LOCUS       sequence-242248-        9010 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  binary plant vector for transient expression of GUS (with introns).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9010)
  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 9010)
  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: pSB161 - pL2_pSB90_2x35S::GUS::tMAS
FEATURES             Location/Qualifiers
     source          1..9010
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        445..790
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     733..753
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      2898..3150
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     misc_feature    3214..3238
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3518..4147
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4576..5649
                     /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      5715..5909
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        5971..6075
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6076..6870
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      8195..8783
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8684..8703
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    8919..8943
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 gtgccgatgt atcacctctc gcaagaattc aagcttggag gtcaacatgg tggagcacga
       61 cactctggtc tactccaaaa atgtcaaaga tacagtctca gaagatcaaa gggctattga
      121 gacttttcaa caaaggataa tttcgggaaa cctcctcgga ttccattgcc cagctatctg
      181 tcacttcatc gaaaggacag tagaaaagga aggtggctcc tacaaatgcc atcattgcga
      241 taaaggaaag gctatcattc aagatctctc tgccgacagt ggtcccaaag atggaccccc
      301 acccacgagg agcatcgtgg aaaaagaaga ggttccaacc acgtctacaa agcaagtgga
      361 ttgatgtgat aacatggtgg agcacgacac tctggtctac tccaaaaatg tcaaagatac
      421 agtctcagaa gatcaaaggg ctattgagac ttttcaacaa aggataattt cgggaaacct
      481 cctcggattc cattgcccag ctatctgtca cttcatcgaa aggacagtag aaaaggaagg
      541 tggctcctac aaatgccatc attgcgataa aggaaaggct atcattcaag atctctctgc
      601 cgacagtggt cccaaagatg gacccccacc cacgaggagc atcgtggaaa aagaagaggt
      661 tccaaccacg tctacaaagc aagtggattg atgtgacatc tccactgacg taagggatga
      721 cgcacaatcc cactatcctt cgcaagaccc ttcctctata taaggaagtt catttcattt
      781 ggagaggaca cgctcgagta taagagctca tttttacaac aattaccaac aacaacaaac
      841 aacaaacaac attacaatta catttacaat tatcgataca atgggtcagt cccttatgtt
      901 acgtcctgta gaaaccccaa cccgtgaaat caaaaaactc gacggcctgt gggcattcag
      961 tctggatcgc gaaaactgtg gaattgatca gcgttggtgg gaaagcgcgt tacaagaaag
     1021 ccgggcaatt gctgtgccag gcagttttaa cgatcagttc gccgatgcag atattcgtaa
     1081 ttatgcgggc aacgtctggt atcagcgcga agtctttata ccgaaaggta agtcttactc
     1141 tctctttttt ggtctgtatt tttaattttt tgaagtatac tatttgtact gacgctaata
     1201 atcttttttc aggttgggca ggccagcgta tcgtgctgcg tttcgatgcg gtcactcatt
     1261 acggcaaagt gtgggtcaat aatcaggaag tgatggagca tcagggcggc tatacgccat
     1321 ttgaagccga tgtcacgccg tatgttattg ccgggaaaag tgtacgtatc accgtttgtg
     1381 tgaacaacga actgaactgg cagactatcc cgccgggaat ggtgattacc gacgaaaacg
