LOCUS       sequence-212459-        7776 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plant Gene expression Vector pSIM24 containing eGFP reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7776)
  AUTHORS   Sahoo DK, Dey N, Maiti IB
  TITLE     pSiM24 is a novel versatile gene expression vector for transient 
            assays as well as stable expression of foreign genes in plants.
  JOURNAL   PLoS One. 2014 Jun 4;9(6):e98988. doi: 10.1371/journal.pone.0098988.
            eCollection 2014.
  PUBMED    24897541
REFERENCE   2  (bases 1 to 7776)
  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: "PLoS One.";
            date: "2014-06-4"; volume: "9(6):e98988. doi"; pages: " 
            10.1371/journal.pone.0098988. eCollection 2014"
COMMENT     Sequence Label: pSiM24-eGFP
FEATURES             Location/Qualifiers
     source          1..7776
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    67..91
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             join(877..879,880..882,883..1596)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(877..879,880..882,883..1593)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(922..943)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1183..1202)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1530..1551
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      1614..2255
                     /label=E9 terminator
                     /note="terminator and polyadenylation signal from the pea 
                     rbcS-E9 gene"
     promoter        complement(3335..3514)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    3749..3773
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(3888..4476)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3968..3987)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4647..5438,5439..5507))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFCVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5270..5289
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5508..5612)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5653..6263
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      5740..6271
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 ggcgcgcggt acccagtaca ttaaaaacgt ccgcaatgtg ttattaagtt gtctaagcgt
       61 caatttgttt acaccacaat atatcctgcc accagccagc caacagctcc ccgaccggca
      121 gctcggcaca aaatcaccac tcgatacagg cagcccatca gtccactaga ccagtacctt
      181 ggaggcctgt gcgcatccca gggtgggtcc cgtatacccc ggggaattcg tcaacttcgt
      241 ccacagacat caacatctta tcgtcctttg aagataagat aataatgttg aagataagag
      301 tgggagcccc cactaaaaca ttgctttgtc aaaagctaaa aaagatgatg cccgacagcc
      361 acttgtgtga agcatgagaa gccggtccct ccactaagaa aattagtgaa gcatcttcca
      421 gtggtccctc cactcacagc tcaatcagtg agcaacagga cgaaggaaat gacgtaagcc
      481 atgacgtcta atcccaactt cgtccacaga catcaacatc ttatcgtcct ttgaagataa
      541 gataataatg ttgaagataa gagtgggagc caccactaaa acattgcttt gtcaaaagct
      601 aaaaaagatg atgcccgaca gccacttgtg tgaagcatga gaagccggtc cctccactaa
      661 gaaaattagt gaagcatctt ccagtggtcc ctccactcac agctcaatca gtgagcaaca
      721 ggacgaagga aatgacgtaa gccatgacgt ctaatcccac aagaatttcc ttatataagg
      781 aacacaaatc agaaggaaga gatcaatcga aatcaaaatc ggaatcgaaa tcaaaatcgg
      841 aatcgaaatc tctcatctaa gcttggatcc ctcgagatgg tgagcaaggg cgaggagctg
      901 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
      961 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     1021 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     1081 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     1141 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     1201 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     1261 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     1321 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     1381 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     1441 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     1501 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     1561 gggatcactc tcggcatgga cgagctgtac aagtaagagc tcctgcagtc tagagctttc
