LOCUS       sequence-330514-        3038 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB2r/attB3 entry clone for construction of C-terminal FLAG
            epitope tagged fusion proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3038)
  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 3038)
  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-E06: attB2r-FLAG-attB3
FEATURES             Location/Qualifiers
     source          1..3038
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..45
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     37..54
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     38..54
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(152..275)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             281..304
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(363..382)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    complement(499..594)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1005..1796
                     /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      1889..2477
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2378..2397
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2631..2648
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2807..2834)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2926..3012)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaaa
       61 attcgagctc ggtccgatat ccttaagctc gggccctgca gctctagagc tcgaattcta
      121 caggtcacta ataccatcta agtagttggt tcatagtgac tgcatatgtt gtgttttaca
      181 gtattatgta gtctgttttt tatgcaaaat ctaatttaat atattgatat ttatatcatt
      241 ttacgtttct cgttcaactt tcttgtacaa agtgggcggc gattataagg acgatgacga
      301 caagtaataa cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt
      361 ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta
      421 cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca
      481 ttcttgcccg cctgtaatca actttattat acaaagttgg cattataaaa aagcattgct
      541 tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttggagct
      601 ccatggtagc gttaacgcgg ccgcgatatc ggtacccggg gatcctctag accagccagg
      661 acagaaatgc ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac
      721 ggatgaaggc acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt
      781 agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg
      841 cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg
      901 catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg
      961 atgttacgca gcagggcagt cgccctaaaa caaagttaaa cattatgagg gaagcggtga
     1021 tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac
     1081 cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca
     1141 gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt
     1201 tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg
     1261 tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg
     1321 aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca
     1381 cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg
     1441 taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc
     1501 taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg
     1561 tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg
     1621 atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg
     1681 aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt
     1741 tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct
     1801 cgagccaccc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag
     1861 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     1921 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     1981 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     2041 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     2101 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     2161 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     2221 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     2281 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     2341 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     2401 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     2461 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     2521 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     2581 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     2641 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     2701 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     2761 caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt
     2821 caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca
     2881 ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag
     2941 gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt
     3001 tcgttttatt tgatgcctgg cagttcccta ctctcgcg
//
