LOCUS       sequence-330925-        3532 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB2r/attB3 entry clone for construction of C-terminal 
            mRuby3 fusion proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3532)
  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 3532)
  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: C232-E30: attB2r-mRuby3-attB3
FEATURES             Location/Qualifiers
     source          1..3532
                     /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"
     protein_bind    complement(993..1088)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1499..2290
                     /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      2383..2971
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2872..2891
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3125..3142
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3301..3328)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3420..3506)
                     /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 agtgggcggg gtgtcgaagg gggaggaact
      301 cattaaggaa aacatgcgca tgaaggtcgt gatggagggt tccgtgaatg gccaccagtt
      361 taagtgcacc ggagaagggg aaggacgccc gtacgaaggg gtgcaaacaa tgcggatcaa
      421 ggtcatcgaa ggcggacctc tgcctttcgc gttcgacatc ctcgccactt cctttatgta
      481 cgggagccgg actttcatca agtacccggc cgacattccc gatttcttca agcaatcatt
      541 cccggaaggc ttcacctggg agagagtgac cagatacgag gatgggggag tggtcaccgt
      601 gacgcaggac acttcgctgg aggacggaga gctggtgtac aacgtgaagg tccgcggagt
      661 gaacttcccc tccaacggac cagtgatgca gaaaaagact aagggctggg aacccaacac
      721 cgaaatgatg taccctgccg atggcggttt gaggggatac accgacatcg cactgaaggt
      781 cgacggcggt ggacacctcc actgcaactt cgtcaccacc taccggtcca agaaaaccgt
      841 gggcaacatc aagatgccgg gagtgcatgc cgtggaccac agacttgagc gcattgagga
      901 gagcgacaac gaaacctacg tggtgcagcg ggaagtggct gtggccaagt acagcaacct
      961 gggtggcggc atggatgagc tgtataagta atcaacttta ttatacaaag ttggcattat
     1021 aaaaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat
     1081 cattatttgg agctccatgg tagcgttaac gcggccgcga tatcggtacc cggggatcct
     1141 ctagaccagc caggacagaa atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg
     1201 cacaccgtgg aaacggatga aggcacgaac ccagtggaca taagcctgtt cggttcgtaa
     1261 gctgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag
     1321 cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttggggtaca
     1381 gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt
     1441 atggagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taaacattat
     1501 gagggaagcg gtgatcgccg aagtatcgac tcaactatca gaggtagttg gcgtcatcga
     1561 gcgccatctc gaaccgacgt tgctggccgt acatttgtac ggctccgcag tggatggcgg
     1621 cctgaagcca cacagtgata ttgatttgct ggttacggtg accgtaaggc ttgatgaaac
     1681 aacgcggcga gctttgatca acgacctttt ggaaacttcg gcttcccctg gagagagcga
     1741 gattctccgc gctgtagaag tcaccattgt tgtgcacgac gacatcattc cgtggcgtta
     1801 tccagctaag cgcgaactgc aatttggaga atggcagcgc aatgacattc ttgcaggtat
     1861 cttcgagcca gccacgatcg acattgatct ggctatcttg ctgacaaaag caagagaaca
     1921 tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt gatccggttc ctgaacagga
     1981 tctatttgag gcgctaaatg aaaccttaac gctatggaac tcgccgcccg actgggctgg
     2041 cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa
     2101 aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg cccagtatca
     2161 gcccgtcata cttgaagcta gacaggctta tcttggacaa gaagaagatc gcttggcctc
     2221 gcgcgcagat cagttggaag aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt
     2281 cggcaaataa ccctcgagcc acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     2341 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2401 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     2461 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2521 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2581 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2641 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     2701 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     2761 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     2821 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     2881 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     2941 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3001 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     3061 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     3121 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     3181 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     3241 cagtgagcgc aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca
     3301 aaaaggccat ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg
     3361 cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga
     3421 tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
     3481 tccgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg cg
//
