LOCUS       sequence-242089-        7695 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  binary plant vector for transient fLUC (with intron) expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7695)
  AUTHORS   Mortensen S, Bernal-Franco D, Cole LF, Sathitloetsakun S, Cram EJ, 
            Lee-Parsons CWT
  TITLE     EASI Transformation: An Efficient Transient Expression Method for 
            Analyzing Gene Function in Catharanthus roseus Seedlings.
  JOURNAL   Front Plant Sci. 2019 Jun 11;10:755. doi: 10.3389/fpls.2019.00755. 
            eCollection 2019.
  PUBMED    31263474
REFERENCE   2  (bases 1 to 7695)
  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: "Front Plant
            Sci."; date: "2019-06-11"; volume: "10:755. doi"; pages: " 
            10.3389/fpls.2019.00755. eCollection 2019"
COMMENT     Sequence Label: pSB97 - pL2_pVS1_2x35S::fLUC-I::tNOS
FEATURES             Location/Qualifiers
     source          1..7695
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        426..771
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     714..734
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     complement(928..946)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     intron          1006..1194
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     terminator      2716..2968
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    3053..3077
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3357..3986
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4415..5488
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGASSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      5554..5748
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        5810..5914
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5915..6709
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      6880..7468
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7369..7388
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7604..7628
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 gtgccgaatt cggatccgga ggtcaacatg gtggagcacg acactctggt ctactccaaa
       61 aatgtcaaag atacagtctc agaagatcaa agggctattg agacttttca acaaaggata
      121 atttcgggaa acctcctcgg attccattgc ccagctatct gtcacttcat cgaaaggaca
      181 gtagaaaagg aaggtggctc ctacaaatgc catcattgcg ataaaggaaa ggctatcatt
      241 caagatctct ctgccgacag tggtcccaaa gatggacccc cacccacgag gagcatcgtg
      301 gaaaaagaag aggttccaac cacgtctaca aagcaagtgg attgatgtga taacatggtg
      361 gagcacgaca ctctggtcta ctccaaaaat gtcaaagata cagtctcaga agatcaaagg
      421 gctattgaga cttttcaaca aaggataatt tcgggaaacc tcctcggatt ccattgccca
      481 gctatctgtc acttcatcga aaggacagta gaaaaggaag gtggctccta caaatgccat
      541 cattgcgata aaggaaaggc tatcattcaa gatctctctg ccgacagtgg tcccaaagat
      601 ggacccccac ccacgaggag catcgtggaa aaagaagagg ttccaaccac gtctacaaag
      661 caagtggatt gatgtgacat ctccactgac gtaagggatg acgcacaatc ccactatcct
      721 tcgcaagacc cttcctctat ataaggaagt tcatttcatt tggagaggac acgctcgagt
      781 ataagagctc atttttacaa caattaccaa caacaacaaa caacaaacaa cattacaatt
      841 acatttacaa ttatcgatac aatggaggac gccaaaaaca taaagaaagg cccggcgcca
      901 ttctatcctc tagaggatgg aaccgctgga gagcaactgc ataaggctat gaagagatac
      961 gccctggttc ctggaacaat tgcttttaca gatgcacata tcgaggtaag tttctgcttc
     1021 tacctttgat atatatataa taattatcat taattagtag taatataata tttcaaatat
     1081 ttttttcaaa ataaaagaat gtagtatata gcaattgctt ttctgtagtt tataagtgtg
     1141 tatattttaa tttataactt ttctaatata tgaccaaaat ttgttgatgt gcaggtgaac
     1201 atcacgtacg cggaatactt cgaaatgtcc gttcggttgg cagaagctat gaaacgatat
     1261 gggctgaata caaatcacag aatcgtcgta tgcagtgaaa actctcttca attctttatg
     1321 ccggtgttgg gcgcgttatt tatcggagtt gcagttgcgc ccgcgaacga catttataat
     1381 gaacgtgaat tgctcaacag tatgaacatt tcgcagccta ccgtagtgtt tgtttccaaa