     1441 gcaagaaaaa gcagtcttac ttccatgatt tctttaacta tgccggaatc catcgcagcg
     1501 taatgctcta caccacgccg aacacctggg tggacgatat caccgtggtg acgcatgtcg
     1561 cgcaagactg taaccacgcg tctgttgact ggcaggtact tcatgcttca acgtgtaact
     1621 taagagatac tgtgtgaaat tttatatttc catacatttg cttgaccttt gctttttgtc
     1681 aatttttttc cccttacagg tggtggccaa tggtgatgtc agcgttgaac tgcgtgatgc
     1741 ggatcaacag gtggttgcaa ctggacaagg cactagcggg actttgcaag tggtgaatcc
     1801 gcacctctgg caaccgggtg aaggttatct ctatgaactg tgcgtcacag ccaaaagcca
     1861 gacagagtgt gatatctacc cgcttcgcgt cggcatccgg tcagtggcag tgaagggcga
     1921 acagttcctg attaaccaca aaccgttcta ctttactggc tttggtcgtc atgaagatgc
     1981 ggacttgcgt ggcaaaggat tcgataacgt gctgatggtg cacgaccacg cattaatgga
     2041 ctggattggg gccaactcct accgtacctc gcattaccct tacgctgaag agatgctcga
     2101 ctgggcagat gaacatggca tcgtggtgat tgatgaaact gctgctgtcg gctttaacct
     2161 ctctttaggc attggtttcg aagcgggcaa caagccgaaa gaactgtaca gcgaagaggc
     2221 agtcaacggg gaaactcagc aagcgcactt acaggcgatt aaagagctga tagcgcgtga
     2281 caaaaaccac ccaagcgtgg tgatgtggag tattgccaac gaaccggata cccgtccgca
     2341 aggtgcacgg gaatatttcg cgccactggc ggaagcaacg cgtaaactcg acccgacgcg
     2401 tccgatcacc tgcgtcaatg taatgttctg cgacgctcac accgatacca tcagcgatct
     2461 ctttgatgtg ctgtgcctga accgttatta cggatggtat gtccaaagcg gcgatttgga
     2521 aacggcagag aaggtactgg aaaaagaact tctggcctgg caggagaaac tgcatcagcc
     2581 gattatcatc accgaatacg gcgtggatac gttagccggg ctgcactcaa tgtacaccga
     2641 catgtggagt gaagagtatc agtgtgcatg gctggatatg tatcaccgcg tctttgatcg
     2701 cgtcagcgcc gtcgtcggtg aacaggtatg gaatttcgcc gattttgcga cctcgcaagg
     2761 catattgcgc gttggcggta acaagaaagg gatcttcact cgcgaccgca aaccgaagtc
     2821 ggcggctttt ctgctgcaaa aacgctggac tggcatgaac ttcggtgaaa aaccgcagca
     2881 gggaggcaaa caatgagctt ggactcccat gttggcaaag gcaaccaaac aaacaatgaa
     2941 tgatccgctc ctgcatatgg ggcggtttga gtatttcaac tgccatttgg gctgaattgt
     3001 agacatgctc ctgtcagaaa ttccgtgatc ttactcaata ttcagtaatc tcggccaata
     3061 tcctaaatgt gcgtggcttt atctgtcttt gtattgtttc atcaattcat gtaacgtttg
     3121 cttttcttat gaattttcaa ataaattatc cgctactaga ggatgcacat gtgaccgagg
     3181 gacacgaagt gatccgttta aactatcagt gtttgacagg atatattggc gggtaaacct
     3241 aagagaaaag agcgtttatt agaataatcg gatatttaaa agggcgtgaa aaggtttatc
     3301 cgttcgtcca tttgtatgtg ccagccgtgc ggctgcatga aatcctggcc ggtttgtctg
     3361 atgccaagct ggcggcctgg ccggccagct tggccgctga agaaaccgag cgccgccgtc
     3421 taaaaaggtg atgtgtattt gagtaaaaca gcttgcgtca tgcggtcgct gcgtatatga
     3481 tgcgatgagt aaataaacaa atacgcaagg ggaacgcatg aaggttatcg ctgtacttaa
     3541 ccagaaaggc gggtcaggca agacgaccat cgcaacccat ctagcccgcg ccctgcaact
     3601 cgccggggcc gatgttctgt tagtcgattc cgatccccag ggcagtgccc gcgattgggc
     3661 ggccgtgcgg gaagatcaac cgctaaccgt tgtcggcatc gaccgcccga cgattgaccg
     3721 cgacgtgaag gccatcggcc ggcgcgactt cgtagtgatc gacggagcgc cccaggcggc
     3781 ggacttggct gtgtccgcga tcaaggcagc cgacttcgtg ctgattccgg tgcagccaag
     3841 cccttacgac atatgggcca ccgccgacct ggtggagctg gttaagcagc gcattgaggt
