LOCUS       sequence-242246-        7531 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  binary plant vector for transient expression of ZCT1 from 
            Catharanthus roseus .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7531)
  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 7531)
  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: pSB153 - pL2_pSB90_2x35S::ZCT1::tMAS
FEATURES             Location/Qualifiers
     source          1..7531
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    99..123
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        635..980
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     923..943
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      1609..1861
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     misc_feature    1925..1949
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             2229..2858
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3287..4360
                     /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      4426..4620
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        4682..4786
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4787..5581
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      6906..7494
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7395..7414
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gatctgttgg accggacagt agtcatggtt gatgggctgc ctgtatcgag tggtgatttt
       61 gtgccgagct gccggtcggg gagctgttgg ctggctggtg gcaggatata ttgtggtgta
      121 aacaaattga cgcttagaca acttaataac acattgcgga cgtttttaat gtactggggt
      181 tgaacactct gtgccgatgt atcacctctc gcaagaattc aagcttggag gtcaacatgg
      241 tggagcacga cactctggtc tactccaaaa atgtcaaaga tacagtctca gaagatcaaa
      301 gggctattga gacttttcaa caaaggataa tttcgggaaa cctcctcgga ttccattgcc
      361 cagctatctg tcacttcatc gaaaggacag tagaaaagga aggtggctcc tacaaatgcc
      421 atcattgcga taaaggaaag gctatcattc aagatctctc tgccgacagt ggtcccaaag
      481 atggaccccc acccacgagg agcatcgtgg aaaaagaaga ggttccaacc acgtctacaa
      541 agcaagtgga ttgatgtgat aacatggtgg agcacgacac tctggtctac tccaaaaatg
      601 tcaaagatac agtctcagaa gatcaaaggg ctattgagac ttttcaacaa aggataattt
      661 cgggaaacct cctcggattc cattgcccag ctatctgtca cttcatcgaa aggacagtag
      721 aaaaggaagg tggctcctac aaatgccatc attgcgataa aggaaaggct atcattcaag
      781 atctctctgc cgacagtggt cccaaagatg gacccccacc cacgaggagc atcgtggaaa
      841 aagaagaggt tccaaccacg tctacaaagc aagtggattg atgtgacatc tccactgacg
      901 taagggatga cgcacaatcc cactatcctt cgcaagaccc ttcctctata taaggaagtt
      961 catttcattt ggagaggaca cgctcgagta taagagctca tttttacaac aattaccaac
     1021 aacaacaaac aacaaacaac attacaatta catttacaat tatcgataca atgggcgtga
     1081 agagattcag agaagaacaa tccgcaaatt gtttgatgct tctttctaaa gtcggattat
     1141 tatcagaaat ggagaaatca gcagtaccgg cgttgaaacc cggcgccggc ggacgaattt
     1201 atgagtgcaa aacttgtaaa aagcaatttc tcacttttca ggctttaggc ggtcaccgag
     1261 cgagtcacaa gaagcttaga ttaatggcgg cagatctact ccatcaatct ttagcggtga
     1321 cgaagccgaa aactcatgct tgttctatat gtggcctcga atttccatta ggtcaggctc
     1381 tcggaggtca tatgagacgc caccgcggcg ccgccttgga tggagagaaa ccggtggtgg
     1441 tgagtgataa acctgttgca aaggcggtac cattcctgat gagatctaat agcagtaaga
     1501 ggatttttgg atttgaagtt gacggattgg atctgaattt gacgcctgag gacaacgatc
     1561 cggacgagcg agacaagttt cctcccttac tggaattttt cgtctaagct tggactccca
     1621 tgttggcaaa ggcaaccaaa caaacaatga atgatccgct cctgcatatg gggcggtttg
     1681 agtatttcaa ctgccatttg ggctgaattg tagacatgct cctgtcagaa attccgtgat
