LOCUS       sequence-299491-        7334 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 7334)
  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 7334)
  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-2rKN0010
FEATURES             Location/Qualifiers
     source          1..7334
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1220..1849)
                     /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    3150..3174
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3283..3795
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             3856..3879
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      3895..4147
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    4175..4199
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             4632..5447
                     /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             4632..5447
                     /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(4705..4724)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5535..6123
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6024..6043
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6277..6294
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6309..6449
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6375..6394)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6535..6557
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      6793..6987
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(join(7053..7334,1..786))
                     /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 aaccgtcagc acgtccgggt tcttgccttc tgtgatctcg cggtacatcc aatcagctag
       61 ctcgatctcg atgtactccg gccgcccggt ttcgctcttt acgatcttgt agcggctaat
      121 caaggcttca ccctcggata ccgtcaccag gcggccgttc ttggccttct tcgtacgctg
      181 catggcaacg tgcgtggtgt ttaaccgaat gcaggtttct accaggtcgt ctttctgctt
      241 tccgccatcg gctcgccggc agaacttgag tacgtccgca acgtgtggac ggaacacgcg
      301 gccgggcttg tctcccttcc cttcccggta tcggttcatg gattcggtta gatgggaaac
      361 cgccatcagt accaggtcgt aatcccacac actggccatg ccggccggcc ctgcggaaac
      421 ctctacgtgc ccgtctggaa gctcgtagcg gatcacctcg ccagctcgtc ggtcacgctt
      481 cgacagacgg aaaacggcca cgtccatgat gctgcgacta tcgcgggtgc ccacgtcata
      541 gagcatcgga acgaaaaaat ctggttgctc gtcgcccttg ggcggcttcc taatcgacgg
      601 cgcaccggct gccggcggtt gccgggattc tttgcggatt cgatcagcgg ccgcttgcca
      661 cgattcaccg gggcgtgctt ctgcctcgat gcgttgccgc tgggcggcct gcgcggcctt
      721 caacttctcc accaggtcat cacccagcgc cgcgccgatt tgtaccgggc cggatggttt
      781 gcgaccgtca cgccgattcc tcgggcttgg gggttccagt gccattgcag ggccggcaga
      841 caacccagcc gcttacgcct ggccaaccgc ccgttcctcc acacatgggg cattccacgg
      901 cgtcggtgcc tggttgttct tgattttcca tgccgcctcc tttagccgct aaaattcatc
      961 tactcattta ttcatttgct catttactct ggtagctgcg cgatctattc agatagcagc
     1021 tcggtaatgg tcttgccttg gcgtaccgcg tacatcttca gcttggtgtg atcctccgcc
     1081 ggcaactgaa agttgacccg cttcatggct ggcgtgtctg ccaggctggc caacgttgca
     1141 gccttgctgc tgcgtgcgct cggacggccg gcacttagcg tgtttgtgct tttgctcatt
     1201 ttctctttac ctcattaact caaatgagtt ttgatttaat ttcagcggcc agcgcctgga
     1261 cctcgcgggc agcgtcgccc tcgggttctg attcaagaac ggttgtgccg gcggcggcag
     1321 tgcctgggta gctcacgcgc tgcgtgatac gggactcaag aatgggcagc tcgtacccgg
     1381 ccagcgcctc ggcaacctca ccgccgatgc gcgtgccttt gatcgcccgc gacacgacaa
     1441 aggccgcttg tagccttcca tccgtgacct caatgcgctg cttaaccagc tccaccaggt
