LOCUS       sequence-333253-        3337 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB2r/attB3 entry clone for construction of C-terminal 
            NanoLuc fusion proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3337)
  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 3337)
  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: C231-E02: attB2r-NanoLuc-attB3
FEATURES             Location/Qualifiers
     source          1..3337
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(533..628)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1039..1830
                     /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      1923..2511
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2412..2431
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2665..2682
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2841..2868)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2960..3046)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3095..3117
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3109..3126
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3110..3126
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(join(3224..3337,1..10))
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 tacaaagtgg gcagcggcgt gtttactctc gaggatttcg tcggagattg gcgccagacc
       61 gccggttaca accttgacca agtgctcgaa cagggaggag tcagctcact gttccagaat
      121 ctcggggtgt ctgtgactcc tatccagcgc atcgtccttt ctggcgaaaa cgggcttaag
      181 atcgacatcc atgtgattat tccctacgag ggactgtcag gcgaccagat ggggcaaatt
      241 gaaaagattt tcaaggtggt gtatccggtc gacgatcacc acttcaaggt gatcctccat
      301 tatgggaccc tggtgattga cggagtgacc ccgaatatga tcgactactt cggtcggcct
      361 tacgaaggca ttgctgtgtt cgacggaaag aaaatcactg tcaccggtac cctctggaat
      421 ggtaacaaaa tcattgacga gagactgatc aaccctgacg gatcacttct gttccgggtc
      481 accatcaatg gggtgaccgg ttggaggctg tgcgagcgca tcctggccta atcaacttta
      541 ttatacaaag ttggcattat aaaaaagcat tgcttatcaa tttgttgcaa cgaacaggtc
      601 actatcagtc aaaataaaat cattatttgg agctccatgg tagcgttaac gcggccgcga
      661 tatcggtacc cggggatcct ctagaccagc caggacagaa atgcctcgac ttcgctgcta
      721 cccaaggttg ccgggtgacg cacaccgtgg aaacggatga aggcacgaac ccagtggaca
      781 taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta tgcgctcacg caactggtcc
      841 agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat
      901 gactgttttt ttggggtaca gtctatgcct cgggcatcca agcagcaagc gcgttacgcc
      961 gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcaggg cagtcgccct
     1021 aaaacaaagt taaacattat gagggaagcg gtgatcgccg aagtatcgac tcaactatca
     1081 gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt acatttgtac
     1141 ggctccgcag tggatggcgg cctgaagcca cacagtgata ttgatttgct ggttacggtg
     1201 accgtaaggc ttgatgaaac aacgcggcga gctttgatca acgacctttt ggaaacttcg
     1261 gcttcccctg gagagagcga gattctccgc gctgtagaag tcaccattgt tgtgcacgac
     1321 gacatcattc cgtggcgtta tccagctaag cgcgaactgc aatttggaga atggcagcgc
     1381 aatgacattc ttgcaggtat cttcgagcca gccacgatcg acattgatct ggctatcttg
     1441 ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt
     1501 gatccggttc ctgaacagga tctatttgag gcgctaaatg aaaccttaac gctatggaac
     1561 tcgccgcccg actgggctgg cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg
     1621 tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag
     1681 cgcctgccgg cccagtatca gcccgtcata cttgaagcta gacaggctta tcttggacaa
     1741 gaagaagatc gcttggcctc gcgcgcagat cagttggaag aatttgtcca ctacgtgaaa
     1801 ggcgagatca ccaaggtagt cggcaaataa ccctcgagcc acccatgacc aaaatccctt
     1861 aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     1921 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     1981 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     2041 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     2101 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     2161 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     2221 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     2281 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     2341 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     2401 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     2461 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     2521 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     2581 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     2641 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     2701 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     2761 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac cgctagccag
     2821 gaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttagtttgat
     2881 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt
     2941 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
     3001 taaaacgaaa ggcccagtct tccgactgag cctttcgttt tatttgatgc ctggcagttc
     3061 cctactctcg cgttaacgct agcatggatg ttttcccagt cacgacgttg taaaacgacg
     3121 gccagtctta aaattcgagc tcggtccgat atccttaagc tcgggccctg cagctctaga
     3181 gctcgaattc tacaggtcac taataccatc taagtagttg gttcatagtg actgcatatg
     3241 ttgtgtttta cagtattatg tagtctgttt tttatgcaaa atctaattta atatattgat
     3301 atttatatca ttttacgttt ctcgttcaac tttcttg
//
