LOCUS       sequence-242247-        7597 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  binary plant vector for transient expression of ORCA3 from 
            Catharanthus roseus .
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: pSB160 - pL2_pSB90_2x35S::ORCA3::tMAS
FEATURES             Location/Qualifiers
     source          1..7597
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        165..510
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     453..473
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      1205..1457
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     misc_feature    1521..1545
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             1825..2454
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2883..3956
                     /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      4022..4216
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        4278..4382
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4383..5177
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      6502..7090
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6991..7010
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7226..7250
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 ggtcccaaag atggaccccc acccacgagg agcatcgtgg aaaaagaaga ggttccaacc
       61 acgtctacaa agcaagtgga ttgatgtgat aacatggtgg agcacgacac tctggtctac
      121 tccaaaaatg tcaaagatac agtctcagaa gatcaaaggg ctattgagac ttttcaacaa
      181 aggataattt cgggaaacct cctcggattc cattgcccag ctatctgtca cttcatcgaa
      241 aggacagtag aaaaggaagg tggctcctac aaatgccatc attgcgataa aggaaaggct
      301 atcattcaag atctctctgc cgacagtggt cccaaagatg gacccccacc cacgaggagc
      361 atcgtggaaa aagaagaggt tccaaccacg tctacaaagc aagtggattg atgtgacatc
      421 tccactgacg taagggatga cgcacaatcc cactatcctt cgcaagaccc ttcctctata
      481 taaggaagtt catttcattt ggagaggaca cgctcgagta taagagctca tttttacaac
      541 aattaccaac aacaacaaac aacaaacaac attacaatta catttacaat tatcgataca
      601 atgtccgaag aaatcatttc cgtctcagat cgatttcttc tttccttaat cgaagaacat
      661 cttctcagcg ataattctga tgattccagc tcggaattga cttctacaga ggaaaattgg
      721 gaagaaattt ttgcagattt tctaaattgg tcgggatccg aaatacagaa acgcggtagc
      781 ccgagttccg aaagctgtca atcgaattca atggcggaaa gctgtcagga ggattctgtt
      841 gtgggaaccc cgccagaagc ggcggccgga ggaggttgtt cgaaggattg gaaccggtat
      901 aagggcgtta gacggcggcc gtgggggaag ttcgcggcgg agataaggga tccgaaaaag
      961 aaaggatcca ggatttggtt gggtacatac gagacacctg aggatgcagc attggcttat
     1021 gatgcagccg cgtttaatat gcgtggagct aaagctaggc ttaattttcc tcatttgatt
     1081 ggttcgaata tttccggacc cgttagagta aacccgagaa aacgtttccc tgcggagcct
     1141 tctacgacgt cgtcgtcatc ttcgtctgaa aatagtggag gaaggaagaa gagacgatat
     1201 taagcttgga ctcccatgtt ggcaaaggca accaaacaaa caatgaatga tccgctcctg
     1261 catatggggc ggtttgagta tttcaactgc catttgggct gaattgtaga catgctcctg
     1321 tcagaaattc cgtgatctta ctcaatattc agtaatctcg gccaatatcc taaatgtgcg
     1381 tggctttatc tgtctttgta ttgtttcatc aattcatgta acgtttgctt ttcttatgaa
     1441 ttttcaaata aattatccgc tactagagga tgcacatgtg accgagggac acgaagtgat
     1501 ccgtttaaac tatcagtgtt tgacaggata tattggcggg taaacctaag agaaaagagc
     1561 gtttattaga ataatcggat atttaaaagg gcgtgaaaag gtttatccgt tcgtccattt
     1621 gtatgtgcca gccgtgcggc tgcatgaaat cctggccggt ttgtctgatg ccaagctggc
     1681 ggcctggccg gccagcttgg ccgctgaaga aaccgagcgc cgccgtctaa aaaggtgatg
     1741 tgtatttgag taaaacagct tgcgtcatgc ggtcgctgcg tatatgatgc gatgagtaaa
