LOCUS       sequence-299495-        7440 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 7440)
  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 7440)
  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-2KN0100
FEATURES             Location/Qualifiers
     source          1..7440
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     4..23
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    529..553
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     protein_bind    592..625
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    651..684
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        701..747
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             823..1335
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             1396..1419
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      1442..1642
                     /label=gene 7 terminator
                     /note="transcription terminator for octopine-type Ti 
                     plasmid gene 7"
     misc_feature    1660..1684
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             2985..3614
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIGTHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4048..5115
                     /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      5181..5375
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(5611..5633)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5719..5859
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5774..5793
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5874..5891)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6045..6633)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6125..6144)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6721..7440,1..96))
                     /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(join(6721..7440,1..96))
                     /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"
ORIGIN
        1 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
       61 ccttgagcaa gacgtttccc gttgaatatg gctcataaca gaacttatta tttccttcct
      121 cttttctaca gtatttaaag ataccccaag aagctaatta taacaagacg aactccaatt
      181 cactgttcct tgcattctaa aaccttaaat accagaaaac agctttttca aagttgtttt
      241 caaagttggc gtataacata gtatcgacgg agccgatttt gaaaccgcgg tgatcacagg
      301 cagcaacgct ctgtcatcgt tacaatcaac atgctaccct ccgcgagatc atccgtgttt
      361 caaacccggc agcttagttg ccgttcttcc gaatagcatc ggtaacatga gcaaagtctg
      421 ccgccttaca acggctctcc cgctgacgcc gtcccggact gatgggctgc ctgtatcgag
      481 tggtgatttt gtgccgagct gccggtcggg gagctgttgg ctggctggtg gcaggatata
      541 ttgtggtgta aacataacaa gcttcgtctc agtcaggagg gagcggtgcg ggtgccaggg
      601 cgtgcccttg ggctccccgg gcgcgtactc cactccccga gccggtgcgg gtgccagggc
      661 gtgcccttgg gctccccggg cgcgtactcc actccccttc gcaagaccct tcctctatat
      721 aaggaagttc atttcatttg gagaggactc cggtattttt acaacaatta ccacaacaaa
      781 acaaacaaca aacaacatta caatttacta ttctagtcga aaatggtatt cacactcgaa
      841 gatttcgttg gggactggcg acagacagcc ggctacaacc tggaccaagt ccttgaacag
      901 ggaggtgtgt ccagtttgtt tcagaatctc ggggtgtccg taactccgat ccaaaggatt
      961 gtcctgagcg gtgaaaatgg gctgaagatc gacatccatg tcatcatccc gtatgaaggt
     1021 ctgagcggcg accaaatggg ccagatcgaa aaaattttta aggtggtgta ccctgtggat
     1081 gatcatcact ttaaggtgat cctgcactat ggcacactgg taatcgacgg ggttacgccg
     1141 aacatgatcg actatttcgg acggccgtat gaaggcatcg ccgtgttcga cggcaaaaag
     1201 atcactgtaa cagggaccct gtggaacggc aacaaaatta tcgacgagcg cctgatcaac
     1261 cccgacggct ccctgctgtt ccgagtaacc atcaacggag tgaccggctg gcggctgtgc
