LOCUS       sequence-242082-        8849 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 8849)
  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 8849)
  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: pSB96 - pL2_pSB90_2x35S::fLUC-I::tNOS
FEATURES             Location/Qualifiers
     source          1..8849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      182..770
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     671..690
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    906..930
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        1423..1768
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     1711..1731
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     complement(1925..1943)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     intron          2003..2191
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     terminator      3713..3965
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    4050..4074
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             4354..4983
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             5412..6485
                     /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      6551..6745
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        6807..6911
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6912..7706
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
        1 aagacggggg ttcgttttga agatcaccct tcagacccaa ggccagtgtg cgttgatcgc
       61 ggccacactg caaattcaca tactgtcgat aaaggggcgt tggacgagaa cccattcgtc
      121 aggatactgg ctcgtgatct cactgagcgt cagaccccgt agaaaagatc aaaggatctt
      181 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
      241 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
      301 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
      361 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
      421 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
      481 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
      541 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
      601 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
      661 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
      721 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
      781 acgcggcctt tttacggttc ctgctcggat ctgttggacc ggacagtagt catggttgat
      841 gggctgcctg tatcgagtgg tgattttgtg ccgagctgcc ggtcggggag ctgttggctg
      901 gctggtggca ggatatattg tggtgtaaac aaattgacgc ttagacaact taataacaca
      961 ttgcggacgt ttttaatgta ctggggttga acactctgtg ccgaattcgg atccggaggt
     1021 caacatggtg gagcacgaca ctctggtcta ctccaaaaat gtcaaagata cagtctcaga
     1081 agatcaaagg gctattgaga cttttcaaca aaggataatt tcgggaaacc tcctcggatt
     1141 ccattgccca gctatctgtc acttcatcga aaggacagta gaaaaggaag gtggctccta
     1201 caaatgccat cattgcgata aaggaaaggc tatcattcaa gatctctctg ccgacagtgg
     1261 tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagagg ttccaaccac
     1321 gtctacaaag caagtggatt gatgtgataa catggtggag cacgacactc tggtctactc
     1381 caaaaatgtc aaagatacag tctcagaaga tcaaagggct attgagactt ttcaacaaag
