LOCUS       sequence-182999-        7702 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module for the expression of the Renilla Luciferase with the 
            silencing suppressor P19.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7702)
  AUTHORS   Vazquez-Vilar M, Bernabe-Orts JM, Fernandez-Del-Carmen A, Ziarsolo 
            P, Blanca J, Granell A, Orzaez D
  TITLE     A modular toolbox for gRNA-Cas9 genome engineering in plants based 
            on the GoldenBraid standard.
  JOURNAL   Plant Methods. 2016 Feb 1;12:10. doi: 10.1186/s13007-016-0101-2. 
            eCollection 2016.
  PUBMED    26839579
REFERENCE   2  (bases 1 to 7702)
  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: "Plant 
            Methods."; date: "2016-02-1"; volume: "12:10. doi"; pages: " 
            10.1186/s13007-016-0101-2. eCollection 2016"
COMMENT     Sequence Label: pEGB SF-35S:Renilla:TNOS-35S:P19:TNOS (GB0160)
FEATURES             Location/Qualifiers
     source          1..7702
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        944..1288
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             1362..2297
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     terminator      2324..2576
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        3420..3764
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     terminator      4380..4632
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    4869..4893
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5235..5823)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5994..6809)
                     /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(5994..6809)
                     /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     6717..6736
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6801..6820
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      7100..7535
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    7662..7684
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
ORIGIN
        1 ggagagcgaa atgattgatg aagaacaatg gtggatgaag aacaaagaag gagggagctt
       61 ttgttcaaga tgaacaaaga acaatagtgg atgaagaaca aagtgaaaaa aataaaaaaa
      121 aatgtatggt taaataaaga gtaaagttac cattgagact cctgacagcg aaatgattga
      181 tgaagaacaa tggtggatga agaacaaaga aggagggagc ttttgttcaa gatgaacaaa
      241 gaacaatagt ggatgaagaa caaagtgaaa aaaataaaaa aaaatgtatg gttaaataaa
      301 gagtaaagtt accattgaga ctccgtcagg agactagagc caagctgatc tcctttgccc
      361 cggagatcac catggacgac tttctctatc tctacgatct aggaagaaag ttcgacggag
      421 aaggtgacga taccatgttc accaccgata atgagaagat tagcctcttc aatttcagaa
      481 agaatgctga cccacagatg gttagagagg cctacgcggc aggtctgatc aagacgatct
      541 acccgagtaa taatctccag gagatcaaat accttcccaa gaaggttaaa gatgcagtca
      601 aaagattcag gactaactgc atcaagaaca cagagaaaga tatatttctc aagatcagaa
      661 gtactattcc agtatggacg attcaaggct tgcttcataa accaaggcaa gtaatagaga
      721 ttggagtctc taagaaagta gttcctactg aatcaaaggc catggagtca aaaattcaga
      781 tcgaggatct aacagaactc gccgtgaaga ctggcgaaca gttcatacag agtcttttac
      841 gactcaatga caagaagaaa atcttcgtca acatggtgga gcacgacact ctcgtctact
      901 ccaagaatat caaagataca gtctcagaag accaaagggc tattgagact tttcaacaaa
      961 gggtaatatc gggaaacctc ctcggattcc attgcccagc tatctgtcac ttcatcaaaa
     1021 ggacagtaga aaaggaaggt ggcacctaca aatgccatca ttgcgataaa ggaaaggcta
     1081 tcgttcaaga tgcccctgcc gacagtggtc ccaaagatgg acccccaccc acgaggagca
     1141 tcgtggaaaa agaagacgtt ccaaccacgt cttcaaagca agtggattga tgtgatatct
     1201 ccactgacgt aagggatgac gcacaatccc actatccttc gcaagaccct tcctctatat
     1261 aaggaagttc atttcatttg gagaggactc cggtattttt acaacaatta ccacaacaaa
     1321 acaaacaaca aacaacatta caatttacta ttctagtcga aatgacttcg aaagtttatg