     1741 cttactcaat attcagtaat ctcggccaat atcctaaatg tgcgtggctt tatctgtctt
     1801 tgtattgttt catcaattca tgtaacgttt gcttttctta tgaattttca aataaattat
     1861 ccgctactag aggatgcaca tgtgaccgag ggacacgaag tgatccgttt aaactatcag
     1921 tgtttgacag gatatattgg cgggtaaacc taagagaaaa gagcgtttat tagaataatc
     1981 ggatatttaa aagggcgtga aaaggtttat ccgttcgtcc atttgtatgt gccagccgtg
     2041 cggctgcatg aaatcctggc cggtttgtct gatgccaagc tggcggcctg gccggccagc
     2101 ttggccgctg aagaaaccga gcgccgccgt ctaaaaaggt gatgtgtatt tgagtaaaac
     2161 agcttgcgtc atgcggtcgc tgcgtatatg atgcgatgag taaataaaca aatacgcaag
     2221 gggaacgcat gaaggttatc gctgtactta accagaaagg cgggtcaggc aagacgacca
     2281 tcgcaaccca tctagcccgc gccctgcaac tcgccggggc cgatgttctg ttagtcgatt
     2341 ccgatcccca gggcagtgcc cgcgattggg cggccgtgcg ggaagatcaa ccgctaaccg
     2401 ttgtcggcat cgaccgcccg acgattgacc gcgacgtgaa ggccatcggc cggcgcgact
     2461 tcgtagtgat cgacggagcg ccccaggcgg cggacttggc tgtgtccgcg atcaaggcag
     2521 ccgacttcgt gctgattccg gtgcagccaa gcccttacga catatgggcc accgccgacc
     2581 tggtggagct ggttaagcag cgcattgagg tcacggatgg aaggctacaa gcggcctttg
     2641 tcgtgtcgcg ggcgatcaaa ggcacgcgca tcggcggtga ggttgccgag gcgctggccg
     2701 ggtacgagct gcccattctt gagtcccgta tcacgcagcg cgtgagctac ccaggcactg
     2761 ccgccgccgg cacaaccgtt cttgaatcag aacccgaggg cgacgctgcc cgcgaggtcc
     2821 aggcgctggc cgctgaaatt aaatcaaaac tcatttgagt taatgaggta aagagaaaat
     2881 gagcaaaagc acaaacacgc taagtgccgg ccgtccgagc gcacgcagca gcaaggctgc
     2941 aacgttggcc agcctggcag acacgccagc catgaagcgg gtcaactttc agttgccggc
     3001 ggaggatcac accaagctga agatgtacgc ggtacgccaa ggcaagacca ttaccgagct
     3061 gctatctgaa tacatcgcgc agctaccaga gtaaatgagc aaatgaataa atgagtagat
     3121 gaattttagc ggctaaagga ggcggcatgg aaaatcaaga acaaccaggc accgacgccg
     3181 tggaatgccc catgtgtgga ggaacgggcg gttggccagg cgtaagcggc tgggttgtct
     3241 gccggccctg caatggcact ggaaccccca agcccgagga atcggcgtga gcggtcgcaa
     3301 accatccggc ccggtacaaa tcggcgcggc gctgggtgat gacctggtgg agaagttgaa
     3361 ggccgcgcag gccgcccagc ggcaacgcat cgaggcagaa gcacgccccg gtgaatcgtg
     3421 gcaagcggcc gctgatcgaa tccgcaaaga atcccggcaa ccgccggcag ccggtgcgtc
     3481 gtcgattagg aagccgccca agggcgacga gcaaccagat tttttcgttc cgatgctcta
     3541 tgacgtgggc acccgcgata gtcgcagcat catggacgtg gccgttttcc gtctgtcgaa
     3601 gcgtgaccga cgagctggcg aggtgatccg ctacgagctt ccagacgggc acgtagaggt
     3661 ttccgcaggg ccggccggca tggccagtgt gtgggattac gacctggtac tgatggcggt
     3721 ttcccatcta accgaatcca tgaaccgata ccgggaaggg aagggagaca agcccggccg
     3781 cgtgttccgt ccacacgttg cggacgtact caagttctgc cggcgagccg atggcggaaa
     3841 gcagaaagac gacctggtag aaacctgcat tcggttaaac accacgcacg ttgccatgca
     3901 gcgtacgaag aaggccaaga acggccgcct ggtgacggta tccgagggtg aagccttgat
     3961 tagccgctac aagatcgtaa agagcgaaac cgggcggccg gagtacatcg agatcgagct
     4021 agctgattgg atgtaccgcg agatcacaga aggcaagaac ccggacgtgc tgacggttca
     4081 ccccgattac tttttgatcg atcccggcat cggccgtttt ctctaccgcc tggcacgccg
     4141 cgccgcaggc aaggcagaag ccagatggtt gttcaagacg atctacgaac gcagtggcag
     4201 cgccggagag ttcaagaagt tctgtttcac cgtgcgcaag ctgatcgggt caaatgacct
     4261 gccggagtac gatttgaagg aggaggcggg gcaggctggc ccgatcctag tcatgcgcta
     4321 ccgcaacctg atcgagggcg aagcatccgc cggttcctaa tgtacggagc agatgctagg
     4381 gcaaattgcc ctagcagggg aaaaaggtcg aaaaagcttc tttcctgtgg atagcacgta
     4441 cattgggaac ccaaagccgt acattgggaa ccggaacccg tacattggga acccaaagcc
     4501 gtacattggg aaccggtcac acatgtaagt gactgatata aaagagaaaa aaggcgattt
