LOCUS       sequence-330932-        2993 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB5r/attB1r entry clone for generation of N-terminal 
            Strep2-tagged fusion proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2993)
  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 2993)
  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: C514-E42: attB5r-RU5u-Strep2-nostop-attB1r
FEATURES             Location/Qualifiers
     source          1..2993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(44..167)
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     LTR             171..439
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             452..475
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    479..603
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(675..694)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(676..694)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(699..715)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(699..715)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1140..1931
                     /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      2024..2612
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2513..2532
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2747..2774)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2866..2952)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 agctcgaatt ctacagtcac taataccatc taagtagttg attcatagtg actgcatatg
       61 ttgtgtttta cagtattatg tagtctgttt tttatgcaaa atctaattta atatattgat
      121 atttatatca ttttacgttt ctcgttcaac ttttgtatac aaagtggcga ggggctcgca
      181 tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg
      241 ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa
      301 agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc
      361 ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg
      421 ttctgcgccg ttacagatca ccatgggatc ctggagccat ccgcaattcg aaaaaggcac
      481 aactttgtac aaaaaagttg aacgagaaac gtaaaatgat ataaatatca atatattaaa
      541 ttagattttg cataaaaaac agactacata atactgtaaa acacaacata tgcagtcact
      601 atgaaccaac tacttagatg gtattagtga cctgtagaat tcgagctcta gagctgcagg
      661 gcggccgcga tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc
      721 tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agatcggtac ccggggatcc
      781 tctagaccag ccaggacaga aatgcctcga cttcgctgct acccaaggtt gccgggtgac
      841 gcacaccgtg gaaacggatg aaggcacgaa cccagtggac ataagcctgt tcggttcgta
      901 agctgtaatg caagtagcgt atgcgctcac gcaactggtc cagaaccttg accgaacgca
      961 gcggtggtaa cggcgcagtg gcggttttca tggcttgtta tgactgtttt tttggggtac
     1021 agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt
     1081 tatggagcag caacgatgtt acgcagcagg gcagtcgccc taaaacaaag ttaaacatta
     1141 tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg
     1201 agcgccatct cgaaccgacg ttgctggccg tacatttgta cggctccgca gtggatggcg
     1261 gcctgaagcc acacagtgat attgatttgc tggttacggt gaccgtaagg cttgatgaaa
     1321 caacgcggcg agctttgatc aacgaccttt tggaaacttc ggcttcccct ggagagagcg
     1381 agattctccg cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt
     1441 atccagctaa gcgcgaactg caatttggag aatggcagcg caatgacatt cttgcaggta
     1501 tcttcgagcc agccacgatc gacattgatc tggctatctt gctgacaaaa gcaagagaac
     1561 atagcgttgc cttggtaggt ccagcggcgg aggaactctt tgatccggtt cctgaacagg
     1621 atctatttga ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc gactgggctg
     1681 gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg gtacagcgca gtaaccggca
     1741 aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga gcgcctgccg gcccagtatc
     1801 agcccgtcat acttgaagct agacaggctt atcttggaca agaagaagat cgcttggcct
     1861 cgcgcgcaga tcagttggaa gaatttgtcc actacgtgaa aggcgagatc accaaggtag
     1921 tcggcaaata accctcgagc cacccatgac caaaatccct taacgtgagt tacgcgtcgt
     1981 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     2041 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     2101 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     2161 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     2221 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     2281 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     2341 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     2401 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     2461 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     2521 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     2581 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     2641 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     2701 tggataaccg tattaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt
     2761 caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca
     2821 ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag
     2881 gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt
     2941 tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctacca tgg
//