     1621 gttcgtatca tcggtttcga caacgttcgt caagttcaat gcatcagttt cattgcgcac
     1681 acaccagaat cctactgagt ttgagtatta tggcattggg aaaactgttt ttcttgtacc
     1741 atttgttgtg cttgtaattt actgtgtttt ttattcggtt ttcgctatcg aactgtgaaa
     1801 tggaaatgga tggagaagag ttaatgaatg atatggtcct tttgttcatt ctcaaattaa
     1861 tattatttgt tttttctctt atttgttgtg tgttgaattt gaaattataa gagatatgca
     1921 aacattttgt tttgagtaaa aatgtgtcaa atcgtggcct ctaatgaccg aagttaatat
     1981 gaggagtaaa acacttgtag ttgtaccatt atgcttattc actaggcaac aaatatattt
     2041 tcagacctag aaaagctgca aatgttactg aatacaagta tgtcctcttg tgttttagac
     2101 atttatgaac tttcctttat gtaattttcc agaatccttg tcagattcta atcattgctt
     2161 tataattata gttatactca tggatttgta gttgagtatg aaaatatttt ttaatgcatt
     2221 ttatgacttg ccaattgatt gacaacatgc atcaatcgat actagtctag taacatagat
     2281 gacaccgcga gcgataattt atcctagttt gcgagctata ttttgttttc tatcgcgtat
     2341 taaatgtata attgcgggac tctaatcata aaaacccatc tcataaataa cgtcatgcat
     2401 tacatgttaa ttattacatg cttaacgtaa ttcaacagaa attatatgat aatcatcgca
     2461 agaccggcaa caggattcaa tcttaagaaa ctttattgcc aaatgtttga acgattcaga
     2521 agaactcatc gaggagcctg tagaaagcaa tcctttggga atctggagca gcaattccgt
     2581 aaagcacaag gaatctatca gcccactctc caccaagttc ttcagcaata tccctagtag
     2641 cgagagcaat atcctgatac ctatcagcaa ctccaagcct tccgcaatca atgaatccag
     2701 aaaaccttcc gttctccacc ataatgtttg gaaggcaagc atcaccatga gtaacaacaa
     2761 gatcctcacc atctggcatt ctagccttga gcctagcaaa aagttcagct ggagccaatc
     2821 cttgatgttc ctcatcaaga tcatcctgat ccacaagtcc agcttccatc ctagttctag
     2881 ccctttcaat tctgtgctta gcttggtgat caaatgggca agtagcagga tcaagagtgt
     2941 gaagccttct catagcatca gccataatgg aaaccttctc agctggagca aggtgtgaag
     3001 aaagcaaatc ttgtcctgga acctctccca aaagaagcca atctcttcca gcctcagtaa
     3061 caacatcaag aacagcagcg catggaacac cagtagtagc aagccaagaa agtctagcag
     3121 cctcatcttg aagctcatta agagcaccag aaagatcagt cttaacgaaa agaactggcc
     3181 ttccctgagc agaaagccta aaaacagcag catcagagca tccaatagtc tgttgagccc
     3241 aatcgtatcc aaaaagcctc tcaacccaag cagcaggact tccagcatga agtccatctt
     3301 gctcaattcc catttgagag tgaatatgag actctaattg gataccgagg ggaatttatg
     3361 gaacgtcagt ggagcatttt tgacaagaaa tatttgctag ctgatagtga ccttaggcga
     3421 cttttgaacg cgcaataatg gtttctgacg tatgtgctta gctcattaaa ctccagaaac
     3481 ccgcggctca gtggctcctt caacgttgcg gttctgtcag ttccaaacgt aaaacggctt
     3541 gtcccgcgtc atcggcgggg gtcataacgt gactccctta attctccgct catgatcaga
     3601 tctggtgacc cctatgtaag gggccggcca tttaaattcg cgattaatta acctgtggtt
     3661 ggcatgcaca tacaaatgga cgaacggata aaccttttca cgccctttta aatatccgat
     3721 tattctaata aacgctcttt tctcttaggt ttacccgcca atatatcctg tcaaacactg
     3781 atagtttaaa ctgaaggcgg gaaacgacaa tctgatatcg cgcgcggtac ctgagcaaaa
     3841 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3901 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3961 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4021 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4081 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4141 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4201 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4261 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4321 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4381 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4441 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4501 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4561 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4621 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4681 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4741 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga accacgctca