     1321 gaacgcattc tggcttcgga ttataaggac catgacggag actataagga ccatgacctc
     1381 gacgctgcag cagcggatta taaggacgat gacgataagt gagcttgatc ccccgtcgac
     1441 agctagctat atcatcaatt tatgtattac acataatatc gcactcagtc tttcatctac
     1501 ggcaatgtac cagctgatat aatcagttat tgaaatattt ctgaatttaa acttgcatca
     1561 ataaatttat gtttttgctt ggactataat acctgacttg ttattttatc aataaatatt
     1621 taaactatat ttctttcaag atcgcttgag acgaagcttt gacaggatat attggcgggt
     1681 aaacctaaga gaaaagagcg tttattagaa taatcggata tttaaaaggg cgtgaaaagg
     1741 tttatccgtt cgtccatttg tatgtgcatg ccaaccacag ggttcccctc gggatcaaag
     1801 tactttgatc caacccctcc gctgctatag tgcagtcggc ttctgacgtt cagtgcagcc
     1861 gtcatctgaa aacgacatgt cgcacaagtc ctaagttacg cgacaggctg ccgccctgcc
     1921 cttttcctgg cgttttcttg tcgcgtgttt tagtcgcata aagtagaata cttgcgacta
     1981 gaaccggaga cattacgcca tgaacaagag cgccgccgct ggcctgctgg gctatgcccg
     2041 cgtcagcacc gacgaccagg acttgaccaa ccaacgggcc gaactgcacg cggccggctg
     2101 caccaagctg ttttccgaga agatcaccgg caccaggcgc gaccgcccgg agctggccag
     2161 gatgcttgac cacctacgcc ctggcgacgt tgtgacagtg accaggctag accgcctggc
     2221 ccgcagcacc cgcgacctac tggacattgc cgagcgcatc caggaggccg gcgcgggcct
     2281 gcgtagcctg gcagagccgt gggccgacac caccacgccg gccggccgca tggtgttgac
     2341 cgtgttcgcc ggcattgccg agttcgagcg ttccctaatc atcgaccgca cccggagcgg
     2401 gcgcgaggcc gccaaggccc gaggcgtgaa gtttggcccc cgccctaccc tcaccccggc
     2461 acagatcgcg cacgcccgcg agctgatcga ccaggaaggc cgcaccgtga aagaggcggc
     2521 tgcactgctt ggcgtgcatc gctcgaccct gtaccgcgca cttgagcgca gcgaggaagt
     2581 gacgcccacc gaggccaggc ggcgcggtgc cttccgtgag gacgcattga ccgaggccga
     2641 cgccctggcg gccgccgaga atgaacgcca agaggaacaa gcatgaaacc gcaccaggac
     2701 ggccaggacg aaccgttttt cattaccgaa gagatcgagg cggagatgat cgcggccggg
     2761 tacgtgttcg agccgcccgc gcacctctca accgtgcggc tgcatgaaat cctggccggt
     2821 ttgtctgatg ccaagctggc ggcctggccg gccagcttgg ccgctgaaga aaccgagcgc
     2881 cgccgtctaa aaaggtgatg tgtatttgag taaaacagct tgcgtcatgc ggtcgctgcg
     2941 tatatgatcc gatgagtaaa taaacaaata cgcaagggga acgcatgaag gttatcgctg
     3001 tacttaacca gaaaggcggg tcaggcaaga cgaccatcgg aacccatcta gcccgcgccc
     3061 tgcaactcgc cggggccgat gttctgttag tcgattccga tccccagggc agtgcccgcg
     3121 attgggcggc cgtgcgggaa gatcaaccgc taaccgttgt cggcatcgac cgcccgacga
     3181 ttgaccgcga cgtgaaggcc atcggccggc gcgacttcgt agtgatcgac ggagcgcccc
     3241 aggcggcgga cttggctgtg tccgcgatca aggcagccga cttcgtgctg attccggtgc
     3301 agccaagccc ttacgacata tgggccaccg ccgacctggt ggagctggtt aagcagcgca
     3361 ttgaggtcac ggatggaagg ctacaagcgg cctttgtcgt gtcgcgggcg atcaaaggca
     3421 cgcgcatcgg cggtgaggtt gccgaggcgc tggccgggta cgagctgccc attcttgagt
     3481 cccgtatcac gcagcgcgtg agctacccag gcactgccgc cgccggcaca accgttcttg
     3541 aatcagaacc cgagggcgac gctgcccgcg aggtccaggc gctggccgct gaaattaaat
     3601 caaaactcat ttgagttaat gaggtaaaga gaaaatgagc aaaagcacaa acacgctaag
     3661 tgccggccgt ccgagcgcac gcagcagcaa ggctgcaacg ttggccagcc tggcagacac
     3721 gccagccatg aagcgggtca actttcagtt gccggcggag gatcacacca agctgaagat
     3781 gtacgcggta cgccaaggca agaccattac cgagctgcta tctgaataga tcgcgcagct
     3841 accagagtaa atgagcaaat gaataaatga gtagatgaat tttagcggct aaaggaggcg
     3901 gcatggaaaa tcaagaacaa ccaggcaccg acgccgtgga atgccccatg tgtggaggaa
     3961 cgggcggttg gccaggcgta agcggctggg ttgtctgccg gccctgcaat ggcactggaa
     4021 cccccaagcc cgaggaatcg gcgtgacggt cgcaaaccat ccggcccggt acaaatcggc
     4081 gcggcgctgg gtgatgacct ggtggagaag ttgaaggccg cgcaggccgc ccagcggcaa
     4141 cgcatcgagg cagaagcacg ccccggtgaa tcgtggcaag cggccgctga tcgaatccgc
     4201 aaagaatccc ggcaaccgcc ggcagccggt gcgccgtcga ttaggaagcc gcccaagggc
