LOCUS       sequence-107107-        3030 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encodes mature form of murine VEGF-C preceded by honey bee 
            melittin secretion signal; entry vector to Gateway cloning system..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3030)
  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 3030)
  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: HBM-mVEGF-C-HisTag-pENTR
FEATURES             Location/Qualifiers
     source          1..3030
                     /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..751
                     /codon_start=1
                     /product="signal sequence from honeybee melittin"
                     /label=melittin signal sequence
                     /translation="MKFLVNVALVFMVVYISYIYA"
     CDS             1112..1129
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(1155..1254)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1271..1290)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1272..1290)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1295..1311)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1295..1311)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1413..1432)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1424..2233
                     /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(1491..1510)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2380..2968
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2869..2888
                     /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 gaaattctta gtcaacgttg cccttgtttt
      721 tatggtcgta tacatttctt acatctatgc cggatccgca cattataaca cagagatcct
      781 gaaaagtatt gataatgagt ggagaaagac tcaatgcatg ccacgtgagg tgtgtataga
      841 tgtggggaag gagtttggag cagccacaaa caccttcttt aaacctccat gtgtgtccgt
      901 ctacagatgt gggggttgct gcaacagcga ggggctgcag tgcatgaaca ccagcacagg
      961 ttacctcagc aagacgttgt ttgaaattac agtgcctctc tcacaaggcc ccaaaccagt
     1021 cacaatcagt tttgccaatc acacttcctg ccggtgcatg tctaaactgg atgtttacag
     1081 acaagttcat tcaattatta gacgtctcga gcaccaccac caccaccact gaatttaaaa
     1141 gggtgggcgc gccgacccag ctttcttgta caaagttggc attataagaa agcattgctt
     1201 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     1261 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     1321 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
     1381 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     1441 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     1501 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
     1561 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     1621 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     1681 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     1741 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     1801 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     1861 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     1921 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     1981 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     2041 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     2101 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     2161 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     2221 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     2281 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     2341 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     2401 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     2461 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     2521 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     2581 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     2641 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     2701 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     2761 acagcgtgag cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     2821 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     2881 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     2941 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3001 ggccttttgc tggccttttg ctcacatgtt
//
