LOCUS       sequence-333339-        4335 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 4335)
  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 4335)
  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-E15: attB5r-ffluc2-nostop-attB1r
FEATURES             Location/Qualifiers
     source          1..4335
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1609..1733
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1805..1824)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1806..1824)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1829..1845)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1829..1845)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2270..3061
                     /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      3154..3742
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3643..3662
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3877..3904)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3996..4082)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    complement(4167..4290)
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(4294..4335,1..1608)
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
ORIGIN
        1 tacccactcg aagacgggac cgccggcgag cagctgcaca aagccatgaa gcgctacgcc
       61 ctggtgcccg gcaccatcgc ctttaccgac gcacatatcg aggtggacat tacctacgcc
      121 gagtacttcg agatgagcgt tcggctggca gaagctatga agcgctatgg gctgaataca
      181 aaccatcgga tcgtggtgtg cagcgagaat agcttgcagt tcttcatgcc cgtgttgggt
      241 gccctgttca tcggtgtggc tgtggcccca gctaacgaca tctacaacga gcgcgagctg
      301 ctgaacagca tgggcatcag ccagcccacc gtcgtattcg tgagcaagaa agggctgcaa
      361 aagatcctca acgtgcaaaa gaagctaccg atcatacaaa agatcatcat catggatagc
      421 aagaccgact accagggctt ccaaagcatg tacaccttcg tgacttccca tttgccaccc
      481 ggcttcaacg agtacgactt cgtgcccgag agcttcgacc gggacaaaac catcgccctg
      541 atcatgaaca gtagtggcag taccggattg cccaagggcg tagccctacc gcaccgcacc
      601 gcttgtgtcc gattcagtca tgcccgcgac cccatcttcg gcaaccagat catccccgac
      661 accgctatcc tcagcgtggt gccatttcac cacggcttcg gcatgttcac cacgctgggc
      721 tacttgatct gcggctttcg ggtcgtgctc atgtaccgct tcgaggagga gctattcttg
      781 cgcagcttgc aagactataa gattcaatct gccctgctgg tgcccacact atttagcttc
      841 ttcgctaaga gcactctcat cgacaagtac gacctaagca acttgcacga gatcgccagc
      901 ggcggggcgc cgctcagcaa ggaggtaggt gaggccgtgg ccaaacgctt ccacctacca
      961 ggcatccgcc agggctacgg cctgacagaa acaaccagcg ccattctgat cacccccgaa
     1021 ggggacgaca agcctggcgc agtaggcaag gtggtgccct tcttcgaggc taaggtggtg
     1081 gacttggaca ccggtaagac actgggtgtg aaccagcgcg gcgagctgtg cgtccgtggc
     1141 cccatgatca tgagcggcta cgttaacaac cccgaggcta caaacgctct catcgacaag
     1201 gacggctggc tgcacagcgg cgacatcgcc tactgggacg aggacgagca cttcttcatc
     1261 gtggaccggc tgaagagcct gatcaaatac aagggctacc aggtagcccc agccgaactg
     1321 gagagcatcc tgctgcaaca ccccaacatc ttcgacgccg gggtcgccgg cctgcccgac
     1381 gacgatgccg gcgagctgcc cgccgcagtc gtcgtgctgg aacacggtaa aaccatgacc
     1441 gagaaggaga tcgtggacta tgtggccagc caggttacaa ccgccaagaa gctgcgcggt
     1501 ggtgttgtgt tcgtggacga ggtgcctaaa ggactgaccg gcaagttgga cgcccgcaag
     1561 atccgcgaga ttctcattaa ggccaagaag ggcggcaaga tcgccgtgac aactttgtac
     1621 aaaaaagttg aacgagaaac gtaaaatgat ataaatatca atatattaaa ttagattttg
     1681 cataaaaaac agactacata atactgtaaa acacaacata tgcagtcact atgaaccaac
     1741 tacttagatg gtattagtga cctgtagaat tcgagctcta gagctgcagg gcggccgcga
     1801 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     1861 gtctcaaaat ctctgatgtt acattgcaca agatcggtac ccggggatcc tctagaccag
     1921 ccaggacaga aatgcctcga cttcgctgct acccaaggtt gccgggtgac gcacaccgtg
     1981 gaaacggatg aaggcacgaa cccagtggac ataagcctgt tcggttcgta agctgtaatg
     2041 caagtagcgt atgcgctcac gcaactggtc cagaaccttg accgaacgca gcggtggtaa
     2101 cggcgcagtg gcggttttca tggcttgtta tgactgtttt tttggggtac agtctatgcc
     2161 tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag
     2221 caacgatgtt acgcagcagg gcagtcgccc taaaacaaag ttaaacatta tgagggaagc
     2281 ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg agcgccatct
     2341 cgaaccgacg ttgctggccg tacatttgta cggctccgca gtggatggcg gcctgaagcc
     2401 acacagtgat attgatttgc tggttacggt gaccgtaagg cttgatgaaa caacgcggcg
     2461 agctttgatc aacgaccttt tggaaacttc ggcttcccct ggagagagcg agattctccg
     2521 cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa
     2581 gcgcgaactg caatttggag aatggcagcg caatgacatt cttgcaggta tcttcgagcc
     2641 agccacgatc gacattgatc tggctatctt gctgacaaaa gcaagagaac atagcgttgc
     2701 cttggtaggt ccagcggcgg aggaactctt tgatccggtt cctgaacagg atctatttga
     2761 ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc gactgggctg gcgatgagcg
     2821 aaatgtagtg cttacgttgt cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc
     2881 gaaggatgtc gctgccgact gggcaatgga gcgcctgccg gcccagtatc agcccgtcat
     2941 acttgaagct agacaggctt atcttggaca agaagaagat cgcttggcct cgcgcgcaga
     3001 tcagttggaa gaatttgtcc actacgtgaa aggcgagatc accaaggtag tcggcaaata
     3061 accctcgagc cacccatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     3121 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3181 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3241 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3301 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3361 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3421 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3481 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3541 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3601 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3661 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3721 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3781 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     3841 tattaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc
     3901 ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc
     3961 cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc
     4021 accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt
     4081 tgatgcctgg cagttcccta ctctcgcgtt aacgctacca tggagctcga attctacagt
     4141 cactaatacc atctaagtag ttgattcata gtgactgcat atgttgtgtt ttacagtatt
     4201 atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg
     4261 tttctcgttc aacttttgta tacaaagtgg accatggaag atgccaaaaa cattaagaag
     4321 ggcccagcgc cattc
//