     1441 aaggggttgc aaaaaatttt gaacgtgcaa aaaaaattac caataatcca gaaaattatt
     1501 atcatggatt ctaaaacgga ttaccaggga tttcagtcga tgtacacgtt cgtcacatct
     1561 catctacctc ccggttttaa tgaatacgat tttgtaccag agtcctttga tcgtgacaaa
     1621 acaattgcac tgataatgaa ttcctctgga tctactgggt tacctaaggg tgtggccctt
     1681 ccgcatagaa ctgcctgcgt cagattctcg catgccagag atcctatttt tggcaatcaa
     1741 atcattccgg atactgcgat tttaagtgtt gttccattcc atcacggttt tggaatgttt
     1801 actacactcg gatatttgat atgtggattt cgagtcgtct taatgtatag atttgaagaa
     1861 gagctgtttt tacgatccct tcaggattac aaaattcaaa gtgcgttgct agtaccaacc
     1921 ctattttcat tcttcgccaa aagcactctg attgacaaat acgatttatc taatttacac
     1981 gaaattgctt ctgggggcgc acctctttcg aaagaagtcg gggaagcggt tgcaaaacgc
     2041 ttccatcttc cagggatacg acaaggatat gggctcactg agactacatc agctattctg
     2101 attacacccg agggggatga taaaccgggc gcggtcggta aagttgttcc attttttgaa
     2161 gcgaaggttg tggatctgga taccgggaaa acgctgggcg ttaatcagag aggcgaatta
     2221 tgtgtcagag gacctatgat tatgtccggt tatgtaaaca atccggaagc gaccaacgcc
     2281 ttgattgaca aggatggatg gctacattct ggagacatag cttactggga cgaggacgaa
     2341 cacttcttca tagttgaccg cttgaagtct ttaattaaat acaaaggata tcaggtggcc
     2401 cccgctgaat tggaatcgat attgttacaa caccccaaca tcttcgacgc gggcgtggca
     2461 ggtctgcccg acgatgacgc cggtgaactt cccgccgccg ttgttgtttt ggagcacgga
     2521 aagacgatga cggaaaaaga gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa
     2581 aagttgcgcg gaggagttgt gtttgtggac gaagtaccga aaggtcttac cggaaaactc
     2641 gacgcaagaa aaatcagaga gatcctcata aaggccaaga agggcggaaa gtccaaattg
     2701 taagcttgtc aagcagatcg ttcaaacatt tggcaataaa gtttcttaag attgaatcct
     2761 gttgccggtc ttgcgatgat tatcatataa tttctgttga attacgttaa gcatgtaata
     2821 attaacatgt aatgcatgac gttatttatg agatgggttt ttatgattag agtcccgcaa
     2881 ttatacattt aatacgcgat agaaaacaaa atatagcgcg caaactagga taaattatcg
     2941 cgcgcggtgt catctatgtt actagatcga cgctgcaagg ttttgacggg tttactagag
     3001 gatgcacatg tgaccgaggg acacgaagtg atccgtttaa actatcagtg tttgacagga
     3061 tatattggcg ggtaaaccta agagaaaaga gcgtttatta gaataatcgg atatttaaaa
     3121 gggcgtgaaa aggtttatcc gttcgtccat ttgtatgtgc cagccgtgcg gctgcatgaa
     3181 atcctggccg gtttgtctga tgccaagctg gcggcctggc cggccagctt ggccgctgaa
     3241 gaaaccgagc gccgccgtct aaaaaggtga tgtgtatttg agtaaaacag cttgcgtcat
     3301 gcggtcgctg cgtatatgat gcgatgagta aataaacaaa tacgcaaggg gaacgcatga
     3361 aggttatcgc tgtacttaac cagaaaggcg ggtcaggcaa gacgaccatc gcaacccatc
     3421 tagcccgcgc cctgcaactc gccggggccg atgttctgtt agtcgattcc gatccccagg
     3481 gcagtgcccg cgattgggcg gccgtgcggg aagatcaacc gctaaccgtt gtcggcatcg
     3541 accgcccgac gattgaccgc gacgtgaagg ccatcggccg gcgcgacttc gtagtgatcg
     3601 acggagcgcc ccaggcggcg gacttggctg tgtccgcgat caaggcagcc gacttcgtgc
     3661 tgattccggt gcagccaagc ccttacgaca tatgggccac cgccgacctg gtggagctgg
     3721 ttaagcagcg cattgaggtc acggatggaa ggctacaagc ggcctttgtc gtgtcgcggg
     3781 cgatcaaagg cacgcgcatc ggcggtgagg ttgccgaggc gctggccggg tacgagctgc
     3841 ccattcttga gtcccgtatc acgcagcgcg tgagctaccc aggcactgcc gccgccggca
     3901 caaccgttct tgaatcagaa cccgagggcg acgctgcccg cgaggtccag gcgctggccg
     3961 ctgaaattaa atcaaaactc atttgagtta atgaggtaaa gagaaaatga gcaaaagcac
     4021 aaacacgcta agtgccggcc gtccgagcgc acgcagcagc aaggctgcaa cgttggccag
     4081 cctggcagac acgccagcca tgaagcgggt caactttcag ttgccggcgg aggatcacac
     4141 caagctgaag atgtacgcgg tacgccaagg caagaccatt accgagctgc tatctgaata
     4201 catcgcgcag ctaccagagt aaatgagcaa atgaataaat gagtagatga attttagcgg
     4261 ctaaaggagg cggcatggaa aatcaagaac aaccaggcac cgacgccgtg gaatgcccca
     4321 tgtgtggagg aacgggcggt tggccaggcg taagcggctg ggttgtctgc cggccctgca
     4381 atggcactgg aacccccaag cccgaggaat cggcgtgagc ggtcgcaaac catccggccc
     4441 ggtacaaatc ggcgcggcgc tgggtgatga cctggtggag aagttgaagg ccgcgcaggc