     1801 taaacaaata cgcaagggga acgcatgaag gttatcgctg tacttaacca gaaaggcggg
     1861 tcaggcaaga cgaccatcgc aacccatcta gcccgcgccc tgcaactcgc cggggccgat
     1921 gttctgttag tcgattccga tccccagggc agtgcccgcg attgggcggc cgtgcgggaa
     1981 gatcaaccgc taaccgttgt cggcatcgac cgcccgacga ttgaccgcga cgtgaaggcc
     2041 atcggccggc gcgacttcgt agtgatcgac ggagcgcccc aggcggcgga cttggctgtg
     2101 tccgcgatca aggcagccga cttcgtgctg attccggtgc agccaagccc ttacgacata
     2161 tgggccaccg ccgacctggt ggagctggtt aagcagcgca ttgaggtcac ggatggaagg
     2221 ctacaagcgg cctttgtcgt gtcgcgggcg atcaaaggca cgcgcatcgg cggtgaggtt
     2281 gccgaggcgc tggccgggta cgagctgccc attcttgagt cccgtatcac gcagcgcgtg
     2341 agctacccag gcactgccgc cgccggcaca accgttcttg aatcagaacc cgagggcgac
     2401 gctgcccgcg aggtccaggc gctggccgct gaaattaaat caaaactcat ttgagttaat
     2461 gaggtaaaga gaaaatgagc aaaagcacaa acacgctaag tgccggccgt ccgagcgcac
     2521 gcagcagcaa ggctgcaacg ttggccagcc tggcagacac gccagccatg aagcgggtca
     2581 actttcagtt gccggcggag gatcacacca agctgaagat gtacgcggta cgccaaggca
     2641 agaccattac cgagctgcta tctgaataca tcgcgcagct accagagtaa atgagcaaat
     2701 gaataaatga gtagatgaat tttagcggct aaaggaggcg gcatggaaaa tcaagaacaa
     2761 ccaggcaccg acgccgtgga atgccccatg tgtggaggaa cgggcggttg gccaggcgta
     2821 agcggctggg ttgtctgccg gccctgcaat ggcactggaa cccccaagcc cgaggaatcg
     2881 gcgtgagcgg tcgcaaacca tccggcccgg tacaaatcgg cgcggcgctg ggtgatgacc
     2941 tggtggagaa gttgaaggcc gcgcaggccg cccagcggca acgcatcgag gcagaagcac
     3001 gccccggtga atcgtggcaa gcggccgctg atcgaatccg caaagaatcc cggcaaccgc
     3061 cggcagccgg tgcgtcgtcg attaggaagc cgcccaaggg cgacgagcaa ccagattttt
     3121 tcgttccgat gctctatgac gtgggcaccc gcgatagtcg cagcatcatg gacgtggccg
     3181 ttttccgtct gtcgaagcgt gaccgacgag ctggcgaggt gatccgctac gagcttccag
     3241 acgggcacgt agaggtttcc gcagggccgg ccggcatggc cagtgtgtgg gattacgacc
     3301 tggtactgat ggcggtttcc catctaaccg aatccatgaa ccgataccgg gaagggaagg
     3361 gagacaagcc cggccgcgtg ttccgtccac acgttgcgga cgtactcaag ttctgccggc
     3421 gagccgatgg cggaaagcag aaagacgacc tggtagaaac ctgcattcgg ttaaacacca
     3481 cgcacgttgc catgcagcgt acgaagaagg ccaagaacgg ccgcctggtg acggtatccg
     3541 agggtgaagc cttgattagc cgctacaaga tcgtaaagag cgaaaccggg cggccggagt
     3601 acatcgagat cgagctagct gattggatgt accgcgagat cacagaaggc aagaacccgg
     3661 acgtgctgac ggttcacccc gattactttt tgatcgatcc cggcatcggc cgttttctct
     3721 accgcctggc acgccgcgcc gcaggcaagg cagaagccag atggttgttc aagacgatct
     3781 acgaacgcag tggcagcgcc ggagagttca agaagttctg tttcaccgtg cgcaagctga
     3841 tcgggtcaaa tgacctgccg gagtacgatt tgaaggagga ggcggggcag gctggcccga
     3901 tcctagtcat gcgctaccgc aacctgatcg agggcgaagc atccgccggt tcctaatgta
     3961 cggagcagat gctagggcaa attgccctag caggggaaaa aggtcgaaaa agcttctttc
     4021 ctgtggatag cacgtacatt gggaacccaa agccgtacat tgggaaccgg aacccgtaca
     4081 ttgggaaccc aaagccgtac attgggaacc ggtcacacat gtaagtgact gatataaaag
     4141 agaaaaaagg cgatttttcc gcctaaaact ctttaaaact tattaaaact cttaaaaccc
     4201 gcctggcctg tgcataactg tctggccagc gcacagccga acagctgcaa aaagcgccca
     4261 catcaaggct ccgagtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4321 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4381 gtatggctaa aatgagaata tcaccggaat tgaaaaaact gatcgaaaaa taccgctgcg
     4441 taaaagatac ggaaggaatg tctcctgcta aggtatataa gctggtggga gaaaatgaaa
     4501 acctatattt aaaaatgacg gacagccggt ataaagggac cacctatgat gtggaacggg
     4561 aaaaggacat gatgctatgg ctggaaggaa agctgcctgt tccaaaggtc ctgcactttg
