LOCUS       sequence-330547-        3381 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB5r/attB1r entry clone for generation of N-terminal 
            mKate2 fusion proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3381)
  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 3381)
  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-E29: attB5r-mKate2-nostop-attB1r
FEATURES             Location/Qualifiers
     source          1..3381
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(14..100)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    complement(185..308)
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    1008..1132
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1204..1223)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1205..1223)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1228..1244)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1228..1244)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1669..2460
                     /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      2553..3141
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3042..3061
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3276..3303)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
       61 gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact
      121 ctcgcgttaa cgctaccatg gagctcgaat tctacagtca ctaataccat ctaagtagtt
      181 gattcatagt gactgcatat gttgtgtttt acagtattat gtagtctgtt ttttatgcaa
      241 aatctaattt aatatattga tatttatatc attttacgtt tctcgttcaa cttttgtata
      301 caaagtggac catggtgtcg gaattgatta aggaaaatat gcatatgaag ctttacatgg
      361 agggaacagt gaacaaccat cacttcaagt gcacctccga gggcgaagga aagccttacg
      421 agggaaccca gactatgcgg attaaggctg tcgagggcgg cccgctgcct tttgcgttcg
      481 acattctggc cacctccttt atgtacggat ccaagacttt catcaaccac acgcagggaa
      541 tcccggattt cttcaagcaa agcttccccg agggtttcac ctgggagcgc gtcaccactt
      601 acgaggacgg gggcgtgctg accgcgactc aggataccag cctgcaagac ggctgcctca
      661 tctataacgt gaagatccgc ggagtgaact tcccgtcgaa cgggccagtg atgcagaaga
      721 aaaccctggg ttgggaagca tccaccgaaa ctctgtaccc cgctgacggt ggactcgagg
      781 gaagggccga tatggccctg aagcttgtgg gcgggggcca cctgatctgc aacctcaaga
      841 ccacctaccg gtcaaagaaa cccgccaaga acctgaagat gcctggggtc tactacgtgg
      901 accgccggct ggagagaatc aaggaagccg acaaggaaac ttacgtggaa cagcacgaag
      961 tggccgtggc cagatactgt gacctcccgt ccaaactggg acaccggaca actttgtaca
     1021 aaaaagttga acgagaaacg taaaatgata taaatatcaa tatattaaat tagattttgc
     1081 ataaaaaaca gactacataa tactgtaaaa cacaacatat gcagtcacta tgaaccaact
     1141 acttagatgg tattagtgac ctgtagaatt cgagctctag agctgcaggg cggccgcgat
     1201 atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg
     1261 tctcaaaatc tctgatgtta cattgcacaa gatcggtacc cggggatcct ctagaccagc
     1321 caggacagaa atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg cacaccgtgg
     1381 aaacggatga aggcacgaac ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc
     1441 aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac
     1501 ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttggggtaca gtctatgcct
     1561 cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc
     1621 aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taaacattat gagggaagcg
     1681 gtgatcgccg aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc
     1741 gaaccgacgt tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca
     1801 cacagtgata ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga
     1861 gctttgatca acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc
     1921 gctgtagaag tcaccattgt tgtgcacgac gacatcattc cgtggcgtta tccagctaag
     1981 cgcgaactgc aatttggaga atggcagcgc aatgacattc ttgcaggtat cttcgagcca
     2041 gccacgatcg acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc
     2101 ttggtaggtc cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag
     2161 gcgctaaatg aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga
     2221 aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg
     2281 aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata
     2341 cttgaagcta gacaggctta tcttggacaa gaagaagatc gcttggcctc gcgcgcagat
     2401 cagttggaag aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa
     2461 ccctcgagcc acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg
     2521 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2581 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     2641 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     2701 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2761 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2821 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2881 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     2941 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3001 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3061 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3121 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     3181 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     3241 attaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct
     3301 tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc
     3361 gttgcttcac aacgttcaaa t
//
