LOCUS       sequence-340180-        3204 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB5r/attB1r entry clone for generation of N-terminal 
            Nanoluciferase fusion proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3204)
  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 3204)
  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: C511-E02: attB5r-NanoLuc-nostop-attB1r
FEATURES             Location/Qualifiers
     source          1..3204
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(62..185)
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    708..832
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(904..923)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(905..923)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(928..944)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(928..944)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1369..2160
                     /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      2253..2841
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2742..2761
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2976..3003)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3095..3181)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gcgttaacgc taccatggag ctcgaattct acagtcacta ataccatcta agtagttgat
       61 tcatagtgac tgcatatgtt gtgttttaca gtattatgta gtctgttttt tatgcaaaat
      121 ctaatttaat atattgatat ttatatcatt ttacgtttct cgttcaactt ttgtatacaa
      181 agtggaccat ggtgtttact ctcgaggatt tcgtcggaga ttggcgccag accgccggtt
      241 acaaccttga ccaagtgctc gaacagggag gagtcagctc actgttccag aatctcgggg
      301 tgtctgtgac tcctatccag cgcatcgtcc tttctggcga aaacgggctt aagatcgaca
      361 tccatgtgat tattccctac gagggactgt caggcgacca gatggggcaa attgaaaaga
      421 ttttcaaggt ggtgtatccg gtcgacgatc accacttcaa ggtgatcctc cattatggga
      481 ccctggtgat tgacggagtg accccgaata tgatcgacta cttcggtcgg ccttacgaag
      541 gcattgctgt gttcgacgga aagaaaatca ctgtcaccgg taccctctgg aatggtaaca
      601 aaatcattga cgagagactg atcaaccctg acggatcact tctgttccgg gtcaccatca
      661 atggggtgac cggttggagg ctgtgcgagc gcatcctggc cggatccaca actttgtaca
      721 aaaaagttga acgagaaacg taaaatgata taaatatcaa tatattaaat tagattttgc
      781 ataaaaaaca gactacataa tactgtaaaa cacaacatat gcagtcacta tgaaccaact
      841 acttagatgg tattagtgac ctgtagaatt cgagctctag agctgcaggg cggccgcgat
      901 atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg
      961 tctcaaaatc tctgatgtta cattgcacaa gatcggtacc cggggatcct ctagaccagc
     1021 caggacagaa atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg cacaccgtgg
     1081 aaacggatga aggcacgaac ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc
     1141 aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac
     1201 ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttggggtaca gtctatgcct
     1261 cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc
     1321 aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taaacattat gagggaagcg
     1381 gtgatcgccg aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc
     1441 gaaccgacgt tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca
     1501 cacagtgata ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga
     1561 gctttgatca acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc
     1621 gctgtagaag tcaccattgt tgtgcacgac gacatcattc cgtggcgtta tccagctaag
     1681 cgcgaactgc aatttggaga atggcagcgc aatgacattc ttgcaggtat cttcgagcca
     1741 gccacgatcg acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc
     1801 ttggtaggtc cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag
     1861 gcgctaaatg aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga
     1921 aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg
     1981 aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata
     2041 cttgaagcta gacaggctta tcttggacaa gaagaagatc gcttggcctc gcgcgcagat
     2101 cagttggaag aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa
     2161 ccctcgagcc acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg
     2221 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2281 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     2341 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     2401 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2461 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2521 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2581 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     2641 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     2701 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     2761 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     2821 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     2881 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     2941 attaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct
     3001 tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc
     3061 gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca
     3121 ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt
     3181 gatgcctggc agttccctac tctc
//
