LOCUS       sequence-8633-ex        3023 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  modified pENTR vector for C-terminal ECFP tag fusion with your gene 
            of interest.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3023)
  AUTHORS   Dubin MJ, Bowler C, Benvenuto G
  TITLE     A modified Gateway cloning strategy for overexpressing tagged 
            proteins in plants.
  JOURNAL   Plant Methods. 2008 Jan 22. 4(1):3.
  PUBMED    18211686
REFERENCE   2  (bases 1 to 3023)
  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: "Plant 
            Methods. 2008 Jan 22. 4(1):3."
COMMENT     Sequence Label: pE5c
FEATURES             Location/Qualifiers
     source          1..3023
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      103..189
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     316..339
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    358..457
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(518..520,521..523,524..1237)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(563..584)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(824..843)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1171..1192
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1252..1351)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1463..1482)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1474..2283
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1541..1560)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2373..2961
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2862..2881
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggcttta aaggaaccaa ttcagtcgac
      481 tggatccggt acagatttcg gacatgcggc cgcacctatg gtgagcaagg gcgaggagct
      541 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
      601 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
      661 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctgggg
      721 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
      781 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
      841 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
      901 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacatcag
      961 ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat
     1021 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     1081 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     1141 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     1201 cgggatcact ctcggcatgg acgagctgta caagtaactc gagatatcta gacccagctt
     1261 tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg
     1321 tcactatcag tcaaaataaa atcattattt gccatccagc tgcagctctg gcccgtgtct
     1381 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg
     1441 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg
     1501 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     1561 aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag
     1621 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     1681 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     1741 gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa
     1801 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgtccct gcgccggttg
     1861 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
     1921 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     1981 ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat
     2041 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     2101 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     2161 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     2221 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     2281 taatcagaat tggttaattg gttgtaacat tattcagatt gggccccgtt ccactgagcg
     2341 tcagacccgg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2401 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     2461 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     2521 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2581 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2641 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2701 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     2761 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     2821 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     2881 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     2941 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     3001 tgctggcctt ttgctcacat gtt
//
