LOCUS       sequence-333360-        3126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB4/attB1r entry clone for baculovirus expression of 
            3xFLAG tagged proteins using the polyhedrin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3126)
  AUTHORS   Wall VE, Garvey LA, Mehalko JL, Procter LV, Esposito D
  TITLE     Combinatorial assembly of clone libraries using site-specific 
            recombination.
  JOURNAL   Methods Mol Biol. 2014;1116:193-208. doi: 
            10.1007/978-1-62703-764-8_14.
  PUBMED    24395366
REFERENCE   2  (bases 1 to 3126)
  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: "Methods Mol
            Biol."; date: "2014"; volume: "1116:193-208. doi"; pages: " 
            10.1007/978-1-62703-764-8_14"
COMMENT     Sequence Label: C417-E10: attB4-polyhedrinp>3xFLAG-nostop-attB1r
FEATURES             Location/Qualifiers
     source          1..3126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     42..64
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     56..73
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     57..73
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    129..224
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        290..381
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     306..323
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             385..450
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    451..575
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(647..666)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(648..666)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(671..687)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(671..687)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1112..1903
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      1996..2584
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2485..2504
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2738..2755
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2914..2941)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3033..3119)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
       61 aacgacggcc agtcttaaaa ttcgagctcg gtccgatcgg gcccgcgtta acgctaccat
      121 ggagctccaa ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga
      181 taagcaatgc ttttttataa tgccaacttt gtatagaaaa gttggcgttc gcccaggact
      241 ctagctatag ttctagtggt tggctacgta tactccggaa tattaataga tcatggagat
      301 aattaaaatg ataaccatct cgcaaataaa taagtatttt actgttttcg taacagtttt
      361 gtaataaaaa aacctataaa tatggactat aaggatcatg acggagacta caaagatcac
      421 gatattgatt acaaggatga tgatgataag acaactttgt acaaaaaagt tgaacgagaa
      481 acgtaaaatg atataaatat caatatatta aattagattt tgcataaaaa acagactaca
      541 taatactgta aaacacaaca tatgcagtca ctatgaacca actacttaga tggtattagt
      601 gacctgtaga attcgagctc tagagctgca gggcggccgc gatatcccct atagtgagtc
      661 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
      721 ttacattgca caagatcggt acccggggat cctctagacc agccaggaca gaaatgcctc
      781 gacttcgctg ctacccaagg ttgccgggtg acgcacaccg tggaaacgga tgaaggcacg
      841 aacccagtgg acataagcct gttcggttcg taagctgtaa tgcaagtagc gtatgcgctc
      901 acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt
      961 catggcttgt tatgactgtt tttttggggt acagtctatg cctcgggcat ccaagcagca
     1021 agcgcgttac gccgtgggtc gatgtttgat gttatggagc agcaacgatg ttacgcagca
     1081 gggcagtcgc cctaaaacaa agttaaacat tatgagggaa gcggtgatcg ccgaagtatc
     1141 gactcaacta tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc
     1201 cgtacatttg tacggctccg cagtggatgg cggcctgaag ccacacagtg atattgattt
     1261 gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga tcaacgacct
     1321 tttggaaact tcggcttccc ctggagagag cgagattctc cgcgctgtag aagtcaccat
     1381 tgttgtgcac gacgacatca ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg
     1441 agaatggcag cgcaatgaca ttcttgcagg tatcttcgag ccagccacga tcgacattga
     1501 tctggctatc ttgctgacaa aagcaagaga acatagcgtt gccttggtag gtccagcggc
     1561 ggaggaactc tttgatccgg ttcctgaaca ggatctattt gaggcgctaa atgaaacctt
     1621 aacgctatgg aactcgccgc ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt
     1681 gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga
     1741 ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc atacttgaag ctagacaggc
     1801 ttatcttgga caagaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt
     1861 ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa taaccctcga gccacccatg
     1921 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     1981 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2041 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2101 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     2161 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2221 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2281 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2341 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2401 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2461 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2521 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2581 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2641 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2701 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     2761 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     2821 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg
     2881 taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc
     2941 tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt
     3001 tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc
     3061 gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga
     3121 tgcctg
//