     4801 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4861 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4921 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4981 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5041 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5101 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5161 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5221 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5281 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5341 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5401 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5461 gccgcaaaaa agggaataag ggcgacacag aaatgttgaa tactcatact cttccttttt
     5521 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5581 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacgggcc
     5641 ccgctcaccg ggctggttgc cctcgccgct gggctggcgg ccgtctatgg ccctgcaaac
     5701 gcgccagaaa cgccgtcgaa gccgtgtgcg agacaccgcg gccgccggcg ttgtggatac
     5761 ctcgcggaaa acttggccct cactgacaga tgaggggcgg acgttgacac ttgaggggcc
     5821 gactcacccg gcgcggcgtt gacagatgag gggcaggctc gatttcggcc ggcgacgtgg
     5881 agctggccag cctcgcaaat cggcgaaaac gcctgatttt acgcgagttt cccacagatg
     5941 atgtggacaa gcctggggat aagtgccctg cggtattgac acttgagggg cgcgactact
     6001 gacagatgag gggcgcgatc cttgacactt gaggggcaga gtgctgacag atgaggggcg
     6061 cacctattga catttgaggg gctgtccaca ggcagaaaat ccagcatttg caagggtttc
     6121 cgcccgtttt tcggccaccg ctaacctgtc ttttaacctg cttttaaacc aatatttata
     6181 aaccttgttt ttaaccaggg ctgcgccctg tgcgcgtgac cgcgcacgcc gaaggggggt
     6241 gccccccctt ctcgaaccct cccggcccgc tgtcgactac ctagatgtgg cgcaacgatg
     6301 ccggcgacaa gcaggagcgc accgacttct tccgcatcaa gtgttttggc tctcaggccg
     6361 aggcccacgg caagtatttg ggcaaggggt cgctggtatt cgtgcagggc aagattcgga
     6421 ataccaagta cgagaaggac ggccagacgg tctacgggac cgacttcatt gccgataagg
     6481 tggattatct ggacaccaag gcaccaggcg ggtcaaatca ggaataaggg cacattgccc
     6541 cggcgtgagt cggggcaatc ccgcaaggag ggtgaatgaa tcgcaccttc gatcgcaagg
     6601 cgtatcgcca ggaactgatc gacgccggct tctcggccga agatgccgaa acgatcgcct
     6661 cgcgcaccgt catgcgtgcc ccgcgcgaaa ccttccagtc ggtcggctcg atggtccagc
     6721 aggccaccgc caagatcgaa cgcgattcgg tccagctggc cccgccggcc cttccggccc
     6781 cgtcggccgc cgtcgaacgc tcgcgccggc tggaacagga agccgccggc ctggccaagt
     6841 cgatgaccat cgacacccgc ggcaccatga ccaccaagaa gcgcaagacc gccggcgagg
     6901 atctggccaa gcaggtcagc gaagccaagc aggccgccct gctgaagcat accaagcagc
     6961 agatcaagga aatgcagctg tcgctgttcg acatcgcccc gtggccggat accatgcggg
     7021 ccatgccgaa tgataccgcc cgctcggccc tgttcaccac ccgcaataag aagatcccgc
     7081 gcgaagccct ccagaataag gtcatcttcc acgtcaacaa ggatgtcaag atcacctata
     7141 ccggcgtcga actgcgggcc gatgatgatg aactggtctg gcagcaggtc ctggaatatg
     7201 ccaagcgcac cccgatcggc gaaccgatca ccttcacctt ctatgaactg tgccaggatc
     7261 tgggctggtc gatcaatggc cgctattata ccaaggccga agaatgcctg tcgcgcctcc
     7321 aggcgaccgc gatgggcttc acctcggatc gcgtcggcca tctggaatcg gtcagcctgc
     7381 tgcaccgctt ccgcgtcctg gatcgcggca agaagacctc gcgctgccag gtcctgatcg
     7441 acgaagaaat cgtcgtcctg ttcgccggcg atcattatac caagttcatc tgggagaagt
     7501 atcgcaagct gtcgccgacc gcccgccgca tgttcgatta tttctcgtcg caccgcgaac
     7561 cgtatccgct gaagctggaa accttccgcc tgatgtgcgg ctcggattcg acccgcgtca
     7621 agaagtggcg cgaacaggtc ggcgaagcct gcgaagaact gcgcggctcg ggcctggtcg
     7681 aacatgcctg ggtcaatgat gatctggtcc attgcaagcg ctagggcctt gtggggtcag
     7741 ttccggctgg gggttcagcc agcgctttac gatatc
//
