LOCUS       sequence-107106-        3027 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encodes mature form of murine VEGF-C preceded by baculoviral
            gp64 secretion signal; entry vector to Gateway cloning system..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3027)
  AUTHORS   Klaus T, Kulesza M, Bzowska M, Wyroba B, Kilarski WW, Bereta J
  TITLE     Overcoming inefficient secretion of recombinant VEGF-C in 
            baculovirus expression vector system by simple purification of the 
            protein from cell lysate.
  JOURNAL   Protein Expr Purif. 2015 Mar 7. pii: S1046-5928(15)00040-6. doi: 
            10.1016/j.pep.2015.03.001.
  PUBMED    25758709
REFERENCE   2  (bases 1 to 3027)
  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: "Protein 
            Expr Purif. 2015 Mar 7. pii: S1046-5928(15)00040-6. doi: 
            10.1016/j.pep.2015.03.001."
COMMENT     Sequence Label: gp64-mVEGF-C-HisTag-pENTR
FEATURES             Location/Qualifiers
     source          1..3027
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     sig_peptide     689..748
                     /codon_start=1
                     /gene="Autographa californica gp64"
                     /product="signal sequence from the baculovirus envelope 
                     glycoprotein GP64"
                     /label=GP64 signal sequence
                     /translation="MVSAIVLYVLLAAAAHSAFA"
     CDS             1109..1126
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(1152..1251)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1268..1287)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1269..1287)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1292..1308)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1292..1308)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1410..1429)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1421..2230
                     /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
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1488..1507)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2377..2965
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2866..2885
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc gcggccgccc ccttcaccat ggtaagcgct attgttttat atgtgctttt
      721 ggcggcggcg gcgcattctg cctttgcggg atccgcacat tataacacag agatcctgaa
      781 aagtattgat aatgagtgga gaaagactca atgcatgcca cgtgaggtgt gtatagatgt
      841 ggggaaggag tttggagcag ccacaaacac cttctttaaa cctccatgtg tgtccgtcta
      901 cagatgtggg ggttgctgca acagcgaggg gctgcagtgc atgaacacca gcacaggtta
      961 cctcagcaag acgttgtttg aaattacagt gcctctctca caaggcccca aaccagtcac
     1021 aatcagtttt gccaatcaca cttcctgccg gtgcatgtct aaactggatg tttacagaca
     1081 agttcattca attattagac gtctcgagca ccaccaccac caccactgaa tttaaaaggg
     1141 tgggcgcgcc gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     1201 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     1261 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     1321 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     1381 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     1441 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     1501 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     1561 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     1621 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     1681 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     1741 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     1801 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     1861 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     1921 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     1981 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     2041 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     2101 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     2161 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     2221 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     2281 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     2341 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2401 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2461 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2521 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2581 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2641 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     2701 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     2761 gcgtgagcat tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     2821 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     2881 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     2941 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3001 cttttgctgg ccttttgctc acatgtt
//
