LOCUS       sequence-242250-        7597 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  binary plant vector for transient expression of a Renilla luciferase
            (rLUC, with intron) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7597)
  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 7597)
  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: pSB166 - pL2_pSB90_tMAS::rLUC-I::pMAS
FEATURES             Location/Qualifiers
     source          1..7597
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    163..187
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     promoter        253..633
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     terminator      1676..1928
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     misc_feature    2055..2079
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(2215..2803)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2295..2314)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4128..4922)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(4923..5027)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5089..5283
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(5349..6422)
                     /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"
     CDS             complement(6851..7480)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
        1 tcagcggcca agctggccgg ccaggccgcc agcttggcat cagacaaacc ggccaggatt
       61 tcatgcagcc gcacggctgg cacatacaaa tggacgaacg gataaacctt ttcacgccct
      121 tttaaatatc cgattattct aataaacgct cttttctctt aggtttaccc gccaatatat
      181 cctgtcaaac actgatagtt taaacggatc acttcgtgtc cctcggtcac atgtgcatcc
      241 tcgtaaggag atttttcaaa tcagtgcgca agacgtgacg taagtatccg agtcagtttt
      301 tatttttcta ctaatttggt cgtttatttc ggcgtgtagg acatggcaac cgggcctgaa
      361 tttcgcgggt attctgtttc tattccaact ttttcttgat ccgcagccat taacgacttt
      421 tgaatagata cgctgacacg ccaagcctcg ctagtcaaaa gtgtaccaaa caacgcttta
      481 cagcaagaac ggaatgcgcg tgacgctcgc ggtgacgcca tttcgccttt tcagaaatgg
      541 ataaatagcc ttgcttccta ttatatcttc ccaaattacc aatacattac actagcatct
      601 gaatttcata accaatctcg atacaccaaa tcgaatgact tctaaggttt atgatcctga
      661 acaaagaaag agaatgatta ctggacctca atggtgggct agatgtaagc aaatgaatgt
      721 tcttgattct tttattaatt attatgattc tgaaaagcat gctgaaaatg ctgttatttt
      781 tcttcatgga aatgctgctt cttcttatct ttggagacat gttgttccac atattgaacc
      841 agttgctaga tgtattattc cagatcttat tggtatggga aagtctggta agtctggtaa
      901 tggttcttat agacttcttg atcattataa gtatcttact gcttggtttg aacttcttaa
      961 tcttccaaag aagattattt ttgttggaca tgattggggt gcttgtttgg cttttcatta
     1021 ttcttatgaa catcaagata agattaaggc tattgttcat gctgaatctg ttgttgatgt
     1081 tattgaatct tgggatgaat ggcctgatat tgaagaagat attgctttga ttaagtctga
     1141 agaaggagaa aagatggttt tggaaaataa tttctttgtt gaaactatgt tgccatctaa
     1201 gattatgaga aagcttgaac cagaagaatt tgctgcttat cttgaaccat ttaaggaaaa
     1261 gggtgaagtt agaaggccaa ctctttcttg gcctagagaa attcctcttg ttaagggtgg
     1321 taagcctgat gttgttcaaa ttgttagaaa ttataatgct tatcttagag cttctgatga
     1381 tcttccaaag atgtttattg aatctgatcc aggattcttt tctaatgcta ttgttgaagg
     1441 tgctaagaag tttcctaata ctgaatttgt taaggtactt catgcttcaa cgtgtaactt
     1501 aagagatact gtgtgaaatt ttatatttcc atacatttgc ttgacctttg ctttttgtca
     1561 atttttttcc ccttacaggt taagggactt catttttctc aagaagatgc tcctgatgaa
     1621 atgggaaagt atattaagtc ttttgttgaa agagttctta agaatgaaca atgagcttgg
     1681 actcccatgt tggcaaaggc aaccaaacaa acaatgaatg atccgctcct gcatatgggg