     1441 gataatttcg ggaaacctcc tcggattcca ttgcccagct atctgtcact tcatcgaaag
     1501 gacagtagaa aaggaaggtg gctcctacaa atgccatcat tgcgataaag gaaaggctat
     1561 cattcaagat ctctctgccg acagtggtcc caaagatgga cccccaccca cgaggagcat
     1621 cgtggaaaaa gaagaggttc caaccacgtc tacaaagcaa gtggattgat gtgacatctc
     1681 cactgacgta agggatgacg cacaatccca ctatccttcg caagaccctt cctctatata
     1741 aggaagttca tttcatttgg agaggacacg ctcgagtata agagctcatt tttacaacaa
     1801 ttaccaacaa caacaaacaa caaacaacat tacaattaca tttacaatta tcgatacaat
     1861 ggaggacgcc aaaaacataa agaaaggccc ggcgccattc tatcctctag aggatggaac
     1921 cgctggagag caactgcata aggctatgaa gagatacgcc ctggttcctg gaacaattgc
     1981 ttttacagat gcacatatcg aggtaagttt ctgcttctac ctttgatata tatataataa
     2041 ttatcattaa ttagtagtaa tataatattt caaatatttt tttcaaaata aaagaatgta
     2101 gtatatagca attgcttttc tgtagtttat aagtgtgtat attttaattt ataacttttc
     2161 taatatatga ccaaaatttg ttgatgtgca ggtgaacatc acgtacgcgg aatacttcga
     2221 aatgtccgtt cggttggcag aagctatgaa acgatatggg ctgaatacaa atcacagaat
     2281 cgtcgtatgc agtgaaaact ctcttcaatt ctttatgccg gtgttgggcg cgttatttat
     2341 cggagttgca gttgcgcccg cgaacgacat ttataatgaa cgtgaattgc tcaacagtat
     2401 gaacatttcg cagcctaccg tagtgtttgt ttccaaaaag gggttgcaaa aaattttgaa
     2461 cgtgcaaaaa aaattaccaa taatccagaa aattattatc atggattcta aaacggatta
     2521 ccagggattt cagtcgatgt acacgttcgt cacatctcat ctacctcccg gttttaatga
     2581 atacgatttt gtaccagagt cctttgatcg tgacaaaaca attgcactga taatgaattc
     2641 ctctggatct actgggttac ctaagggtgt ggcccttccg catagaactg cctgcgtcag
     2701 attctcgcat gccagagatc ctatttttgg caatcaaatc attccggata ctgcgatttt
     2761 aagtgttgtt ccattccatc acggttttgg aatgtttact acactcggat atttgatatg
     2821 tggatttcga gtcgtcttaa tgtatagatt tgaagaagag ctgtttttac gatcccttca
     2881 ggattacaaa attcaaagtg cgttgctagt accaacccta ttttcattct tcgccaaaag
     2941 cactctgatt gacaaatacg atttatctaa tttacacgaa attgcttctg ggggcgcacc
     3001 tctttcgaaa gaagtcgggg aagcggttgc aaaacgcttc catcttccag ggatacgaca
     3061 aggatatggg ctcactgaga ctacatcagc tattctgatt acacccgagg gggatgataa
     3121 accgggcgcg gtcggtaaag ttgttccatt ttttgaagcg aaggttgtgg atctggatac
     3181 cgggaaaacg ctgggcgtta atcagagagg cgaattatgt gtcagaggac ctatgattat
     3241 gtccggttat gtaaacaatc cggaagcgac caacgccttg attgacaagg atggatggct
     3301 acattctgga gacatagctt actgggacga ggacgaacac ttcttcatag ttgaccgctt
     3361 gaagtcttta attaaataca aaggatatca ggtggccccc gctgaattgg aatcgatatt
     3421 gttacaacac cccaacatct tcgacgcggg cgtggcaggt ctgcccgacg atgacgccgg
     3481 tgaacttccc gccgccgttg ttgttttgga gcacggaaag acgatgacgg aaaaagagat
     3541 cgtggattac gtcgccagtc aagtaacaac cgcgaaaaag ttgcgcggag gagttgtgtt
     3601 tgtggacgaa gtaccgaaag gtcttaccgg aaaactcgac gcaagaaaaa tcagagagat
     3661 cctcataaag gccaagaagg gcggaaagtc caaattgtaa gcttgtcaag cagatcgttc
     3721 aaacatttgg caataaagtt tcttaagatt gaatcctgtt gccggtcttg cgatgattat
     3781 catataattt ctgttgaatt acgttaagca tgtaataatt aacatgtaat gcatgacgtt
     3841 atttatgaga tgggttttta tgattagagt cccgcaatta tacatttaat acgcgataga
