LOCUS       sequence-330545-        3405 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB5r/attB1r entry clone for generation of N-terminal 
            mClover3 fusion proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3405)
  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 3405)
  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: C512-E22: attB5r-mClover3-nostop-attB1r
FEATURES             Location/Qualifiers
     source          1..3405
                     /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"
     protein_bind    891..1015
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1087..1106)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1088..1106)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1111..1127)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1111..1127)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1552..2343
                     /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      2436..3024
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2925..2944
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3159..3186)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3278..3364)
                     /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 aaagtgggac cgcatggtgt
      181 cgaagggcga agaactcttt accggagtgg tgccgattct ggtggaactg gacggagatg
      241 tcaacggtca taagttctcc gtccggggag aaggagaggg cgatgccact aacgggaagc
      301 tcacgctgaa attcatctgc accactggga agcttccggt gccctggcct accctcgtca
      361 ccaccttcgg ttacggggtg gcctgtttct cacggtaccc agaccacatg aaacaacatg
      421 acttctttaa gtccgcaatg cccgagggat acgtgcagga acgcaccatc agcttcaagg
      481 acgacggcac ttacaagact agagccgaag tcaagttcga gggagataca ctcgtgaaca
      541 ggatcgaact gaagggcatt gatttcaagg aggacggcaa cattctgggc cacaagttgg
      601 agtacaactt caattcccac tacgtgtaca tcaccgcgga caagcagaag aactgcatca
      661 aggccaactt caagatccgc cacaacgtgg aggatggatc ggtccagctg gcggatcact
      721 accagcaaaa cacccccatt ggcgacggtc ctgtgctgct gcctgacaac cactacctga
      781 gccaccagtc caagttgtcc aaagacccga atgagaagcg ggaccacatg gtgctgctgg
      841 aattcgtgac tgccgctgga atcacccatg ggatggacga gctgtataag acaactttgt
      901 acaaaaaagt tgaacgagaa acgtaaaatg atataaatat caatatatta aattagattt
      961 tgcataaaaa acagactaca taatactgta aaacacaaca tatgcagtca ctatgaacca
     1021 actacttaga tggtattagt gacctgtaga attcgagctc tagagctgca gggcggccgc
     1081 gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc
     1141 gtgtctcaaa atctctgatg ttacattgca caagatcggt acccggggat cctctagacc
     1201 agccaggaca gaaatgcctc gacttcgctg ctacccaagg ttgccgggtg acgcacaccg
     1261 tggaaacgga tgaaggcacg aacccagtgg acataagcct gttcggttcg taagctgtaa
     1321 tgcaagtagc gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt
     1381 aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttggggt acagtctatg
     1441 cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc
     1501 agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaaacat tatgagggaa
     1561 gcggtgatcg ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat
     1621 ctcgaaccga cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag
     1681 ccacacagtg atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg
     1741 cgagctttga tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc
     1801 cgcgctgtag aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct
     1861 aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag
     1921 ccagccacga tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt
     1981 gccttggtag gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt
     2041 gaggcgctaa atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag
     2101 cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg
     2161 ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc
     2221 atacttgaag ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca
     2281 gatcagttgg aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa
     2341 taaccctcga gccacccatg accaaaatcc cttaacgtga gttacgcgtc gttccactga
     2401 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     2461 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     2521 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     2581 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     2641 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     2701 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     2761 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     2821 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     2881 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     2941 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     3001 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     3061 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     3121 cgtattaccg ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg
     3181 ccttctgctt agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg
     3241 gccgttgctt cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt
     3301 tcaccgacaa acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta
     3361 tttgatgcct ggcagttccc tactctcgcg ttaacgctac catgg
//