     1501 cggcggtggc ccatatgtcg taagggcttg gctgcaccgg aatcagcacg aagtcggctg
     1561 ccttgatcgc ggacacagcc aagtccgccg cctggggcgc tccgtcgatc actacgaagt
     1621 cgcgccggcc gatggccttc acgtcgcggt caatcgtcgg gcggtcgatg ccgacaacgg
     1681 ttagcggttg atcttcccgc acggccgccc aatcgcgggc actgccctgg ggatcggaat
     1741 cgactaacag aacatcggcc ccggcgagtt gcagggcgcg ggctagatgg gttccgatgg
     1801 tcgtcttgcc tgacccgcct ttctggttaa gtacagcgat aaccttcatg cgttcccctt
     1861 gcgtatttgt ttatttactc atcggatcat atacgcagcg accgcatgac gcaagctgtt
     1921 ttactcaaat acacatcacc tttttagacg gcggcgctcg gtttcttcag cggccaagct
     1981 ggccggccag gccgccagct tggcatcaga caaaccggcc aggatttcat gcagccgcac
     2041 ggttgagagg tgcgcgggcg gctcgaacac gtacccggcc gcgatcatct ccgcctcgat
     2101 ctcttcggta atgaaaaacg gttcgtcctg gccgtcctgg tgcggtttca tgcttgttcc
     2161 tcttggcgtt cattctcggc ggccgccagg gcgtcggcct cggtcaatgc gtcctcacgg
     2221 aaggcaccgc gccgcctggc ctcggtgggc gtcacttcct cgctgcgctc aagtgcgcgg
     2281 tacagggtcg agcgatgcac gccaagcagt gcagccgcct ctttcacggt gcggccttcc
     2341 tggtcgatca gctcgcgggc gtgcgcgatc tgtgccgggg tgagggtagg gcgggggcca
     2401 aacttcacgc ctcgggcctt ggcggcctcg cgcccgctcc gggtgcggtc gatgattagg
     2461 gaacgctcga actcggcaat gccggcgaac acggtcaaca ccatgcggcc ggccggcgtg
     2521 gtggtgtcgg cccacggctc tgccaggcta cgcaggcccg cgccggcctc ctggatgcgc
     2581 tcggcaatgt ccagtaggtc gcgggtgctg cgggccaggc ggtctagcct ggtcactgtc
     2641 acaacgtcgc cagggcgtag gtggtcaagc atcctggcca gctccgggcg gtcgcgcctg
     2701 gtgccggtga tcttctcgga aaacagcttg gtgcagccgg ccgcgtgcag ttcggcccgt
     2761 tggttggtca agtcctggtc gtcggtgctg acgcgggcat agcccagcag gccagcggcg
     2821 gcgctcttgt tcatggcgta atgtctccgg ttctagtcgc aagtattcta ctttatgcga
     2881 ctaaaacacg cgacaagaaa acgccaggaa aagggcaggg cggcagcctg tcgcgtaact
     2941 taggacttgt gcgacatgtc gttttcagat gacggctgca ctgaacgtca gaagccgact
     3001 gcactatagc agcggagggg ttggatcaaa gtactttgat cccgagggga accctgtggt
     3061 tggcatgcac atacaaatgg acgaacggat aaaccttttc acgccctttt aaatatccga
     3121 ttattctaat aaacgctctt ttctcttagg tttacccgcc aatatatcct gtcagatatc
     3181 gtctcaagcg gaggttctgt gggttttttg ggtagaagaa gaaacccata gctatataat
     3241 gagaaacagg ggcaaatccc cgaaaacaaa taacgcgaag caatggtatt cacactcgaa
     3301 gatttcgttg gggactggcg acagacagcc ggctacaacc tggaccaagt ccttgaacag
     3361 ggaggtgtgt ccagtttgtt tcagaatctc ggggtgtccg taactccgat ccaaaggatt
     3421 gtcctgagcg gtgaaaatgg gctgaagatc gacatccatg tcatcatccc gtatgaaggt
     3481 ctgagcggcg accaaatggg ccagatcgaa aaaattttta aggtggtgta ccctgtggat
     3541 gatcatcact ttaaggtgat cctgcactat ggcacactgg taatcgacgg ggttacgccg
     3601 aacatgatcg actatttcgg acggccgtat gaaggcatcg ccgtgttcga cggcaaaaag
     3661 atcactgtaa cagggaccct gtggaacggc aacaaaatta tcgacgagcg cctgatcaac
     3721 cccgacggct ccctgctgtt ccgagtaacc atcaacggag tgaccggctg gcggctgtgc
     3781 gaacgcattc tggcttcgga ttataaggac catgacggag actataagga ccatgacctc
     3841 gacgctgcag cagcggatta taaggacgat gacgataagt gagcttgtca agcagatcgt
     3901 tcaaacattt ggcaataaag tttcttaaga ttgaatcctg ttgccggtct tgcgatgatt
     3961 atcatataat ttctgttgaa ttacgttaag catgtaataa ttaacatgta atgcatgacg
     4021 ttatttatga gatgggtttt tatgattaga gtcccgcaat tatacattta atacgcgata
     4081 gaaaacaaaa tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc atctatgtta
     4141 ctagatcgac gcttgactga gacgatatcg ttatgtttac accacaatat atcctgccac
     4201 cagccagcca acagctcccc gaccggcagc tcggcacaaa atcaccactc gatacaggca
