LOCUS       sequence-299490-        7448 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 7448)
  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 7448)
  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: pEPYC-alpha-2rKN0009
FEATURES             Location/Qualifiers
     source          1..7448
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1226..1855)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIGTHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    3156..3180
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3403..3915
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             3976..3999
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      4015..4267
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    4295..4319
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             4752..5567
                     /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             4752..5567
                     /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     complement(4825..4844)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5655..6243
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6144..6163
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6397..6414
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6429..6569
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6495..6514)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6655..6677
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      6913..7107
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(join(7173..7448,1..792))
                     /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 ggggtgaacc gtcagcacgt ccgggttctt gccttctgtg atctcgcggt acatccaatc
       61 agctagctcg atctcgatgt actccggccg cccggtttcg ctctttacga tcttgtagcg
      121 gctaatcaag gcttcaccct cggataccgt caccaggcgg ccgttcttgg ccttcttcgt
      181 acgctgcatg gcaacgtgcg tggtgtttaa ccgaatgcag gtttctacca ggtcgtcttt
      241 ctgctttccg ccatcggctc gccggcagaa cttgagtacg tccgcaacgt gtggacggaa
      301 cacgcggccg ggcttgtctc ccttcccttc ccggtatcgg ttcatggatt cggttagatg
      361 ggaaaccgcc atcagtacca ggtcgtaatc ccacacactg gccatgccgg ccggccctgc
      421 ggaaacctct acgtgcccgt ctggaagctc gtagcggatc acctcgccag ctcgtcggtc
      481 acgcttcgac agacggaaaa cggccacgtc catgatgctg cgactatcgc gggtgcccac
      541 gtcatagagc atcggaacga aaaaatctgg ttgctcgtcg cccttgggcg gcttcctaat
      601 cgacggcgca ccggctgccg gcggttgccg ggattctttg cggattcgat cagcggccgc
      661 ttgccacgat tcaccggggc gtgcttctgc ctcgatgcgt tgccgctggg cggcctgcgc
      721 ggccttcaac ttctccacca ggtcatcacc cagcgccgcg ccgatttgta ccgggccgga
      781 tggtttgcga ccgtcacgcc gattcctcgg gcttgggggt tccagtgcca ttgcagggcc
      841 ggcagacaac ccagccgctt acgcctggcc aaccgcccgt tcctccacac atggggcatt
      901 ccacggcgtc ggtgcctggt tgttcttgat tttccatgcc gcctccttta gccgctaaaa
      961 ttcatctact catttattca tttgctcatt tactctggta gctgcgcgat ctattcagat
     1021 agcagctcgg taatggtctt gccttggcgt accgcgtaca tcttcagctt ggtgtgatcc
     1081 tccgccggca actgaaagtt gacccgcttc atggctggcg tgtctgccag gctggccaac
     1141 gttgcagcct tgctgctgcg tgcgctcgga cggccggcac ttagcgtgtt tgtgcttttg
     1201 ctcattttct ctttacctca ttaactcaaa tgagttttga tttaatttca gcggccagcg
     1261 cctggacctc gcgggcagcg tcgccctcgg gttctgattc aagaacggtt gtgccggcgg
     1321 cggcagtgcc tgggtagctc acgcgctgcg tgatacggga ctcaagaatg ggcagctcgt
     1381 acccggccag cgcctcggca acctcaccgc cgatgcgcgt gcctttgatc gcccgcgaca
     1441 cgacaaaggc cgcttgtagc cttccatccg tgacctcaat gcgctgctta accagctcca
