LOCUS       sequence-209343-        7921 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plant expression as translational fusions to FLUC with RLUC 
            normalization.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7921)
  AUTHORS   Sole-Gil A, Urbez C, Ferrando A, Blazquez MA
  TITLE     Quantification of Xylem-Specific Thermospermine-Dependent 
            Translation of SACL Transcripts with Dual Luciferase Reporter 
            System.
  JOURNAL   Methods Mol Biol. 2024;2722:79-87. doi: 10.1007/978-1-0716-3477-6_6.
  PUBMED    37897601
REFERENCE   2  (bases 1 to 7921)
  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: "Methods Mol
            Biol."; date: "2024"; volume: "2722:79-87. doi"; pages: " 
            10.1007/978-1-0716-3477-6_6"
COMMENT     Sequence Label: pGreen2x35SLUC
FEATURES             Location/Qualifiers
     source          1..7921
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(5..21)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5..21)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             53..1705
                     /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(119..137)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1767..1943
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    1953..1977
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(2068..2656)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2148..2167)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2827..3642)
                     /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(2827..3642)
                     /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     3550..3569
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3634..3653
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     mobile_element  3698..4465
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     rep_origin      4710..5145
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    5272..5294
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     promoter        5356..5701
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             5720..6655
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    6710..6886
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     7035..7057
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7049..7066
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7050..7066
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7076..7095
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7076..7094
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     7119..7135
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     7120..7136
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7567..7911
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
ORIGIN
        1 gggggatcca ctagttctag agcggccgcc accgcggtgg agatcgaatt ccatggaaga
       61 cgccaaaaac ataaagaaag gcccggcgcc attctatccg ctggaagatg gaaccgctgg
      121 agagcaactg cataaggcta tgaagagata cgccctggtt cctggaacaa ttgcttttac
      181 agatgcacat atcgaggtgg acatcactta cgctgagtac ttcgaaatgt ccgttcggtt
      241 ggcagaagct atgaaacgat atgggctgaa tacaaatcac agaatcgtcg tatgcagtga
      301 aaactctctt caattcttta tgccggtgtt gggcgcgtta tttatcggag ttgcagttgc
      361 gcccgcgaac gacatttata atgaacgtga attgctcaac agtatgggca tttcgcagcc
      421 taccgtggtg ttcgtttcca aaaaggggtt gcaaaaaatt ttgaacgtgc aaaaaaagct
      481 cccaatcatc caaaaaatta ttatcatgga ttctaaaacg gattaccagg gatttcagtc
      541 gatgtacacg ttcgtcacat ctcatctacc tcccggtttt aatgaatacg attttgtgcc
      601 agagtccttc gatagggaca agacaattgc actgatcatg aactcctctg gatctactgg
      661 tctgcctaaa ggtgtcgctc tgcctcatag aactgcctgc gtgagattct cgcatgccag
      721 agatcctatt tttggcaatc aaatcattcc ggatactgcg attttaagtg ttgttccatt
      781 ccatcacggt tttggaatgt ttactacact cggatatttg atatgtggat ttcgagtcgt
      841 cttaatgtat agatttgaag aagagctgtt tctgaggagc cttcaggatt acaagattca
      901 aagtgcgctg ctggtgccaa ccctattctc cttcttcgcc aaaagcactc tgattgacaa
      961 atacgattta tctaatttac acgaaattgc ttctggtggc gctcccctct ctaaggaagt
     1021 cggggaagcg gttgccaaga ggttccatct gccaggtatc aggcaaggat atgggctcac