     4261 gcccatcagt ccgggacggc gtcagcggga gagccgttgt aaggcggcag actttgctca
     4321 tgttaccgat gctattcgga agaacggcaa ctaagctgcc gggtttgaaa cacggatgat
     4381 ctcgcggagg gtagcatgtt gattgtaacg atgacagagc gttgctgcct gtgatcaccg
     4441 cggtttcaaa atcggctccg tcgatactat gttatacgcc aactttgaaa acaactttga
     4501 aaaagctgtt ttctggtatt taaggtttta gaatgcaagg aacagtgaat tggagttcgt
     4561 cttgttataa ttagcttctt ggggtatctt taaatactgt agaaaagagg aaggaaataa
     4621 taagttctgt tatgagccat attcaacggg aaacgtcttg ctcaaggccg cgattaaatt
     4681 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag
     4741 gtgcgacaat ctaccgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg
     4801 gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg
     4861 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac
     4921 tcaccactgc gatcccaggg aaaacagcat tccaggtatt agaagaatat cctgattcag
     4981 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt
     5041 gtaattgtcc ttttaacagc gatcgcgtat ttcgactcgc tcaggcgcaa tcacgaatga
     5101 ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac
     5161 aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg
     5221 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg
     5281 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg
     5341 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg
     5401 atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca cctagaatgc
     5461 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5521 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5581 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5641 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     5701 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5761 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5821 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5881 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5941 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6001 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6061 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6121 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6181 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     6241 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     6301 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     6361 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
     6421 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     6481 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtcaccggga
     6541 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ggtgccttga
     6601 tgtgggcgcc ggcggtcgag tggcgacggc gcggcttgtc cgcgccctgg tagattgcct
     6661 ggccgtaggc cagccatttt tgagcggcca gcggccgcga taggccgacg cgaagcggcg
     6721 gggcgtaggg agcgcagcga ccgaagggta ggcgcttttt gcagctcttc ggctgtgcgc
     6781 tggccagaca gttatgcaca ggccaggcgg gttttaagag ttttaataag ttttaaagag
     6841 ttttaggcgg aaaaatcgcc ttttttctct tttatatcag tcacttacat gtgtgaccgg
     6901 ttcccaatgt acggctttgg gttcccaatg tacgggttcc ggttcccaat gtacggcttt
     6961 gggttcccaa tgtacgtgct atccacagga aagagtcctt ttcgaccttt ttcccctgct
     7021 agggcaattt gccctagcat ctgctccgta cattaggaac cggcggatgc ttcgccctcg
     7081 atcaggttgc ggtagcgcat gactaggatc gggccagcct gccccgcctc ctccttcaaa
     7141 tcgtactccg gcaggtcatt tgacccgatc agcttgcgca cggtgaaaca gaacttcttg
     7201 aactctccgg cgctgccact gcgttcgtag atcgtcttga acaaccatct ggcttctgcc
     7261 ttgcctgcgg cgcggcgtgc caggcggtag agaaaacggc cgatgccggg atcgatcaaa
     7321 aagtaatcgg ggtg
//