     4501 cgcccagcgg caacgcatcg aggcagaagc acgccccggt gaatcgtggc aagcggccgc
     4561 tgatcgaatc cgcaaagaat cccggcaacc gccggcagcc ggtgcgtcgt cgattaggaa
     4621 gccgcccaag ggcgacgagc aaccagattt tttcgttccg atgctctatg acgtgggcac
     4681 ccgcgatagt cgcagcatca tggacgtggc cgttttccgt ctgtcgaagc gtgaccgacg
     4741 agctggcgag gtgatccgct acgagcttcc agacgggcac gtagaggttt ccgcagggcc
     4801 ggccggcatg gccagtgtgt gggattacga cctggtactg atggcggttt cccatctaac
     4861 cgaatccatg aaccgatacc gggaagggaa gggagacaag cccggccgcg tgttccgtcc
     4921 acacgttgcg gacgtactca agttctgccg gcgagccgat ggcggaaagc agaaagacga
     4981 cctggtagaa acctgcattc ggttaaacac cacgcacgtt gccatgcagc gtacgaagaa
     5041 ggccaagaac ggccgcctgg tgacggtatc cgagggtgaa gccttgatta gccgctacaa
     5101 gatcgtaaag agcgaaaccg ggcggccgga gtacatcgag atcgagctag ctgattggat
     5161 gtaccgcgag atcacagaag gcaagaaccc ggacgtgctg acggttcacc ccgattactt
     5221 tttgatcgat cccggcatcg gccgttttct ctaccgcctg gcacgccgcg ccgcaggcaa
     5281 ggcagaagcc agatggttgt tcaagacgat ctacgaacgc agtggcagcg ccggagagtt
     5341 caagaagttc tgtttcaccg tgcgcaagct gatcgggtca aatgacctgc cggagtacga
     5401 tttgaaggag gaggcggggc aggctggccc gatcctagtc atgcgctacc gcaacctgat
     5461 cgagggcgaa gcatccgccg gttcctaatg tacggagcag atgctagggc aaattgccct
     5521 agcaggggaa aaaggtcgaa aaagcttctt tcctgtggat agcacgtaca ttgggaaccc
     5581 aaagccgtac attgggaacc ggaacccgta cattgggaac ccaaagccgt acattgggaa
     5641 ccggtcacac atgtaagtga ctgatataaa agagaaaaaa ggcgattttt ccgcctaaaa
     5701 ctctttaaaa cttattaaaa ctcttaaaac ccgcctggcc tgtgcataac tgtctggcca
     5761 gcgcacagcc gaacagctgc aaaaagcgcc cacatcaagg ctccgagtgc gcggaacccc
     5821 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     5881 ataaatgctt caataatatt gaaaaaggaa gagtatggct aaaatgagaa tatcaccgga
     5941 attgaaaaaa ctgatcgaaa aataccgctg cgtaaaagat acggaaggaa tgtctcctgc
     6001 taaggtatat aagctggtgg gagaaaatga aaacctatat ttaaaaatga cggacagccg
     6061 gtataaaggg accacctatg atgtggaacg ggaaaaggac atgatgctat ggctggaagg
     6121 aaagctgcct gttccaaagg tcctgcactt tgaacggcat gatggctgga gcaatctgct
     6181 catgagtgag gccgatggcg tcctttgctc ggaagagtat gaagatgaac aaagccctga
     6241 aaagattatc gagctgtatg cggagtgcat caggctcttt cactccatcg acatatcgga
     6301 ttgtccctat acgaatagct tagacagccg cttagccgaa ttggattact tactgaataa
     6361 cgatctggcc gatgtggatt gcgaaaactg ggaagaggac actccattta aagatccgcg
     6421 cgagctgtat gattttttaa agacggaaaa gcccgaagag gaacttgtct tttcccacgg
     6481 cgacctggga gacagcaaca tctttgtgaa agatggcaaa gtaagtggct ttattgatct
     6541 tgggagaagc ggcagggcgg acaagtggta tgacattgcc ttctgcgtcc ggtcgctcag
     6601 ggaggatatc ggggaagaac agtatgtcga gctatttttt gacttactgg ggatcaagcc
     6661 tgattgggag aaaataaaat attatatttt actggatgaa ttgttttagc tgtcagacca
     6721 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     6781 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     6841 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6901 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6961 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     7021 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     7081 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     7141 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     7201 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     7261 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     7321 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     7381 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     7441 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     7501 gctcggatct gttggaccgg acagtagtca tggttgatgg gctgcctgta tcgagtggtg
     7561 attttgtgcc gagctgccgg tcggggagct gttggctggc tggtggcagg atatattgtg
     7621 gtgtaaacaa attgacgctt agacaactta ataacacatt gcggacgttt ttaatgtact
     7681 ggggttgaac actct
//
