LOCUS       sequence-333336-        3020 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB4/attB1r entry clone for mammalian expression using the 
            minimal CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3020)
  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 3020)
  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: C413-E34: attB4-mCMVp>-attB1r
FEATURES             Location/Qualifiers
     source          1..3020
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(34..61)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(153..239)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     288..310
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     302..319
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     303..319
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    375..470
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     506..530
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    591..715
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(787..806)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(788..806)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(811..827)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(811..827)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1252..2043
                     /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      2136..2724
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2625..2644
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2878..2895
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc
       61 tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt
      121 tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc
      181 gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga
      241 tgcctggcag ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg
      301 ttgtaaaacg acggccagtc ttaaaattcg agctcggtcc gatcgggccc gcgttaacgc
      361 taccatggag ctccaaataa tgattttatt ttgactgata gtgacctgtt cgttgcaaca
      421 aattgataag caatgctttt ttataatgcc aactttgtat agaaaagttg gctaggcgtg
      481 tacggtggga ggcctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagac
      541 gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc acaactttgt
      601 acaaaaaagt tgaacgagaa acgtaaaatg atataaatat caatatatta aattagattt
      661 tgcataaaaa acagactaca taatactgta aaacacaaca tatgcagtca ctatgaacca
      721 actacttaga tggtattagt gacctgtaga attcgagctc tagagctgca gggcggccgc
      781 gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc
      841 gtgtctcaaa atctctgatg ttacattgca caagatcggt acccggggat cctctagacc
      901 agccaggaca gaaatgcctc gacttcgctg ctacccaagg ttgccgggtg acgcacaccg
      961 tggaaacgga tgaaggcacg aacccagtgg acataagcct gttcggttcg taagctgtaa
     1021 tgcaagtagc gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt
     1081 aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttggggt acagtctatg
     1141 cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc
     1201 agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaaacat tatgagggaa
     1261 gcggtgatcg ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat
     1321 ctcgaaccga cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag
     1381 ccacacagtg atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg
     1441 cgagctttga tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc
     1501 cgcgctgtag aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct
     1561 aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag
     1621 ccagccacga tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt
     1681 gccttggtag gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt
     1741 gaggcgctaa atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag
     1801 cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg
     1861 ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc
     1921 atacttgaag ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca
     1981 gatcagttgg aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa
     2041 taaccctcga gccacccatg accaaaatcc cttaacgtga gttacgcgtc gttccactga
     2101 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     2161 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     2221 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     2281 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     2341 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     2401 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     2461 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     2521 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     2581 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     2641 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     2701 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     2761 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     2821 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     2881 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     2941 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     3001 cgcaacgcaa ttaatacgcg
//
