LOCUS       sequence-299489-        8538 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 8538)
  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 8538)
  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-1KN0008
FEATURES             Location/Qualifiers
     source          1..8538
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      154..348
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(584..606)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    692..832
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     747..766
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(847..864)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1018..1606)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1098..1117)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1694..2509)
                     /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(1694..2509)
                     /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     2417..2436
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    2942..2966
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             3191..4840
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(3257..3275)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     CDS             4904..4927
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      4950..5150
                     /label=gene 7 terminator
                     /note="transcription terminator for octopine-type Ti 
                     plasmid gene 7"
     misc_feature    5171..5195
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             6496..7125
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIGTHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             join(7559..8538,1..88)
                     /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 tttgaaggag gaggcggggc aggctggccc gatcctagtc atgcgctacc gcaacctgat
       61 cgagggcgaa gcatccgccg gttcctaatg tacggagcag atgctagggc aaattgccct
      121 agcaggggaa aaaggtcgaa aaggactctt tcctgtggat agcacgtaca ttgggaaccc
      181 aaagccgtac attgggaacc ggaacccgta cattgggaac ccaaagccgt acattgggaa
      241 ccggtcacac atgtaagtga ctgatataaa agagaaaaaa ggcgattttt ccgcctaaaa
      301 ctctttaaaa cttattaaaa ctcttaaaac ccgcctggcc tgtgcataac tgtctggcca
      361 gcgcacagcc gaagagctgc aaaaagcgcc tacccttcgg tcgctgcgct ccctacgccc
      421 cgccgcttcg cgtcggccta tcgcggccgc tggccgctca aaaatggctg gcctacggcc
      481 aggcaatcta ccagggcgcg gacaagccgc gccgtcgcca ctcgaccgcc ggcgcccaca
      541 tcaaggcacc ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
      601 tcccggtgac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
      661 gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata
      721 gcggagtgta tactggctta actatgcggc atcagagcag attgtactga gagtgcacca
      781 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc
      841 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
      901 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
      961 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1021 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1081 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1141 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1201 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1261 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     1321 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     1381 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     1441 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     1501 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     1561 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     1621 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     1681 gcattctagg tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat
     1741 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac
     1801 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa
     1861 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac
     1921 catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt
     1981 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat
     2041 tcattcgtga ttgcgcctga gcgagtcgaa atacgcgatc gctgttaaaa ggacaattac
     2101 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
     2161 ctgaatcagg atattcttct aatacctgga atgctgtttt ccctgggatc gcagtggtga
     2221 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
     2281 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc
     2341 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcggtag attgtcgcac
     2401 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
     2461 aatttaatcg cggccttgag caagacgttt cccgttgaat atggctcata acagaactta
     2521 ttatttcctt cctcttttct acagtattta aagatacccc aagaagctaa ttataacaag
     2581 acgaactcca attcactgtt ccttgcattc taaaacctta aataccagaa aacagctttt
     2641 tcaaagttgt tttcaaagtt ggcgtataac atagtatcga cggagccgat tttgaaaccg
     2701 cggtgatcac aggcagcaac gctctgtcat cgttacaatc aacatgctac cctccgcgag
     2761 atcatccgtg tttcaaaccc ggcagcttag ttgccgttct tccgaatagc atcggtaaca
     2821 tgagcaaagt ctgccgcctt acaacggctc tcccgctgac gccgtcccgg actgatgggc
     2881 tgcctgtatc gagtggtgat tttgtgccga gctgccggtc ggggagctgt tggctggctg
     2941 gtggcaggat atattgtggt gtaaacataa cgaattcgtc tcaggaggtt atcacacttg
     3001 gtaatttaga agaagaaacc cataggttct gtgggttttt tgggtagaag aagaaaccca
     3061 taggttctgt gggttttttg ggtagaagaa gaaacccata ggttctgtgg gttttttggg
     3121 tagaagaaga aacccatagc tatataatga gaaacagggg caaatccccg aaaacaaata
     3181 acgcgaagca atggaggacg ccaaaaacat aaagaaaggc ccggcgccat tctatccgct
     3241 ggaagatgga accgctggag agcaactgca taaggctatg aagagatacg ccctggttcc
     3301 tggaacaatt gcttttacag atgcacatat cgaggtggac atcacttacg ctgagtactt
     3361 cgaaatgtcc gttcggttgg cagaagctat gaaacgatat gggctgaata caaatcacag
     3421 aatcgtcgta tgcagtgaaa actctcttca attctttatg ccggtgttgg gcgcgttatt
     3481 tatcggagtt gcagttgcgc ccgcgaacga catttataat gaacgtgaat tgctcaacag
     3541 tatgggcatt tcgcagccta ccgtggtgtt cgtttccaaa aaggggttgc aaaaaatttt
     3601 gaacgtgcaa aaaaagctcc caatcatcca aaaaattatt atcatggatt ctaaaacgga
     3661 ttaccaggga tttcagtcga tgtacacgtt cgtcacatct catctacctc ccggttttaa
     3721 tgaatacgat tttgtgccag agtccttcga tagggacaag acaattgcac tgatcatgaa
     3781 ctcctctgga tctactggtc tgcctaaagg tgtcgctctg cctcatagaa ctgcctgcgt
     3841 gagattctcg catgccagag atcctatttt tggcaatcaa atcattccgg atactgcgat
     3901 tttaagtgtt gttccattcc atcacggttt tggaatgttt actacactcg gatatttgat
     3961 atgtggattt cgagtcgtct taatgtatag atttgaagaa gaactgtttc tgaggagcct
     4021 tcaggattac aagattcaaa gtgcgctgct ggtgccaacc ctattctcct tcttcgccaa
     4081 aagcactctg attgacaaat acgatttatc taatttacac gaaattgctt ctggtggcgc
     4141 tcccctctct aaggaagtcg gggaagcggt tgccaagagg ttccatctgc caggtatcag
     4201 gcaaggatat gggctcactg agactacatc agctattctg attacacccg agggggatga
     4261 taaaccgggc gcggtcggta aagttgttcc attttttgaa gcgaaggttg tggatctgga
     4321 taccgggaaa acgctgggcg ttaatcaaag aggcgaactg tgtgtgagag gtcctatgat
     4381 tatgtccggt tatgtaaaca atccggaagc gaccaacgcc ttgattgaca aggatggatg
     4441 gctacattct ggagacatag cttactggga cgaggacgaa cacttcttca tcgttgaccg
     4501 cctgaagtct ctgattaagt acaaaggcta tcaggtggct cccgctgaat tggaatccat
     4561 cttgctccaa caccccaaca tcttcgacgc aggtgtcgca ggtctgcccg acgatgacgc
     4621 cggtgaactt cccgccgccg ttgttgtttt ggagcacgga aagacgatga cggaaaaaga
     4681 gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt
     4741 gtttgtggac gaagtaccga aaggtcttac cggaaaactc gacgcaagaa aaatcagaga
     4801 gatcctcata aaggccaaga agggcggaaa gatcgccgtg ggttcggatt ataaggacca
     4861 tgacggagac tataaggacc atgacctcga cgctgcagca gcggattata aggacgatga
     4921 cgataagtga gcttgatccc ccgtcgacag ctagctatat catcaattta tgtattacac
     4981 ataatatcgc actcagtctt tcatctacgg caatgtacca gctgatataa tcagttattg
     5041 aaatatttct gaatttaaac ttgcatcaat aaatttatgt ttttgcttgg actataatac
     5101 ctgacttgtt attttatcaa taaatattta aactatattt ctttcaagat cgctgtcatg
     5161 agacgaattc tgacaggata tattggcggg taaacctaag agaaaagagc gtttattaga
     5221 ataatcggat atttaaaagg gcgtgaaaag gtttatccgt tcgtccattt gtatgtgcat
