LOCUS       sequence-333256-        2898 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB2r/attB3 entry clone for construction of C-terminal 
            Strep2 tagged fusion proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2898)
  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 2898)
  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: C234-E24: attB2r-Strep2-attB3
FEATURES             Location/Qualifiers
     source          1..2898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     30..52
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     44..61
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     45..61
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(124..247)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             256..279
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(356..375)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    complement(400..495)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             873..1664
                     /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      1756..2344
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2245..2264
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2498..2515
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2674..2701)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2793..2879)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tctcgcgtta acgctagcat ggatgttttc ccagtcacga cgttgtaaaa cgacggccag
       61 tcttaaaatt cgagctcggt ccgaggatcc tacaggtcac taataccatc taagtagttg
      121 attcatagtg actggatatg ttgtgtttta cagtattatg tagtctgttt tttatgcaaa
      181 atctaattta atatattgat atttatatca ccttacgttt ctcgttcagc tttcttgtac
      241 aaagtgggcg gatcctggag ccatccgcaa ttcgaaaaag gctaactgaa tagagaaaaa
      301 aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc
      361 atttcagtca gttgctcaat gtacctataa ccagaccgtc aactttatta tacaaagttg
      421 gcattatagg aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa
      481 ataaaatcat tatttgcatg caagcttaga tcctctagac cagccaggac agaaatgcct
      541 cgacttcgct gctacccaag gttgccgggt gacgcacacc gtggaaacgg atgaaggcac
      601 gaacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag cgtatgcgct
      661 cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt
      721 tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc
      781 aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc
      841 agggcagtcg ccctaaaaca aagttaaaca ttatgaggga agcggtgatc gccgaagtat
      901 cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg
      961 ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt
     1021 tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc
     1081 ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca
     1141 ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg
     1201 gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg
     1261 atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg
     1321 cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct
     1381 taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt
     1441 tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg
     1501 actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg
     1561 cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg
     1621 tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg agccacccat
     1681 gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa
     1741 agatcaaaga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     1801 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1861 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     1921 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     1981 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2041 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2101 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2161 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2221 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2281 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2341 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2401 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2461 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     2521 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     2581 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg
     2641 taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc
     2701 tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt
     2761 tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc
     2821 gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga
     2881 tgcctggcag ttccctac
//