     3901 cacggatgga aggctacaag cggcctttgt cgtgtcgcgg gcgatcaaag gcacgcgcat
     3961 cggcggtgag gttgccgagg cgctggccgg gtacgagctg cccattcttg agtcccgtat
     4021 cacgcagcgc gtgagctacc caggcactgc cgccgccggc acaaccgttc ttgaatcaga
     4081 acccgagggc gacgctgccc gcgaggtcca ggcgctggcc gctgaaatta aatcaaaact
     4141 catttgagtt aatgaggtaa agagaaaatg agcaaaagca caaacacgct aagtgccggc
     4201 cgtccgagcg cacgcagcag caaggctgca acgttggcca gcctggcaga cacgccagcc
     4261 atgaagcggg tcaactttca gttgccggcg gaggatcaca ccaagctgaa gatgtacgcg
     4321 gtacgccaag gcaagaccat taccgagctg ctatctgaat acatcgcgca gctaccagag
     4381 taaatgagca aatgaataaa tgagtagatg aattttagcg gctaaaggag gcggcatgga
     4441 aaatcaagaa caaccaggca ccgacgccgt ggaatgcccc atgtgtggag gaacgggcgg
     4501 ttggccaggc gtaagcggct gggttgtctg ccggccctgc aatggcactg gaacccccaa
     4561 gcccgaggaa tcggcgtgag cggtcgcaaa ccatccggcc cggtacaaat cggcgcggcg
     4621 ctgggtgatg acctggtgga gaagttgaag gccgcgcagg ccgcccagcg gcaacgcatc
     4681 gaggcagaag cacgccccgg tgaatcgtgg caagcggccg ctgatcgaat ccgcaaagaa
     4741 tcccggcaac cgccggcagc cggtgcgtcg tcgattagga agccgcccaa gggcgacgag
     4801 caaccagatt ttttcgttcc gatgctctat gacgtgggca cccgcgatag tcgcagcatc
     4861 atggacgtgg ccgttttccg tctgtcgaag cgtgaccgac gagctggcga ggtgatccgc
     4921 tacgagcttc cagacgggca cgtagaggtt tccgcagggc cggccggcat ggccagtgtg
     4981 tgggattacg acctggtact gatggcggtt tcccatctaa ccgaatccat gaaccgatac
     5041 cgggaaggga agggagacaa gcccggccgc gtgttccgtc cacacgttgc ggacgtactc
     5101 aagttctgcc ggcgagccga tggcggaaag cagaaagacg acctggtaga aacctgcatt
     5161 cggttaaaca ccacgcacgt tgccatgcag cgtacgaaga aggccaagaa cggccgcctg
     5221 gtgacggtat ccgagggtga agccttgatt agccgctaca agatcgtaaa gagcgaaacc
     5281 gggcggccgg agtacatcga gatcgagcta gctgattgga tgtaccgcga gatcacagaa
     5341 ggcaagaacc cggacgtgct gacggttcac cccgattact ttttgatcga tcccggcatc
     5401 ggccgttttc tctaccgcct ggcacgccgc gccgcaggca aggcagaagc cagatggttg
     5461 ttcaagacga tctacgaacg cagtggcagc gccggagagt tcaagaagtt ctgtttcacc
     5521 gtgcgcaagc tgatcgggtc aaatgacctg ccggagtacg atttgaagga ggaggcgggg
     5581 caggctggcc cgatcctagt catgcgctac cgcaacctga tcgagggcga agcatccgcc
     5641 ggttcctaat gtacggagca gatgctaggg caaattgccc tagcagggga aaaaggtcga
     5701 aaaagcttct ttcctgtgga tagcacgtac attgggaacc caaagccgta cattgggaac
     5761 cggaacccgt acattgggaa cccaaagccg tacattggga accggtcaca catgtaagtg
     5821 actgatataa aagagaaaaa aggcgatttt tccgcctaaa actctttaaa acttattaaa
     5881 actcttaaaa cccgcctggc ctgtgcataa ctgtctggcc agcgcacagc cgaacagctg
     5941 caaaaagcgc ccacatcaag gctccgagtg cgcggaaccc ctatttgttt atttttctaa
     6001 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     6061 tgaaaaagga agagtatggc taaaatgaga atatcaccgg aattgaaaaa actgatcgaa
     6121 aaataccgct gcgtaaaaga tacggaagga atgtctcctg ctaaggtata taagctggtg
     6181 ggagaaaatg aaaacctata tttaaaaatg acggacagcc ggtataaagg gaccacctat
     6241 gatgtggaac gggaaaagga catgatgcta tggctggaag gaaagctgcc tgttccaaag
     6301 gtcctgcact ttgaacggca tgatggctgg agcaatctgc tcatgagtga ggccgatggc
     6361 gtcctttgct cggaagagta tgaagatgaa caaagccctg aaaagattat cgagctgtat