     1501 ccaggtcggc ggtggcccat atgtcgtaag ggcttggctg caccggaatc agcacgaagt
     1561 cggctgcctt gatcgcggac acagccaagt ccgccgcctg gggcgctccg tcgatcacta
     1621 cgaagtcgcg ccggccgatg gccttcacgt cgcggtcaat cgtcgggcgg tcgatgccga
     1681 caacggttag cggttgatct tcccgcacgg ccgcccaatc gcgggcactg ccctggggat
     1741 cggaatcgac taacagaaca tcggccccgg cgagttgcag ggcgcgggct agatgggttc
     1801 cgatggtcgt cttgcctgac ccgcctttct ggttaagtac agcgataacc ttcatgcgtt
     1861 ccccttgcgt atttgtttat ttactcatcg gatcatatac gcagcgaccg catgacgcaa
     1921 gctgttttac tcaaatacac atcacctttt tagacggcgg cgctcggttt cttcagcggc
     1981 caagctggcc ggccaggccg ccagcttggc atcagacaaa ccggccagga tttcatgcag
     2041 ccgcacggtt gagaggtgcg cgggcggctc gaacacgtac ccggccgcga tcatctccgc
     2101 ctcgatctct tcggtaatga aaaacggttc gtcctggccg tcctggtgcg gtttcatgct
     2161 tgttcctctt ggcgttcatt ctcggcggcc gccagggcgt cggcctcggt caatgcgtcc
     2221 tcacggaagg caccgcgccg cctggcctcg gtgggcgtca cttcctcgct gcgctcaagt
     2281 gcgcggtaca gggtcgagcg atgcacgcca agcagtgcag ccgcctcttt cacggtgcgg
     2341 ccttcctggt cgatcagctc gcgggcgtgc gcgatctgtg ccggggtgag ggtagggcgg
     2401 gggccaaact tcacgcctcg ggccttggcg gcctcgcgcc cgctccgggt gcggtcgatg
     2461 attagggaac gctcgaactc ggcaatgccg gcgaacacgg tcaacaccat gcggccggcc
     2521 ggcgtggtgg tgtcggccca cggctctgcc aggctacgca ggcccgcgcc ggcctcctgg
     2581 atgcgctcgg caatgtccag taggtcgcgg gtgctgcggg ccaggcggtc tagcctggtc
     2641 actgtcacaa cgtcgccagg gcgtaggtgg tcaagcatcc tggccagctc cgggcggtcg
     2701 cgcctggtgc cggtgatctt ctcggaaaac agcttggtgc agccggccgc gtgcagttcg
     2761 gcccgttggt tggtcaagtc ctggtcgtcg gtgctgacgc gggcatagcc cagcaggcca
     2821 gcggcggcgc tcttgttcat ggcgtaatgt ctccggttct agtcgcaagt attctacttt
     2881 atgcgactaa aacacgcgac aagaaaacgc caggaaaagg gcagggcggc agcctgtcgc
     2941 gtaacttagg acttgtgcga catgtcgttt tcagatgacg gctgcactga acgtcagaag
     3001 ccgactgcac tatagcagcg gaggggttgg atcaaagtac tttgatcccg aggggaaccc
     3061 tgtggttggc atgcacatac aaatggacga acggataaac cttttcacgc ccttttaaat
     3121 atccgattat tctaataaac gctcttttct cttaggttta cccgccaata tatcctgtca
     3181 gatatcgtct caagcggagg ttatcacact tggtaattta gaagaagaaa cccataggtt
     3241 ctgtgggttt tttgggtaga agaagaaacc cataggttct gtgggttttt tgggtagaag
     3301 aagaaaccca taggttctgt gggttttttg ggtagaagaa gaaacccata gctatataat
     3361 gagaaacagg ggcaaatccc cgaaaacaaa taacgcgaag caatggtatt cacactcgaa
     3421 gatttcgttg gggactggcg acagacagcc ggctacaacc tggaccaagt ccttgaacag
     3481 ggaggtgtgt ccagtttgtt tcagaatctc ggggtgtccg taactccgat ccaaaggatt
     3541 gtcctgagcg gtgaaaatgg gctgaagatc gacatccatg tcatcatccc gtatgaaggt
     3601 ctgagcggcg accaaatggg ccagatcgaa aaaattttta aggtggtgta ccctgtggat
     3661 gatcatcact ttaaggtgat cctgcactat ggcacactgg taatcgacgg ggttacgccg
     3721 aacatgatcg actatttcgg acggccgtat gaaggcatcg ccgtgttcga cggcaaaaag
     3781 atcactgtaa cagggaccct gtggaacggc aacaaaatta tcgacgagcg cctgatcaac
     3841 cccgacggct ccctgctgtt ccgagtaacc atcaacggag tgaccggctg gcggctgtgc
     3901 gaacgcattc tggcttcgga ttataaggac catgacggag actataagga ccatgacctc
     3961 gacgctgcag cagcggatta taaggacgat gacgataagt gagcttgtca agcagatcgt
     4021 tcaaacattt ggcaataaag tttcttaaga ttgaatcctg ttgccggtct tgcgatgatt
     4081 atcatataat ttctgttgaa ttacgttaag catgtaataa ttaacatgta atgcatgacg
     4141 ttatttatga gatgggtttt tatgattaga gtcccgcaat tatacattta atacgcgata
     4201 gaaaacaaaa tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc atctatgtta
     4261 ctagatcgac gcttgactga gacgatatcg ttatgtttac accacaatat atcctgccac
