt gggaaccgga acccgtacat
     4741 tgggaaccca aagccgtaca ttgggaaccg gtcacacatg taagtgactg atataaaaga
     4801 gaaaaaaggc gatttttccg cctaaaactc tttaaaactt attaaaactc ttaaaacccg
     4861 cctggcctgt gcataactgt ctggccagcg cacagccgaa gagctgcaaa aagcgcctac
     4921 ccttcggtcg ctgcgctccc tacgccccgc cgcttcgcgt cggcctatcg cggccgctgg
     4981 ccgctcaaaa atggctggcc tacggccagg caatctacca gggcgcggac aagccgcgcc
     5041 gtcgccactc gaccgccggc gcccacatca aggcaccctg cctcgcgcgt ttcggtgatg
     5101 acggtgaaaa cctctgacac atgcagctcc cggtgacggt cacagcttgt ctgtaagcgg
     5161 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg
     5221 cagccatgac ccagtcacgt agcgatagcg gagtgtatac tggcttaact atgcggcatc
     5281 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     5341 gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5401 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5461 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5521 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5581 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5641 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5701 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5761 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5821 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5881 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5941 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     6001 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6061 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6121 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6181 aaactcacgt taagggattt tggtcatgca ttctaggtga ttagaaaaac tcatcgagca
     6241 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
     6301 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
     6361 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
     6421 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
     6481 aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
     6541 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agtcgaaata
     6601 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
     6661 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
     6721 ctgttttccc tgggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
     6781 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
     6841 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
     6901 tcccatacaa tcggtagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
     6961 acccatataa atcagcatcc atgttggaat ttaatcgcgg ccttgagcaa gacgtttccc
     7021 gttgaatatg gctcataaca gaacttatta tttccttcct cttttctaca gtatttaaag
     7081 ataccccaag aagctaatta taacaagacg aactccaatt cactgttcct tgcattctaa
     7141 aaccttaaat accagaaaac agctttttca aagttgtttt caaagttggc gtataacata
     7201 gtatcgacgg agccgatttt gaaaccgcgg tgatcacagg cagcaacgct ctgtcatcgt
     7261 tacaatcaac atgctaccct ccgcga
//