     1081 tgagactaca tcagctattc tgattacacc cgagggggat gataaaccgg gcgcggtcgg
     1141 taaagttgtt ccattttttg aagcgaaggt tgtggatctg gataccggga aaacgctggg
     1201 cgttaatcaa agaggcgaac tgtgtgtgag aggtcctatg attatgtccg gttatgtaaa
     1261 caatccggaa gcgaccaacg ccttgattga caaggatgga tggctacatt ctggagacat
     1321 agcttactgg gacgaagacg aacacttctt catcgttgac cgcctgaagt ctctgattaa
     1381 gtacaaaggc tatcaggtgg ctcccgctga attggaatcc atcttgctcc aacaccccaa
     1441 catcttcgac gcaggtgtcg caggtcttcc cgacgatgac gccggtgaac ttcccgccgc
     1501 cgttgttgtt ttggagcacg gaaagacgat gacggaaaaa gagatcgtgg attacgtcgc
     1561 cagtcaagta acaaccgcga aaaagttgcg cggaggagtt gtgtttgtgg acgaagtacc
     1621 gaaaggtctt accggaaaac tcgacgcaag aaaaatcaga gagatcctca taaaggccaa
     1681 gaagggcgga aagatcgccg tgtaattcta gagaattcgg tacgctgaaa tcaccagtct
     1741 ctctctacaa atctatctct ctctattttc tccataaata atgtgtgagt agtttcccga
     1801 taagggaaat tagggttctt atagggtttc gctcatgtgt tgagcatata agaaaccctt
     1861 agtatgtatt tgtatttgta aaatacttct atcaataaaa tttctaattc ctaaaaccaa
     1921 aatccagtac taaaatccag atccactagc cttgacagga tatattggcg ggtaaactaa
     1981 gtcgctgtat gtgtttgttt gagatctcat gtgagcaaaa ggccagcaaa aggccaggaa
     2041 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2101 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2161 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2221 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2281 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2341 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2401 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2461 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     2521 tatctgcgct ctgctgaagc cagttacctt cggaagaaga gttggtagct cttgatccgg
     2581 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2641 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2701 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     2761 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgtgt aacattggtc
     2821 tagtgattag aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt
     2881 atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca
     2941 gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat
     3001 acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt
     3061 gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac
     3121 aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg
     3181 tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg
     3241 aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc
     3301 aggatattct tctaatacct ggaatgctgt tttccctggg atcgcagtgg tgagtaacca
     3361 tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag
     3421 ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt
     3481 cagaaacaac tctggcgcat cgggcttccc atacaatcgg tagattgtcg cacctgattg
     3541 cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa
     3601 tcgcggcctt gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact
     3661 gtttatgtaa gcagacagtt ttattgttca tgatgatggt gatgctgcca acttactgat
     3721 ttagtgtatg atggtgtttt tgaggtgctc cagtggcttc tgtttctatc agctgtccct
     3781 cctgttcagc tactgacggg gtggtgcgta acggcaaaag caccgccgga catcagcgct
     3841 atctctgctc tcactgccgt aaaacatggc aactgcagtt cacttacacc gcttctcaac
     3901 ccggtacgca ccagaaaatc attgatatgg ccatgaatgg cgttggatgc cgggcaacag
     3961 cccgcattat gggcgttggc ctcaacacga ttttacgtca cttaaaaaac tcaggccgca
     4021 gtcggtaacc tcgcgcatac agccgggcag tgacgtcatc gtctgcgcgg aaatggacga
     4081 acagtggggc tatgtcgggg ctaaatcgcg ccagcgctgg ctgttttacg cgtatgacag
     4141 tctccggaag acggttgttg cgcacgtatt cggtgaacgc actatggcga cgctggggcg
     4201 tcttatgagc ctgctgtcac cctttgacgt ggtgatatgg atgacggatg gctggccgct
     4261 gtatgaatcc cgcctgaagg gaaagctgca cgtaatcagc aagcgatata cgcagcgaat
     4321 tgagcggcat aacctgaatc tgaggcagca cctggcacgg ctgggacgga agtcgctgtc
     4381 gttctcaaaa tcggtggagc tgcatgacaa agtcatcggg cattatctga acataaaaca
     4441 ctatcaataa gttggagtca ttacccatga tgatatattt ttatcttgtg caatgtaaca
     4501 tcagagattt tgagacacaa cgtggctttg ttgaataaat cgaacttttg ctgagttgaa
     4561 ggatcagatc acgcatcttc ccgacaacgc agaccgttcc gtggcaaagc aaaagttcaa