     1381 atccagaaca aaggaaacgg atgataactg gtccgcagtg gtgggccaga tgtaaacaaa
     1441 tgaatgttct tgattcattt attaattatt atgattcaga aaaacatgca gaaaatgctg
     1501 ttattttttt acatggtaac gcggcctctt cttatttatg gcgacatgtt gtgccacata
     1561 ttgagccagt agcgcggtgt attataccag accttattgg tatgggcaaa tcaggcaaat
     1621 ctggtaatgg ttcttatagg ttacttgatc attacaaata tcttactgca tggtttgaac
     1681 ttcttaattt accaaagaag atcatttttg tcggccatga ttggggtgct tgtttggcat
     1741 ttcattatag ctatgagcat caagataaga tcaaagcaat agttcacgct gaaagtgtag
     1801 tagatgtgat tgaatcatgg gatgaatggc ctgatattga agaagatatt gcgttgatca
     1861 aatctgaaga aggagaaaaa atggttttgg agaataactt cttcgtggaa accatgttgc
     1921 catcaaaaat catgagaaag ttagaaccag aagaatttgc agcatatctt gaaccattca
     1981 aagagaaagg tgaagttcgt cgtccaacat tatcatggcc tcgtgaaatc ccgttagtaa
     2041 aaggtggtaa acctgacgtt gtacaaattg ttaggaatta taatgcttat ctacgtgcaa
     2101 gtgatgattt accaaaaatg tttattgaat cggacccagg attcttttcc aatgctattg
     2161 ttgaaggtgc caagaagttt cctaatactg aatttgtcaa agtaaaaggt cttcattttt
     2221 cgcaagaaga tgcacctgat gaaatgggaa aatatatcaa atcgttcgtt gagcgagttc
     2281 tcaaaaatga acaataatga gcttggaatg gatcttcgat cccgatcgtt caaacatttg
     2341 gcaataaagt ttcttaagat tgaatcctgt tgccggtctt gcgacgatta tcatataatt
     2401 tctgttgaat tacgttaagc atgtaataat taacatgtaa tgcatgacgt tatttatgag
     2461 atgggttttt atgattagag tcccgcaatt atacatttaa tacgcgatag aaaacaaaat
     2521 atagcgcgca aactaggata aattatcgcg cgcggtgtca tctatgttac tagatcggga
     2581 attgccaagc taattcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta
     2641 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     2701 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     2761 aaccctgata aatgcttcaa taatgggacc gactcgcgct gtcaggagac tagagccaag
     2821 ctgatctcct ttgccccgga gatcaccatg gacgactttc tctatctcta cgatctagga
     2881 agaaagttcg acggagaagg tgacgatacc atgttcacca ccgataatga gaagattagc
     2941 ctcttcaatt tcagaaagaa tgctgaccca cagatggtta gagaggccta cgcggcaggt
     3001 ctgatcaaga cgatctaccc gagtaataat ctccaggaga tcaaatacct tcccaagaag
     3061 gttaaagatg cagtcaaaag attcaggact aactgcatca agaacacaga gaaagatata
     3121 tttctcaaga tcagaagtac tattccagta tggacgattc aaggcttgct tcataaacca
     3181 aggcaagtaa tagagattgg agtctctaag aaagtagttc ctactgaatc aaaggccatg
     3241 gagtcaaaaa ttcagatcga ggatctaaca gaactcgccg tgaagactgg cgaacagttc
     3301 atacagagtc ttttacgact caatgacaag aagaaaatct tcgtcaacat ggtggagcac
     3361 gacactctcg tctactccaa gaatatcaaa gatacagtct cagaagacca aagggctatt
     3421 gagacttttc aacaaagggt aatatcggga aacctcctcg gattccattg cccagctatc
     3481 tgtcacttca tcaaaaggac agtagaaaag gaaggtggca cctacaaatg ccatcattgc
     3541 gataaaggaa aggctatcgt tcaagatgcc cctgccgaca gtggtcccaa agatggaccc
     3601 ccacccacga ggagcatcgt ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg
     3661 gattgatgtg atatctccac tgacgtaagg gatgacgcac aatcccacta tccttcgcaa
     3721 gacccttcct ctatataagg aagttcattt catttggaga ggactccggt atttttacaa
     3781 caattaccac aacaaaacaa acaacaaaca acattacaat ttactattct agtcgaaatg
     3841 gaacgagcta tacaaggaaa cgacgctagg gaacaagcta acagtgaacg ttgggatgga
     3901 ggatcaggag gtaccacttc tcccttcaaa cttcctgacg aaagtccgag ttggactgag
     3961 tggcggctac ataacgatga gaccaattcg aatcaagata atccccttgg tttcaaggaa
     4021 agctggggtt tcgggaaagt tgtatttaag agatatctca gatacgacag gacggaagct
     4081 tcactgcaca gagtccttgg atcttggacg ggagattcgg ttaactatgc agcatctcga
     4141 tttttcggtt tcgaccagat cggatgtacc tatagtattc ggtttcgagg agttagtatc
     4201 accgtttctg gaggctctcg aactcttcag catctctgtg agatggcaat tcggtctaag
     4261 caagaactgc tacagcttgc cccaatcgaa gtggaaagta atgtatcaag aggatgccct
     4321 gaaggtactg aaaccttcga aaaagaaagc gagtgagctt ggaatggatc ttcgatcccg