     1741 cggtttgagt atttcaactg ccatttgggc tgaattgtag acatgctcct gtcagaaatt
     1801 ccgtgatctt actcaatatt cagtaatctc ggccaatatc ctaaatgtgc gtggctttat
     1861 ctgtctttgt attgtttcat caattcatgt aacgtttgct tttcttatga attttcaaat
     1921 aaattatccg ctgagctcga attctagtaa acccgtcaaa accttgcgag aggtgataca
     1981 tcggcacaga gtgttcaacc ccagtacatt aaaaacgtcc gcaatgtgtt attaagttgt
     2041 ctaagcgtca atttgtttac accacaatat atcctgccac cagccagcca acagctcccc
     2101 gaccggcagc tcggcacaaa atcaccactc gatacaggca gcccatcaac catgactact
     2161 gtccggtcca acagatccga gcaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2221 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2281 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2341 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2401 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2461 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2521 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2581 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2641 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2701 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2761 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2821 ttctacgggg tctgacgctc agtgagatca cgagccagta tcctgacgaa tgggttctcg
     2881 tccaacgccc ctttatcgac agtatgtgaa tttgcagtgt ggccgcgatc aacgcacact
     2941 ggccttgggt ctgaagggtg atcttcaaaa cgaacccccg tcttggtggt cagtgtgatg
     3001 ggcacatcgc ttactcctca gtcacccttt gagccagata attaagaggc aaatgcaatt
     3061 ggctcaggct gccatcgtcc ccccgtgcga aacctgcacg tccgcgtcaa agaaataacc
     3121 ggcacctctt gctgttttta tcagttgagg gcttgacgga tccgcctcaa gtttgcggcg
     3181 cagcctcaaa atgagaacat ctatactcct gtcataaacc tcctcgtcgc gtactcgact
     3241 ggcaatgaga agttgctcgc gcgatagaac gtcgcggggt ttctctaaaa acgcgaggag
     3301 aagattgaac tcacctgccg taagtttcac ctcaccgcca gcttcggaca tcaagcgacg
     3361 ttgcctgaga ttaagtgtcc agtcagtaaa acaaaaagac cgtcggtctt tggagcggac
     3421 aacgttgggg cgcacgcgca aggcaacccg aatgcgtgct agaaactctc tgatactgaa
     3481 cggcttagcg ataaaatcac ttgctcctag ctcgagtgca acaactttat ccgtctcctc
     3541 aaggcggtcg ccactgataa ttatgattgg aatatcagac tttgccgcca gattacgaac
     3601 gatctcgagc ccatcttcac gacctaaatc tagatcaaca accacgacat cgaccgtcgc
     3661 ggaagagagt actcttgtga actgggtgct gtcggctacc gcggtcactt tgaaggcgtg
     3721 gatcgtaagg tattcaataa taagatgccg catagcgacg tcgtcatcga caagaagaac
     3781 gtgtttcaac ggctcacctt tcaatctaaa atctgaaccc ttgttcacag cgcttgagaa
     3841 attttcacgt gaaggatgta caatcatctc cagctaaatg ggcagttcgt cagaattgcg
     3901 gctgaccgcg gatgacaaaa atgcgaacca agtatttcaa ttttatgaca aaaattctca
     3961 atcgttgtta caagtgaaac gcttcgaggt tacagctaga acgaaaactc acgttaaggg
     4021 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4081 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagcta aaacaattca
     4141 tccagtaaaa tataatattt tattttctcc caatcaggct tgatccccag taagtcaaaa
     4201 aatagctcga catactgttc ttccccgata tcctccctga gcgaccggac gcagaaggca
     4261 atgtcatacc acttgtccgc cctgccgctt ctcccaagat caataaagcc acttactttg
     4321 ccatctttca caaagatgtt gctgtctccc aggtcgccgt gggaaaagac aagttcctct
     4381 tcgggctttt ccgtctttaa aaaatcatac agctcgcgcg gatctttaaa tggagtgtcc
     4441 tcttcccagt tttcgcaatc cacatcggcc agatcgttat tcagtaagta atccaattcg
     4501 gctaagcggc tgtctaagct attcgtatag ggacaatccg atatgtcgat ggagtgaaag
     4561 agcctgatgc actccgcata cagctcgata atcttttcag ggctttgttc atcttcatac
     4621 tcttccgagc aaaggacgcc atcggcctca ctcatgagca gattgctcca gccatcatgc
     4681 cgttcaaagt gcaggacctt tggaacaggc agctttcctt ccagccatag catcatgtcc