     4321 cagccagcca acagctcccc gaccggcagc tcggcacaaa atcaccactc gatacaggca
     4381 gcccatcagt ccgggacggc gtcagcggga gagccgttgt aaggcggcag actttgctca
     4441 tgttaccgat gctattcgga agaacggcaa ctaagctgcc gggtttgaaa cacggatgat
     4501 ctcgcggagg gtagcatgtt gattgtaacg atgacagagc gttgctgcct gtgatcaccg
     4561 cggtttcaaa atcggctccg tcgatactat gttatacgcc aactttgaaa acaactttga
     4621 aaaagctgtt ttctggtatt taaggtttta gaatgcaagg aacagtgaat tggagttcgt
     4681 cttgttataa ttagcttctt ggggtatctt taaatactgt agaaaagagg aaggaaataa
     4741 taagttctgt tatgagccat attcaacggg aaacgtcttg ctcaaggccg cgattaaatt
     4801 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag
     4861 gtgcgacaat ctaccgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg
     4921 gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg
     4981 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac
     5041 tcaccactgc gatcccaggg aaaacagcat tccaggtatt agaagaatat cctgattcag
     5101 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt
     5161 gtaattgtcc ttttaacagc gatcgcgtat ttcgactcgc tcaggcgcaa tcacgaatga
     5221 ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac
     5281 aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg
     5341 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg
     5401 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg
     5461 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg
     5521 atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca cctagaatgc
     5581 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5641 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5701 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5761 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     5821 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5881 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5941 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6001 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6061 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6121 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6181 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6241 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6301 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     6361 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     6421 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     6481 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
     6541 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     6601 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtcaccggga
     6661 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ggtgccttga
     6721 tgtgggcgcc ggcggtcgag tggcgacggc gcggcttgtc cgcgccctgg tagattgcct
     6781 ggccgtaggc cagccatttt tgagcggcca gcggccgcga taggccgacg cgaagcggcg
     6841 gggcgtaggg agcgcagcga ccgaagggta ggcgcttttt gcagctcttc ggctgtgcgc
     6901 tggccagaca gttatgcaca ggccaggcgg gttttaagag ttttaataag ttttaaagag
     6961 ttttaggcgg aaaaatcgcc ttttttctct tttatatcag tcacttacat gtgtgaccgg
     7021 ttcccaatgt acggctttgg gttcccaatg tacgggttcc ggttcccaat gtacggcttt
     7081 gggttcccaa tgtacgtgct atccacagga aagagtcctt ttcgaccttt ttcccctgct
     7141 agggcaattt gccctagcat ctgctccgta cattaggaac cggcggatgc ttcgccctcg
     7201 atcaggttgc ggtagcgcat gactaggatc gggccagcct gccccgcctc ctccttcaaa
     7261 tcgtactccg gcaggtcatt tgacccgatc agcttgcgca cggtgaaaca gaacttcttg
     7321 aactctccgg cgctgccact gcgttcgtag atcgtcttga acaaccatct ggcttctgcc
     7381 ttgcctgcgg cgcggcgtgc caggcggtag agaaaacggc cgatgccggg atcgatcaaa
     7441 aagtaatc
//