     4261 gacgagcaac cagatttttt cgttccgatg ctctatgacg tgggcacccg cgatagtcgc
     4321 agcatcatgg acgtggccgt tttccgtctg tcgaagcgtg accgacgagc tggcgaggtg
     4381 atccgctacg agcttccaga cgggcacgta gaggtttccg cagggccggc cggcatggcc
     4441 agtgtgtggg attacgacct ggtactgatg gcggtttccc atctaaccga atccatgaac
     4501 cgataccggg aagggaaggg agacaagccc ggccgcgtgt tccgtccaca cgttgcggac
     4561 gtactcaagt tctgccggcg agccgatggc ggaaagcaga aagacgacct ggtagaaacc
     4621 tgcattcggt taaacaccac gcacgttgcc atgcagcgta cgaagaaggc caagaacggc
     4681 cgcctggtga cggtatccga gggtgaagcc ttgattagcc gctacaagat cgtaaagagc
     4741 gaaaccgggc ggccggagta catcgagatc gagctagctg attggatgta ccgcgagatc
     4801 acagaaggca agaacccgga cgtgctgacg gttcaccccg attacttttt gatcgatccc
     4861 ggcatcggcc gttttctcta ccgcctggca cgccgcgccg caggcaaggc agaagccaga
     4921 tggttgttca agacgatcta cgaacgcagt ggcagcgccg gagagttcaa gaagttctgt
     4981 ttcaccgtgc gcaagctgat cgggtcaaat gacctgccgg agtacgattt gaaggaggag
     5041 gcggggcagg ctggcccgat cctagtcatg cgctaccgca acctgatcga gggcgaagca
     5101 tccgccggtt cctaatgtac ggagcagatg ctagggcaaa ttgccctagc aggggaaaaa
     5161 ggtcgaaaag gactctttcc tgtggatagc acgtacattg ggaacccaaa gccgtacatt
     5221 gggaaccgga acccgtacat tgggaaccca aagccgtaca ttgggaaccg gtcacacatg
     5281 taagtgactg atataaaaga gaaaaaaggc gatttttccg cctaaaactc tttaaaactt
     5341 attaaaactc ttaaaacccg cctggcctgt gcataactgt ctggccagcg cacagccgaa
     5401 gagctgcaaa aagcgcctac ccttcggtcg ctgcgctccc tacgccccgc cgcttcgcgt
     5461 cggcctatcg cggccgctgg ccgctcaaaa atggctggcc tacggccagg caatctacca
     5521 gggcgcggac aagccgcgcc gtcgccactc gaccgccggc gcccacatca aggcaccctg
     5581 cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggtgacggt
     5641 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     5701 tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtatac
     5761 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
     5821 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc
     5881 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5941 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     6001 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6061 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6121 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6181 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     6241 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6301 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6361 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6421 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6481 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6541 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6601 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6661 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgca ttctaggtga
     6721 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat
     6781 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
     6841 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
     6901 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
     6961 tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca
     7021 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
     7081 cgcctgagcg agtcgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga
     7141 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
     7201 ttcttctaat acctggaatg ctgttttccc tgggatcgca gtggtgagta accatgcatc
     7261 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
     7321 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
     7381 caactctggc gcatcgggct tcccatacaa tcggtagatt gtcgcacctg attgcccgac
//