     6421 gcggagtgca tcaggctctt tcactccatc gacatatcgg attgtcccta tacgaatagc
     6481 ttagacagcc gcttagccga attggattac ttactgaata acgatctggc cgatgtggat
     6541 tgcgaaaact gggaagagga cactccattt aaagatccgc gcgagctgta tgatttttta
     6601 aagacggaaa agcccgaaga ggaacttgtc ttttcccacg gcgacctggg agacagcaac
     6661 atctttgtga aagatggcaa agtaagtggc tttattgatc ttgggagaag cggcagggcg
     6721 gacaagtggt atgacattgc cttctgcgtc cggtcgctca gggaggatat cggggaagaa
     6781 cagtatgtcg agctattttt tgacttactg gggatcaagc ctgattggga gaaaataaaa
     6841 tattatattt tactggatga attgttttag ctgtcagacc aagtttactc atatatactt
     6901 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6961 aatctcatga ccaaaatccc ttaacgtgag ttttcgttct agctgtaacc tcgaagcgtt
     7021 tcacttgtaa caacgattga gaatttttgt cataaaattg aaatacttgg ttcgcatttt
     7081 tgtcatccgc ggtcagccgc aattctgacg aactgcccat ttagctggag atgattgtac
     7141 atccttcacg tgaaaatttc tcaagcgctg tgaacaaggg ttcagatttt agattgaaag
     7201 gtgagccgtt gaaacacgtt cttcttgtcg atgacgacgt cgctatgcgg catcttatta
     7261 ttgaatacct tacgatccac gccttcaaag tgaccgcggt agccgacagc acccagttca
     7321 caagagtact ctcttccgcg acggtcgatg tcgtggttgt tgatctagat ttaggtcgtg
     7381 aagatgggct cgagatcgtt cgtaatctgg cggcaaagtc tgatattcca atcataatta
     7441 tcagtggcga ccgccttgag gagacggata aagttgttgc actcgagcta ggagcaagtg
     7501 attttatcgc taagccgttc agtatcagag agtttctagc acgcattcgg gttgccttgc
     7561 gcgtgcgccc caacgttgtc cgctccaaag accgacggtc tttttgtttt actgactgga
     7621 cacttaatct caggcaacgt cgcttgatgt ccgaagctgg cggtgaggtg aaacttacgg
     7681 caggtgagtt caatcttctc ctcgcgtttt tagagaaacc ccgcgacgtt ctatcgcgcg
     7741 agcaacttct cattgccagt cgagtacgcg acgaggaggt ttatgacagg agtatagatg
     7801 ttctcatttt gaggctgcgc cgcaaacttg aggcggatcc gtcaagccct caactgataa
     7861 aaacagcaag aggtgccggt tatttctttg acgcggacgt gcaggtttcg cacgggggga
     7921 cgatggcagc ctgagccaat tgcatttgcc tcttaattat ctggctcaaa gggtgactga
     7981 ggagtaagcg atgtgcccat cacactgacc accaagacgg gggttcgttt tgaagatcac
     8041 ccttcagacc caaggccagt gtgcgttgat cgcggccaca ctgcaaattc acatactgtc
     8101 gataaagggg cgttggacga gaacccattc gtcaggatac tggctcgtga tctcactgag
     8161 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     8221 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     8281 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     8341 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     8401 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     8461 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     8521 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     8581 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     8641 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     8701 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     8761 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctgctcg
     8821 gatctgttgg accggacagt agtcatggtt gatgggctgc ctgtatcgag tggtgatttt
     8881 gtgccgagct gccggtcggg gagctgttgg ctggctggtg gcaggatata ttgtggtgta
     8941 aacaaattga cgcttagaca acttaataac acattgcgga cgtttttaat gtactggggt
     9001 tgaacactct
//