     3901 aaacaaaata tagcgcgcaa actaggataa attatcgcgc gcggtgtcat ctatgttact
     3961 agatcgacgc tgcaaggttt tgacgggttt actagaggat gcacatgtga ccgagggaca
     4021 cgaagtgatc cgtttaaact atcagtgttt gacaggatat attggcgggt aaacctaaga
     4081 gaaaagagcg tttattagaa taatcggata tttaaaaggg cgtgaaaagg tttatccgtt
     4141 cgtccatttg tatgtgccag ccgtgcggct gcatgaaatc ctggccggtt tgtctgatgc
     4201 caagctggcg gcctggccgg ccagcttggc cgctgaagaa accgagcgcc gccgtctaaa
     4261 aaggtgatgt gtatttgagt aaaacagctt gcgtcatgcg gtcgctgcgt atatgatgcg
     4321 atgagtaaat aaacaaatac gcaaggggaa cgcatgaagg ttatcgctgt acttaaccag
     4381 aaaggcgggt caggcaagac gaccatcgca acccatctag cccgcgccct gcaactcgcc
     4441 ggggccgatg ttctgttagt cgattccgat ccccagggca gtgcccgcga ttgggcggcc
     4501 gtgcgggaag atcaaccgct aaccgttgtc ggcatcgacc gcccgacgat tgaccgcgac
     4561 gtgaaggcca tcggccggcg cgacttcgta gtgatcgacg gagcgcccca ggcggcggac
     4621 ttggctgtgt ccgcgatcaa ggcagccgac ttcgtgctga ttccggtgca gccaagccct
     4681 tacgacatat gggccaccgc cgacctggtg gagctggtta agcagcgcat tgaggtcacg
     4741 gatggaaggc tacaagcggc ctttgtcgtg tcgcgggcga tcaaaggcac gcgcatcggc
     4801 ggtgaggttg ccgaggcgct ggccgggtac gagctgccca ttcttgagtc ccgtatcacg
     4861 cagcgcgtga gctacccagg cactgccgcc gccggcacaa ccgttcttga atcagaaccc
     4921 gagggcgacg ctgcccgcga ggtccaggcg ctggccgctg aaattaaatc aaaactcatt
     4981 tgagttaatg aggtaaagag aaaatgagca aaagcacaaa cacgctaagt gccggccgtc
     5041 cgagcgcacg cagcagcaag gctgcaacgt tggccagcct ggcagacacg ccagccatga
     5101 agcgggtcaa ctttcagttg ccggcggagg atcacaccaa gctgaagatg tacgcggtac
     5161 gccaaggcaa gaccattacc gagctgctat ctgaatacat cgcgcagcta ccagagtaaa
     5221 tgagcaaatg aataaatgag tagatgaatt ttagcggcta aaggaggcgg catggaaaat
     5281 caagaacaac caggcaccga cgccgtggaa tgccccatgt gtggaggaac gggcggttgg
     5341 ccaggcgtaa gcggctgggt tgtctgccgg ccctgcaatg gcactggaac ccccaagccc
     5401 gaggaatcgg cgtgagcggt cgcaaaccat ccggcccggt acaaatcggc gcggcgctgg
     5461 gtgatgacct ggtggagaag ttgaaggccg cgcaggccgc ccagcggcaa cgcatcgagg
     5521 cagaagcacg ccccggtgaa tcgtggcaag cggccgctga tcgaatccgc aaagaatccc
     5581 ggcaaccgcc ggcagccggt gcgtcgtcga ttaggaagcc gcccaagggc gacgagcaac
     5641 cagatttttt cgttccgatg ctctatgacg tgggcacccg cgatagtcgc agcatcatgg
     5701 acgtggccgt tttccgtctg tcgaagcgtg accgacgagc tggcgaggtg atccgctacg
     5761 agcttccaga cgggcacgta gaggtttccg cagggccggc cggcatggcc agtgtgtggg
     5821 attacgacct ggtactgatg gcggtttccc atctaaccga atccatgaac cgataccggg
     5881 aagggaaggg agacaagccc ggccgcgtgt tccgtccaca cgttgcggac gtactcaagt
     5941 tctgccggcg agccgatggc ggaaagcaga aagacgacct ggtagaaacc tgcattcggt
     6001 taaacaccac gcacgttgcc atgcagcgta cgaagaaggc caagaacggc cgcctggtga
     6061 cggtatccga gggtgaagcc ttgattagcc gctacaagat cgtaaagagc gaaaccgggc
     6121 ggccggagta catcgagatc gagctagctg attggatgta ccgcgagatc acagaaggca
     6181 agaacccgga cgtgctgacg gttcaccccg attacttttt gatcgatccc ggcatcggcc
     6241 gttttctcta ccgcctggca cgccgcgccg caggcaaggc agaagccaga tggttgttca
     6301 agacgatcta cgaacgcagt ggcagcgccg gagagttcaa gaagttctgt ttcaccgtgc