     4621 aatcaccaac tggtccacct acaacaaagc tctcatcaac cgtggctccc tcactttctg
     4681 gctggatgat ggggcgattc aggcgatccc catccaacag cccgccgtcg agcgggcttt
     4741 tttatccccg gaagcctgtg gatagagggt agttatccac gtgaaaccgc taatgccccg
     4801 caaagccttg attcacgggg ctttccggcc cgctccaaaa actatccacg tgaaatcgct
     4861 aatcagggta cgtgaaatcg ctaatcggag tacgtgaaat cgctaataag gtcacgtgaa
     4921 atcgctaatc aaaaaggcac gtgagaacgc taatagccct ttcagatcaa cagcttgcaa
     4981 acacccctcg ctccggcaag tagttacagc aagtagtatg ttcaattagc ttttcaatta
     5041 tgaatatata tatcaattat tggtcgccct tggcttgtgg acaatgcgct acgcgcaccg
     5101 gctccgcccg tggacaaccg caagcggttg cccaccgtcg agcgcctttg cccacaaccc
     5161 ggcggccggc cgcaacagat cgttttataa attttttttt ttgaaaaaga aaaagcccga
     5221 aaggcggcaa cctctcgggc ttctggattt ccgatccccg gaattagatc ttggcaggat
     5281 atattgtggt gtaacgttat cgtaccccta ctccaaaaat gtcaaagata cagtctcaga
     5341 agaccaaagg gctattgaga cttttcaaca aagggtaatt tcgggaaacc tcctcggatt
     5401 ccattgccca gctatctgtc acttcatcga aaggacagta gaaaaggaag gtggctccta
     5461 caaatgccat cattgcgata aaggaaaggc tatcattcaa gatgcctctg ccgacagtgg
     5521 tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagacg ttccaaccac
     5581 gtcttcaaag caagtggatt gatgtgacat ctccactgac gtaagggatg acgcacaatc
     5641 ccactatcct tcgcaagacc cttcctctat ataaggaagt tcatttcatt tggagaggac
     5701 agcccaagct ctagccacca tgacttcgaa agtttatgat ccagaacaaa ggaaacggat
     5761 gataactggt ccgcagtggt gggccagatg taaacaaatg aatgttcttg attcatttat
     5821 taattattat gattcagaaa aacatgcaga aaatgctgtt atttttttac atggtaacgc
     5881 ggcctcttct tatttatggc gacatgttgt gccacatatt gagccagtag cgcggtgtat
     5941 tataccagac cttattggta tgggcaaatc aggcaaatct ggtaatggtt cttataggtt
     6001 acttgatcat tacaaatatc ttactgcatg gtttgaactt cttaatttac caaagaagat
     6061 catttttgtc ggccatgatt ggggtgcttg tttggcattt cattatagct atgagcatca
     6121 agataagatc aaagcaatag ttcacgctga aagtgtagta gatgtgattg aatcatggga
     6181 tgaatggcct gatattgaag aagatattgc gttgatcaaa tctgaagaag gagaaaaaat
     6241 ggttttggag aataacttct tcgtggaaac catgttgcca tcaaaaatca tgagaaagtt
     6301 agaaccagaa gaatttgcag catatcttga accattcaaa gagaaaggtg aagttcgtcg
     6361 tccaacatta tcatggcctc gtgaaatccc gttagtaaaa ggtggtaaac ctgacgttgt
     6421 acaaattgtt aggaattata atgcttatct acgtgcaagt gatgatttac caaaaatgtt
     6481 tattgaatcg gacccaggat tcttttccaa tgctattgtt gaaggtgcca agaagtttcc
     6541 taatactgaa tttgtcaaag taaaaggtct tcatttttcg caagaagatg cacctgatga
     6601 aatgggaaaa tatatcaaat cgttcgttga gcgagttctc aaaaatgaac aataattcta
     6661 gggtacgctg aaatcaccag tctctctcta caaatctatc tctctctatt ttctccataa
     6721 ataatgtgtg agtagtttcc cgataaggga aattagggtt cttatagggt ttcgctcatg
     6781 tgttgagcat ataagaaacc cttagtatgt atttgtattt gtaaaatact tctatcaata
     6841 aaatttctaa ttcctaaaac caaaatccag tactaaaatc cagatcgata acattaacgc
     6901 ttacaatttc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     6961 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     7021 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaata
     7081 cgactcacta tagggcgaat tgggtaccgg gccccccctc gaggtcgacg gtatcgataa
     7141 gcttgatatc gaattcctgc aggtcaacat ggtggagcac gacacacttg tctactccaa
     7201 aaatatcaaa gatacagtct cagaagacca aagggcaatt gagacttttc aacaaagggt
     7261 aatatccgga aacctcctcg gattccattg cccagctatc tgtcacttta ttgtgaagat
     7321 agtggaaaag gaaggtggct cctacaaatg ccatcattgc gataaaggaa aggccatcgt
     7381 tgaagatgcc tctgccgaca gtggtcccaa agatggaccc ccacccacga ggagcatcgt
     7441 ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg gattgatgtg ataacatggt
     7501 ggagcacgac acacttgtct actccaaaaa tatcaaagat acagtctcag aagaccaaag
     7561 ggcaattgag acttttcaac aaagggtaat atccggaaac ctcctcggat tccattgccc
     7621 agctatctgt cactttattg tgaagatagt ggaaaaggaa ggtggctcct acaaatgcca
     7681 tcattgcgat aaaggaaagg ccatcgttga agatgcctct gccgacagtg gtcccaaaga
     7741 tggaccccca cccacgagga gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa
     7801 gcaagtggat tgatgtgata tctccactga cgtaagggat gacgcacaat cccactatcc
     7861 ttcgcaagac ccttcctcta tataaggaag ttcatttcat ttggagagga cctcgagaat
     7921 t
//
