LOCUS       sequence-330523-        3391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB2r/attB3 entry clone for construction of C-terminal SNAP
            tagged fusion proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3391)
  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 3391)
  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: C234-E25: attB2r-SNAP-attB3
FEATURES             Location/Qualifiers
     source          1..3391
                     /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(852..947)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1358..2149
                     /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      2242..2830
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2731..2750
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2984..3001
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3160..3187)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3279..3365)
                     /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 agtgggcgac aaggactgcg agatgaaacg
      301 gactaccctt gattcgcccc tcggaaagct cgaactcagc ggctgtgaac agggtctgca
      361 cgagatcaag ctgctgggaa agggcacctc cgccgccgac gccgtggaag tgccagctcc
      421 cgccgccgtg cttggaggcc ccgagccgct gatgcaggcc actgcctggc tcaacgcgta
      481 ctttcaccaa ccggaagcca tcgaggaatt ccctgtgccc gcactgcacc acccggtgtt
      541 ccagcaggag tccttcaccc gccaagtcct gtggaagctg ttgaaggtcg tgaagttcgg
      601 cgaagtgatc agctaccagc agctggcagc cctggccggg aaccctgccg cgaccgctgc
      661 tgtcaagacc gcgctgtccg gcaacccagt gccgattctg attccctgcc atcgggtggt
      721 gtcatcctcg ggagccgtgg gaggctacga aggaggtttg gcggtcaagg agtggctgct
      781 ggcacatgag gggcacagac tcggaaagcc gggactgggg cctgccggga tcggtgcccc
      841 tggctcctaa tcaactttat tatacaaagt tggcattata aaaaagcatt gcttatcaat
      901 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgga gctccatggt
      961 agcgttaacg cggccgcgat atcggtaccc ggggatcctc tagaccagcc aggacagaaa
     1021 tgcctcgact tcgctgctac ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa
     1081 ggcacgaacc cagtggacat aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat
     1141 gcgctcacgc aactggtcca gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc
     1201 ggttttcatg gcttgttatg actgtttttt tggggtacag tctatgcctc gggcatccaa
     1261 gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgttac
     1321 gcagcagggc agtcgcccta aaacaaagtt aaacattatg agggaagcgg tgatcgccga
     1381 agtatcgact caactatcag aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt
     1441 gctggccgta catttgtacg gctccgcagt ggatggcggc ctgaagccac acagtgatat
     1501 tgatttgctg gttacggtga ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa
     1561 cgaccttttg gaaacttcgg cttcccctgg agagagcgag attctccgcg ctgtagaagt
     1621 caccattgtt gtgcacgacg acatcattcc gtggcgttat ccagctaagc gcgaactgca
     1681 atttggagaa tggcagcgca atgacattct tgcaggtatc ttcgagccag ccacgatcga
     1741 cattgatctg gctatcttgc tgacaaaagc aagagaacat agcgttgcct tggtaggtcc
     1801 agcggcggag gaactctttg atccggttcc tgaacaggat ctatttgagg cgctaaatga
     1861 aaccttaacg ctatggaact cgccgcccga ctgggctggc gatgagcgaa atgtagtgct
     1921 tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc
     1981 tgccgactgg gcaatggagc gcctgccggc ccagtatcag cccgtcatac ttgaagctag
     2041 acaggcttat cttggacaag aagaagatcg cttggcctcg cgcgcagatc agttggaaga
     2101 atttgtccac tacgtgaaag gcgagatcac caaggtagtc ggcaaataac cctcgagcca
     2161 cccatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt
     2221 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2281 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2341 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     2401 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2461 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     2521 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     2581 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     2641 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     2701 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     2761 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     2821 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     2881 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     2941 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     3001 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     3061 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     3121 tacgcgtacc gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg
     3181 gccttctgct tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg
     3241 ggccgttgct tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg
     3301 ttcaccgaca aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt
     3361 atttgatgcc tggcagttcc ctactctcgc g
//