     6361 gcaagctgat cgggtcaaat gacctgccgg agtacgattt gaaggaggag gcggggcagg
     6421 ctggcccgat cctagtcatg cgctaccgca acctgatcga gggcgaagca tccgccggtt
     6481 cctaatgtac ggagcagatg ctagggcaaa ttgccctagc aggggaaaaa ggtcgaaaaa
     6541 gcttctttcc tgtggatagc acgtacattg ggaacccaaa gccgtacatt gggaaccgga
     6601 acccgtacat tgggaaccca aagccgtaca ttgggaaccg gtcacacatg taagtgactg
     6661 atataaaaga gaaaaaaggc gatttttccg cctaaaactc tttaaaactt attaaaactc
     6721 ttaaaacccg cctggcctgt gcataactgt ctggccagcg cacagccgaa cagctgcaaa
     6781 aagcgcccac atcaaggctc cgagtgcgcg gaacccctat ttgtttattt ttctaaatac
     6841 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     6901 aaaggaagag tatggctaaa atgagaatat caccggaatt gaaaaaactg atcgaaaaat
     6961 accgctgcgt aaaagatacg gaaggaatgt ctcctgctaa ggtatataag ctggtgggag
     7021 aaaatgaaaa cctatattta aaaatgacgg acagccggta taaagggacc acctatgatg
     7081 tggaacggga aaaggacatg atgctatggc tggaaggaaa gctgcctgtt ccaaaggtcc
     7141 tgcactttga acggcatgat ggctggagca atctgctcat gagtgaggcc gatggcgtcc
     7201 tttgctcgga agagtatgaa gatgaacaaa gccctgaaaa gattatcgag ctgtatgcgg
     7261 agtgcatcag gctctttcac tccatcgaca tatcggattg tccctatacg aatagcttag
     7321 acagccgctt agccgaattg gattacttac tgaataacga tctggccgat gtggattgcg
     7381 aaaactggga agaggacact ccatttaaag atccgcgcga gctgtatgat tttttaaaga
     7441 cggaaaagcc cgaagaggaa cttgtctttt cccacggcga cctgggagac agcaacatct
     7501 ttgtgaaaga tggcaaagta agtggcttta ttgatcttgg gagaagcggc agggcggaca
     7561 agtggtatga cattgccttc tgcgtccggt cgctcaggga ggatatcggg gaagaacagt
     7621 atgtcgagct attttttgac ttactgggga tcaagcctga ttgggagaaa ataaaatatt
     7681 atattttact ggatgaattg ttttagctgt cagaccaagt ttactcatat atactttaga
     7741 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     7801 tcatgaccaa aatcccttaa cgtgagtttt cgttctagct gtaacctcga agcgtttcac
     7861 ttgtaacaac gattgagaat ttttgtcata aaattgaaat acttggttcg catttttgtc
     7921 atccgcggtc agccgcaatt ctgacgaact gcccatttag ctggagatga ttgtacatcc
     7981 ttcacgtgaa aatttctcaa gcgctgtgaa caagggttca gattttagat tgaaaggtga
     8041 gccgttgaaa cacgttcttc ttgtcgatga cgacgtcgct atgcggcatc ttattattga
     8101 ataccttacg atccacgcct tcaaagtgac cgcggtagcc gacagcaccc agttcacaag
     8161 agtactctct tccgcgacgg tcgatgtcgt ggttgttgat ctagatttag gtcgtgaaga
     8221 tgggctcgag atcgttcgta atctggcggc aaagtctgat attccaatca taattatcag
     8281 tggcgaccgc cttgaggaga cggataaagt tgttgcactc gagctaggag caagtgattt
     8341 tatcgctaag ccgttcagta tcagagagtt tctagcacgc attcgggttg ccttgcgcgt
     8401 gcgccccaac gttgtccgct ccaaagaccg acggtctttt tgttttactg actggacact
     8461 taatctcagg caacgtcgct tgatgtccga agctggcggt gaggtgaaac ttacggcagg
     8521 tgagttcaat cttctcctcg cgtttttaga gaaaccccgc gacgttctat cgcgcgagca
     8581 acttctcatt gccagtcgag tacgcgacga ggaggtttat gacaggagta tagatgttct
     8641 cattttgagg ctgcgccgca aacttgaggc ggatccgtca agccctcaac tgataaaaac
     8701 agcaagaggt gccggttatt tctttgacgc ggacgtgcag gtttcgcacg gggggacgat
     8761 ggcagcctga gccaattgca tttgcctctt aattatctgg ctcaaagggt gactgaggag
     8821 taagcgatgt gcccatcaca ctgaccacc
//