     4381 atcgttcaaa catttggcaa taaagtttct taagattgaa tcctgttgcc ggtcttgcga
     4441 cgattatcat ataatttctg ttgaattacg ttaagcatgt aataattaac atgtaatgca
     4501 tgacgttatt tatgagatgg gtttttatga ttagagtccc gcaattatac atttaatacg
     4561 cgatagaaaa caaaatatag cgcgcaaact aggataaatt atcgcgcgcg gtgtcatcta
     4621 tgttactaga tcgggaattg ccaagctaat tcttgaagac gaaagggcct cgtgatacgc
     4681 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     4741 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4801 ccgctcatga gacaataacc ctgataaatg cttcaataat gggaccgact cgcgcttgag
     4861 acgaagcttg acaggatata ttggcgggta aactaagtcg ctgtatgtgt ttgtttgaga
     4921 tctcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4981 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5041 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5101 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tccgtatgtg
     5161 tttgtttgag atctcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5221 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5281 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5341 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5401 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5461 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5521 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5581 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5641 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     5701 ctgaagccag ttaccttcgg aagaagagtt ggtagctctt gatccggcaa acaaaccacc
     5761 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5821 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5881 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5941 aaatgaagtt ttaaatcaat ctaaagtata tatgtgtaac attggtctag tgattagaaa
     6001 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
     6061 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
     6121 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
     6181 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     6241 ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta
     6301 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
     6361 gcgagtcgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
     6421 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     6481 aatacctgga atgctgtttt ccctgggatc gcagtggtga gtaaccatgc atcatcagga
     6541 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
     6601 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     6661 ggcgcatcgg gcttcccata caatcggtag attgtcgcac ctgattgccc gacattatcg
     6721 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggccttgag
     6781 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca
     6841 gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt
     6901 tgagacacaa cgtggctttg ttgaataaat cgaacttttg ctgagttgaa ggatcagatc
     6961 acgcatcttc ccgacaacgc agaccgttcc gtggcaaagc aaaagttcaa aatcaccaac
     7021 tggtccacct acaacaaagc tctcatcaac cgtggctccc tcactttctg gctggatgat
     7081 ggggcgattc aggcgatccc catccaacag cccgccgtcg agcgggcttt tttatccccg
     7141 gaagcctgtg gatagagggt agttatccac gtgaaaccgc taatgccccg caaagccttg
     7201 attcacgggg ctttccggcc cgctccaaaa actatccacg tgaaatcgct aatcagggta
     7261 cgtgaaatcg ctaatcggag tacgtgaaat cgctaataag gtcacgtgaa atcgctaatc
     7321 aaaaaggcac gtgagaacgc taatagccct ttcagatcaa cagcttgcaa acacccctcg
     7381 ctccggcaag tagttacagc aagtagtatg ttcaattagc ttttcaatta tgaatatata
     7441 tatcaattat tggtcgccct tggcttgtgg acaatgcgct acgcgcaccg gctccgcccg
     7501 tggacaaccg caagcggttg cccaccgtcg agcgcctttg cccacaaccc ggcggccggc
     7561 cgcaacagat cgttttataa attttttttt ttgaaaaaga aaaagcccga aaggcggcaa
     7621 cctctcgggc ttctggattt ccgatccccg gaattagatc ttggcaggat atattgtggt
     7681 gtaacaagct tcgtctcagt ca
//