     4741 ttttcccgtt ccacatcata ggtggtccct ttataccggc tgtccgtcat ttttaaatat
     4801 aggttttcat tttctcccac cagcttatat accttagcag gagacattcc ttccgtatct
     4861 tttacgcagc ggtatttttc gatcagtttt ttcaattccg gtgatattct cattttagcc
     4921 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     4981 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac tcggagcctt
     5041 gatgtgggcg ctttttgcag ctgttcggct gtgcgctggc cagacagtta tgcacaggcc
     5101 aggcgggttt taagagtttt aataagtttt aaagagtttt aggcggaaaa atcgcctttt
     5161 ttctctttta tatcagtcac ttacatgtgt gaccggttcc caatgtacgg ctttgggttc
     5221 ccaatgtacg ggttccggtt cccaatgtac ggctttgggt tcccaatgta cgtgctatcc
     5281 acaggaaaga agctttttcg acctttttcc cctgctaggg caatttgccc tagcatctgc
     5341 tccgtacatt aggaaccggc ggatgcttcg ccctcgatca ggttgcggta gcgcatgact
     5401 aggatcgggc cagcctgccc cgcctcctcc ttcaaatcgt actccggcag gtcatttgac
     5461 ccgatcagct tgcgcacggt gaaacagaac ttcttgaact ctccggcgct gccactgcgt
     5521 tcgtagatcg tcttgaacaa ccatctggct tctgccttgc ctgcggcgcg gcgtgccagg
     5581 cggtagagaa aacggccgat gccgggatcg atcaaaaagt aatcggggtg aaccgtcagc
     5641 acgtccgggt tcttgccttc tgtgatctcg cggtacatcc aatcagctag ctcgatctcg
     5701 atgtactccg gccgcccggt ttcgctcttt acgatcttgt agcggctaat caaggcttca
     5761 ccctcggata ccgtcaccag gcggccgttc ttggccttct tcgtacgctg catggcaacg
     5821 tgcgtggtgt ttaaccgaat gcaggtttct accaggtcgt ctttctgctt tccgccatcg
     5881 gctcgccggc agaacttgag tacgtccgca acgtgtggac ggaacacgcg gccgggcttg
     5941 tctcccttcc cttcccggta tcggttcatg gattcggtta gatgggaaac cgccatcagt
     6001 accaggtcgt aatcccacac actggccatg ccggccggcc ctgcggaaac ctctacgtgc
     6061 ccgtctggaa gctcgtagcg gatcacctcg ccagctcgtc ggtcacgctt cgacagacgg
     6121 aaaacggcca cgtccatgat gctgcgacta tcgcgggtgc ccacgtcata gagcatcgga
     6181 acgaaaaaat ctggttgctc gtcgcccttg ggcggcttcc taatcgacga cgcaccggct
     6241 gccggcggtt gccgggattc tttgcggatt cgatcagcgg ccgcttgcca cgattcaccg
     6301 gggcgtgctt ctgcctcgat gcgttgccgc tgggcggcct gcgcggcctt caacttctcc
     6361 accaggtcat cacccagcgc cgcgccgatt tgtaccgggc cggatggttt gcgaccgctc
     6421 acgccgattc ctcgggcttg ggggttccag tgccattgca gggccggcag acaacccagc
     6481 cgcttacgcc tggccaaccg cccgttcctc cacacatggg gcattccacg gcgtcggtgc
     6541 ctggttgttc ttgattttcc atgccgcctc ctttagccgc taaaattcat ctactcattt
     6601 attcatttgc tcatttactc tggtagctgc gcgatgtatt cagatagcag ctcggtaatg
     6661 gtcttgcctt ggcgtaccgc gtacatcttc agcttggtgt gatcctccgc cggcaactga
     6721 aagttgaccc gcttcatggc tggcgtgtct gccaggctgg ccaacgttgc agccttgctg
     6781 ctgcgtgcgc tcggacggcc ggcacttagc gtgtttgtgc ttttgctcat tttctcttta
     6841 cctcattaac tcaaatgagt tttgatttaa tttcagcggc cagcgcctgg acctcgcggg
     6901 cagcgtcgcc ctcgggttct gattcaagaa cggttgtgcc ggcggcggca gtgcctgggt
     6961 agctcacgcg ctgcgtgata cgggactcaa gaatgggcag ctcgtacccg gccagcgcct
     7021 cggcaacctc accgccgatg cgcgtgcctt tgatcgcccg cgacacgaca aaggccgctt
     7081 gtagccttcc atccgtgacc tcaatgcgct gcttaaccag ctccaccagg tcggcggtgg
     7141 cccatatgtc gtaagggctt ggctgcaccg gaatcagcac gaagtcggct gccttgatcg
     7201 cggacacagc caagtccgcc gcctggggcg ctccgtcgat cactacgaag tcgcgccggc
     7261 cgatggcctt cacgtcgcgg tcaatcgtcg ggcggtcgat gccgacaacg gttagcggtt
     7321 gatcttcccg cacggccgcc caatcgcggg cactgccctg gggatcggaa tcgactaaca
     7381 gaacatcggc cccggcgagt tgcagggcgc gggctagatg ggttgcgatg gtcgtcttgc
     7441 ctgacccgcc tttctggtta agtacagcga taaccttcat gcgttcccct tgcgtatttg
     7501 tttatttact catcgcatca tatacgcagc gaccgcatga cgcaagctgt tttactcaaa
     7561 tacacatcac ctttttagac ggcggcgctc ggtttct
//