     5281 gccaaccaca gggttcccct cgggatcaaa gtactttgat ccaacccctc cgctgctata
     5341 gtgcagtcgg cttctgacgt tcagtgcagc cgtcatctga aaacgacatg tcgcacaagt
     5401 cctaagttac gcgacaggct gccgccctgc ccttttcctg gcgttttctt gtcgcgtgtt
     5461 ttagtcgcat aaagtagaat acttgcgact agaaccggag acattacgcc atgaacaaga
     5521 gcgccgccgc tggcctgctg ggctatgccc gcgtcagcac cgacgaccag gacttgacca
     5581 accaacgggc cgaactgcac gcggccggct gcaccaagct gttttccgag aagatcaccg
     5641 gcaccaggcg cgaccgcccg gagctggcca ggatgcttga ccacctacgc cctggcgacg
     5701 ttgtgacagt gaccaggcta gaccgcctgg cccgcagcac ccgcgaccta ctggacattg
     5761 ccgagcgcat ccaggaggcc ggcgcgggcc tgcgtagcct ggcagagccg tgggccgaca
     5821 ccaccacgcc ggccggccgc atggtgttga ccgtgttcgc cggcattgcc gagttcgagc
     5881 gttccctaat catcgaccgc acccggagcg ggcgcgaggc cgccaaggcc cgaggcgtga
     5941 agtttggccc ccgccctacc ctcaccccgg cacagatcgc gcacgcccgc gagctgatcg
     6001 accaggaagg ccgcaccgtg aaagaggcgg ctgcactgct tggcgtgcat cgctcgaccc
     6061 tgtaccgcgc acttgagcgc agcgaggaag tgacgcccac cgaggccagg cggcgcggtg
     6121 ccttccgtga ggacgcattg accgaggccg acgccctggc ggccgccgag aatgaacgcc
     6181 aagaggaaca agcatgaaac cgcaccagga cggccaggac gaaccgtttt tcattaccga
     6241 agagatcgag gcggagatga tcgcggccgg gtacgtgttc gagccgcccg cgcacctctc
     6301 aaccgtgcgg ctgcatgaaa tcctggccgg tttgtctgat gccaagctgg cggcctggcc
     6361 ggccagcttg gccgctgaag aaaccgagcg ccgccgtcta aaaaggtgat gtgtatttga
     6421 gtaaaacagc ttgcgtcatg cggtcgctgc gtatatgatc cgatgagtaa ataaacaaat
     6481 acgcaagggg aacgcatgaa ggttatcgct gtacttaacc agaaaggcgg gtcaggcaag
     6541 acgaccatcg gaacccatct agcccgcgcc ctgcaactcg ccggggccga tgttctgtta
     6601 gtcgattccg atccccaggg cagtgcccgc gattgggcgg ccgtgcggga agatcaaccg
     6661 ctaaccgttg tcggcatcga ccgcccgacg attgaccgcg acgtgaaggc catcggccgg
     6721 cgcgacttcg tagtgatcga cggagcgccc caggcggcgg acttggctgt gtccgcgatc
     6781 aaggcagccg acttcgtgct gattccggtg cagccaagcc cttacgacat atgggccacc
     6841 gccgacctgg tggagctggt taagcagcgc attgaggtca cggatggaag gctacaagcg
     6901 gcctttgtcg tgtcgcgggc gatcaaaggc acgcgcatcg gcggtgaggt tgccgaggcg
     6961 ctggccgggt acgagctgcc cattcttgag tcccgtatca cgcagcgcgt gagctaccca
     7021 ggcactgccg ccgccggcac aaccgttctt gaatcagaac ccgagggcga cgctgcccgc
     7081 gaggtccagg cgctggccgc tgaaattaaa tcaaaactca tttgagttaa tgaggtaaag
     7141 agaaaatgag caaaagcaca aacacgctaa gtgccggccg tccgagcgca cgcagcagca
     7201 aggctgcaac gttggccagc ctggcagaca cgccagccat gaagcgggtc aactttcagt
     7261 tgccggcgga ggatcacacc aagctgaaga tgtacgcggt acgccaaggc aagaccatta
     7321 ccgagctgct atctgaatag atcgcgcagc taccagagta aatgagcaaa tgaataaatg
     7381 agtagatgaa ttttagcggc taaaggaggc ggcatggaaa atcaagaaca accaggcacc
     7441 gacgccgtgg aatgccccat gtgtggagga acgggcggtt ggccaggcgt aagcggctgg
     7501 gttgtctgcc ggccctgcaa tggcactgga acccccaagc ccgaggaatc ggcgtgacgg
     7561 tcgcaaacca tccggcccgg tacaaatcgg cgcggcgctg ggtgatgacc tggtggagaa
     7621 gttgaaggcc gcgcaggccg cccagcggca acgcatcgag gcagaagcac gccccggtga
     7681 atcgtggcaa gcggccgctg atcgaatccg caaagaatcc cggcaaccgc cggcagccgg
     7741 tgcgccgtcg attaggaagc cgcccaaggg cgacgagcaa ccagattttt tcgttccgat
     7801 gctctatgac gtgggcaccc gcgatagtcg cagcatcatg gacgtggccg ttttccgtct
     7861 gtcgaagcgt gaccgacgag ctggcgaggt gatccgctac gagcttccag acgggcacgt
     7921 agaggtttcc gcagggccgg ccggcatggc cagtgtgtgg gattacgacc tggtactgat
     7981 ggcggtttcc catctaaccg aatccatgaa ccgataccgg gaagggaagg gagacaagcc
     8041 cggccgcgtg ttccgtccac acgttgcgga cgtactcaag ttctgccggc gagccgatgg
     8101 cggaaagcag aaagacgacc tggtagaaac ctgcattcgg ttaaacacca cgcacgttgc
     8161 catgcagcgt acgaagaagg ccaagaacgg ccgcctggtg acggtatccg agggtgaagc
     8221 cttgattagc cgctacaaga tcgtaaagag cgaaaccggg cggccggagt acatcgagat
     8281 cgagctagct gattggatgt accgcgagat cacagaaggc aagaacccgg acgtgctgac
     8341 ggttcacccc gattactttt tgatcgatcc cggcatcggc cgttttctct accgcctggc
     8401 acgccgcgcc gcaggcaagg cagaagccag atggttgttc aagacgatct acgaacgcag
     8461 tggcagcgcc ggagagttca agaagttctg tttcaccgtg cgcaagctga tcgggtcaaa
     8521 tgacctgccg gagtacga
//
