LOCUS       sequence-330546-        3399 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB5r/attB1r entry clone for generation of N-terminal 
            mRuby3 fusion proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3399)
  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 3399)
  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-E23: attB5r-mRuby3-nostop-attB1r
FEATURES             Location/Qualifiers
     source          1..3399
                     /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    885..1009
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1081..1100)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1082..1100)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1105..1121)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1105..1121)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1546..2337
                     /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      2430..3018
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2919..2938
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3153..3180)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3272..3358)
                     /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 cgaaggggga ggaactcatt aaggaaaaca tgcgcatgaa ggtcgtgatg gagggttccg
      241 tgaatggcca ccagtttaag tgcaccggag aaggggaagg acgcccgtac gaaggggtgc
      301 aaacaatgcg gatcaaggtc atcgaaggcg gacctctgcc tttcgcgttc gacatcctcg
      361 ccacttcctt tatgtacggg agccggactt tcatcaagta cccggccgac attcccgatt
      421 tcttcaagca atcattcccg gaaggcttca cctgggagag agtgaccaga tacgaggatg
      481 ggggagtggt caccgtgacg caggacactt cgctggagga cggagagctg gtgtacaacg
      541 tgaaggtccg cggagtgaac ttcccctcca acggaccagt gatgcagaaa aagactaagg
      601 gctgggaacc caacaccgaa atgatgtacc ctgccgatgg cggtttgagg ggatacaccg
      661 acatcgcact gaaggtcgac ggcggtggac acctccactg caacttcgtc accacctacc
      721 ggtccaagaa aaccgtgggc aacatcaaga tgccgggagt gcatgccgtg gaccacagac
      781 ttgagcgcat tgaggagagc gacaacgaaa cctacgtggt gcagcgggaa gtggctgtgg
      841 ccaagtacag caacctgggt ggcggcatgg atgagctgta taagacaact ttgtacaaaa
      901 aagttgaacg agaaacgtaa aatgatataa atatcaatat attaaattag attttgcata
      961 aaaaacagac tacataatac tgtaaaacac aacatatgca gtcactatga accaactact
     1021 tagatggtat tagtgacctg tagaattcga gctctagagc tgcagggcgg ccgcgatatc
     1081 ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct
     1141 caaaatctct gatgttacat tgcacaagat cggtacccgg ggatcctcta gaccagccag
     1201 gacagaaatg cctcgacttc gctgctaccc aaggttgccg ggtgacgcac accgtggaaa
     1261 cggatgaagg cacgaaccca gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag
     1321 tagcgtatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg tggtaacggc
     1381 gcagtggcgg ttttcatggc ttgttatgac tgtttttttg gggtacagtc tatgcctcgg
     1441 gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac
     1501 gatgttacgc agcagggcag tcgccctaaa acaaagttaa acattatgag ggaagcggtg
     1561 atcgccgaag tatcgactca actatcagag gtagttggcg tcatcgagcg ccatctcgaa
     1621 ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg atggcggcct gaagccacac
     1681 agtgatattg atttgctggt tacggtgacc gtaaggcttg atgaaacaac gcggcgagct
     1741 ttgatcaacg accttttgga aacttcggct tcccctggag agagcgagat tctccgcgct
     1801 gtagaagtca ccattgttgt gcacgacgac atcattccgt ggcgttatcc agctaagcgc
     1861 gaactgcaat ttggagaatg gcagcgcaat gacattcttg caggtatctt cgagccagcc
     1921 acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag cgttgccttg
     1981 gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct atttgaggcg
     2041 ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga tgagcgaaat
     2101 gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat cgcgccgaag
     2161 gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc cgtcatactt
     2221 gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg cgcagatcag
     2281 ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg caaataaccc
     2341 tcgagccacc catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca
     2401 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2461 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2521 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     2581 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2641 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2701 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2761 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2821 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2881 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     2941 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3001 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3061 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3121 accgctagcc aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct
     3181 gcttagtttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt
     3241 gcttcacaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg
     3301 acaaacaaca gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat
     3361 gcctggcagt tccctactct cgcgttaacg ctaccatgg
//
