LOCUS       sequence-8581-ex        2564 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for NTAP (N-terminal affinity purification), 
            frame A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2564)
  AUTHORS   Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO, 
            Campisi J, Yaswen P, Cooper PK, Kaufman PD.
  TITLE     A versatile viral system for expression and depletion of proteins in
            mammalian cells.
  JOURNAL   PLoS One. 2009 Aug 6;4(8):e6529.
  PUBMED    19657394
REFERENCE   2  (bases 1 to 2564)
  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: "2009-08-6"; volume: "4(8)"; pages: "e6529"
COMMENT     Sequence Label: pENTR1A-NTAP-A (w322-1)
FEATURES             Location/Qualifiers
     source          1..2564
                     /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             489..566
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             585..698
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     primer_bind     complement(766..782)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(767..783)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    complement(793..892)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1004..1023)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1015..1824
                     /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(1082..1101)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1914..2502
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2403..2422
                     /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 aaggaacccg gtggcggccg
      481 ccaccatgaa gcgacgatgg aaaaagaatt tcatagccgt ctcagcagcc aaccgcttta
      541 agaaaatctc atcctccggg gcacttggaa gcggtagcgg taccatggac gagaagacca
      601 ccggctggcg gggcggccac gtggtggagg gcctggccgg cgagctggag cagctgcggg
      661 ccaggctgga gcaccaccct cagggccagc gggagccctc cggcggctgc aagctgggcg
      721 cccgggcgga tcccccgggc tgcaggaatt cgatatcaag cttatcgata ccgtcgacct
      781 cgagatatct agacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat
      841 caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag
      901 ctgcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata
      961 tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
     1021 catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat
     1081 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat
     1141 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     1201 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     1261 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa
     1321 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     1381 gcagtgtccc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     1441 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     1501 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
     1561 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     1621 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     1681 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     1741 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     1801 ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ttattcagat
     1861 tgggccccgt tccactgagc gtcagacccg gtagaaaaga tcaaaggatc ttcttgagat
     1921 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     1981 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2041 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     2101 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2161 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2221 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2281 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     2341 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     2401 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     2461 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     2521 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtt
//