     4561 ttccgcctaa aactctttaa aacttattaa aactcttaaa acccgcctgg cctgtgcata
     4621 actgtctggc cagcgcacag ccgaacagct gcaaaaagcg cccacatcaa ggctccgagt
     4681 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4741 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatgg ctaaaatgag
     4801 aatatcaccg gaattgaaaa aactgatcga aaaataccgc tgcgtaaaag atacggaagg
     4861 aatgtctcct gctaaggtat ataagctggt gggagaaaat gaaaacctat atttaaaaat
     4921 gacggacagc cggtataaag ggaccaccta tgatgtggaa cgggaaaagg acatgatgct
     4981 atggctggaa ggaaagctgc ctgttccaaa ggtcctgcac tttgaacggc atgatggctg
     5041 gagcaatctg ctcatgagtg aggccgatgg cgtcctttgc tcggaagagt atgaagatga
     5101 acaaagccct gaaaagatta tcgagctgta tgcggagtgc atcaggctct ttcactccat
     5161 cgacatatcg gattgtccct atacgaatag cttagacagc cgcttagccg aattggatta
     5221 cttactgaat aacgatctgg ccgatgtgga ttgcgaaaac tgggaagagg acactccatt
     5281 taaagatccg cgcgagctgt atgatttttt aaagacggaa aagcccgaag aggaacttgt
     5341 cttttcccac ggcgacctgg gagacagcaa catctttgtg aaagatggca aagtaagtgg
     5401 ctttattgat cttgggagaa gcggcagggc ggacaagtgg tatgacattg ccttctgcgt
     5461 ccggtcgctc agggaggata tcggggaaga acagtatgtc gagctatttt ttgacttact
     5521 ggggatcaag cctgattggg agaaaataaa atattatatt ttactggatg aattgtttta
     5581 gctgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     5641 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5701 gttttcgttc tagctgtaac ctcgaagcgt ttcacttgta acaacgattg agaatttttg
     5761 tcataaaatt gaaatacttg gttcgcattt ttgtcatccg cggtcagccg caattctgac
     5821 gaactgccca tttagctgga gatgattgta catccttcac gtgaaaattt ctcaagcgct
     5881 gtgaacaagg gttcagattt tagattgaaa ggtgagccgt tgaaacacgt tcttcttgtc
     5941 gatgacgacg tcgctatgcg gcatcttatt attgaatacc ttacgatcca cgccttcaaa
     6001 gtgaccgcgg tagccgacag cacccagttc acaagagtac tctcttccgc gacggtcgat
     6061 gtcgtggttg ttgatctaga tttaggtcgt gaagatgggc tcgagatcgt tcgtaatctg
     6121 gcggcaaagt ctgatattcc aatcataatt atcagtggcg accgccttga ggagacggat
     6181 aaagttgttg cactcgagct aggagcaagt gattttatcg ctaagccgtt cagtatcaga
     6241 gagtttctag cacgcattcg ggttgccttg cgcgtgcgcc ccaacgttgt ccgctccaaa
     6301 gaccgacggt ctttttgttt tactgactgg acacttaatc tcaggcaacg tcgcttgatg
     6361 tccgaagctg gcggtgaggt gaaacttacg gcaggtgagt tcaatcttct cctcgcgttt
     6421 ttagagaaac cccgcgacgt tctatcgcgc gagcaacttc tcattgccag tcgagtacgc
     6481 gacgaggagg tttatgacag gagtatagat gttctcattt tgaggctgcg ccgcaaactt
     6541 gaggcggatc cgtcaagccc tcaactgata aaaacagcaa gaggtgccgg ttatttcttt
     6601 gacgcggacg tgcaggtttc gcacgggggg acgatggcag cctgagccaa ttgcatttgc
     6661 ctcttaatta tctggctcaa agggtgactg aggagtaagc gatgtgccca tcacactgac
     6721 caccaagacg ggggttcgtt ttgaagatca cccttcagac ccaaggccag tgtgcgttga
     6781 tcgcggccac actgcaaatt cacatactgt cgataaaggg gcgttggacg agaacccatt
     6841 cgtcaggata ctggctcgtg atctcactga gcgtcagacc ccgtagaaaa gatcaaagga
     6901 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6961 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     7021 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     7081 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     7141 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     7201 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     7261 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     7321 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     7381 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     7441 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     7501 agcaacgcgg cctttttacg gttcctgctc g
//