     4621 aacggcatga tggctggagc aatctgctca tgagtgaggc cgatggcgtc ctttgctcgg
     4681 aagagtatga agatgaacaa agccctgaaa agattatcga gctgtatgcg gagtgcatca
     4741 ggctctttca ctccatcgac atatcggatt gtccctatac gaatagctta gacagccgct
     4801 tagccgaatt ggattactta ctgaataacg atctggccga tgtggattgc gaaaactggg
     4861 aagaggacac tccatttaaa gatccgcgcg agctgtatga ttttttaaag acggaaaagc
     4921 ccgaagagga acttgtcttt tcccacggcg acctgggaga cagcaacatc tttgtgaaag
     4981 atggcaaagt aagtggcttt attgatcttg ggagaagcgg cagggcggac aagtggtatg
     5041 acattgcctt ctgcgtccgg tcgctcaggg aggatatcgg ggaagaacag tatgtcgagc
     5101 tattttttga cttactgggg atcaagcctg attgggagaa aataaaatat tatattttac
     5161 tggatgaatt gttttagctg tcagaccaag tttactcata tatactttag attgatttaa
     5221 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5281 aaatccctta acgtgagttt tcgttctagc tgtaacctcg aagcgtttca cttgtaacaa
     5341 cgattgagaa tttttgtcat aaaattgaaa tacttggttc gcatttttgt catccgcggt
     5401 cagccgcaat tctgacgaac tgcccattta gctggagatg attgtacatc cttcacgtga
     5461 aaatttctca agcgctgtga acaagggttc agattttaga ttgaaaggtg agccgttgaa
     5521 acacgttctt cttgtcgatg acgacgtcgc tatgcggcat cttattattg aataccttac
     5581 gatccacgcc ttcaaagtga ccgcggtagc cgacagcacc cagttcacaa gagtactctc
     5641 ttccgcgacg gtcgatgtcg tggttgttga tctagattta ggtcgtgaag atgggctcga
     5701 gatcgttcgt aatctggcgg caaagtctga tattccaatc ataattatca gtggcgaccg
     5761 ccttgaggag acggataaag ttgttgcact cgagctagga gcaagtgatt ttatcgctaa
     5821 gccgttcagt atcagagagt ttctagcacg cattcgggtt gccttgcgcg tgcgccccaa
     5881 cgttgtccgc tccaaagacc gacggtcttt ttgttttact gactggacac ttaatctcag
     5941 gcaacgtcgc ttgatgtccg aagctggcgg tgaggtgaaa cttacggcag gtgagttcaa
     6001 tcttctcctc gcgtttttag agaaaccccg cgacgttcta tcgcgcgagc aacttctcat
     6061 tgccagtcga gtacgcgacg aggaggttta tgacaggagt atagatgttc tcattttgag
     6121 gctgcgccgc aaacttgagg cggatccgtc aagccctcaa ctgataaaaa cagcaagagg
     6181 tgccggttat ttctttgacg cggacgtgca ggtttcgcac ggggggacga tggcagcctg
     6241 agccaattgc atttgcctct taattatctg gctcaaaggg tgactgagga gtaagcgatg
     6301 tgcccatcac actgaccacc aagacggggg ttcgttttga agatcaccct tcagacccaa
     6361 ggccagtgtg cgttgatcgc ggccacactg caaattcaca tactgtcgat aaaggggcgt
     6421 tggacgagaa cccattcgtc aggatactgg ctcgtgatct cactgagcgt cagaccccgt
     6481 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6541 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6601 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6661 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6721 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6781 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6841 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6901 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6961 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     7021 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     7081 cctatggaaa aacgccagca acgcggcctt tttacggttc ctgctcggat ctgttggacc
     7141 ggacagtagt catggttgat gggctgcctg tatcgagtgg tgattttgtg ccgagctgcc
     7201 ggtcggggag ctgttggctg gctggtggca ggatatattg tggtgtaaac aaattgacgc
     7261 ttagacaact taataacaca ttgcggacgt ttttaatgta ctggggttga acactctgtg
     7321 ccgatgtatc acctctcgca agaattcaag cttggaggtc aacatggtgg agcacgacac
     7381 tctggtctac tccaaaaatg tcaaagatac agtctcagaa gatcaaaggg ctattgagac
     7441 ttttcaacaa aggataattt cgggaaacct cctcggattc cattgcccag ctatctgtca
     7501 cttcatcgaa aggacagtag aaaaggaagg tggctcctac aaatgccatc attgcgataa
     7561 aggaaaggct atcattcaag atctctctgc cgacagt
//
