LOCUS       sequence-331911-        2970 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB4/attB5 entry clone for mammalian expression using the 
            CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2970)
  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 2970)
  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: C453-E02: attB4-CAGp>-attB5
FEATURES             Location/Qualifiers
     source          1..2970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(60..146)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    194..289
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     enhancer        300..603
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    complement(1011..1106)
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1225..1244)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1236..2045
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1303..1322)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2188..2776
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2677..2696
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2911..2938)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga
       61 tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
      121 tccgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct
      181 accatggagc tccaaataat gattttattt tgactgatag tgacctgttc gttgcaacaa
      241 attgataagc aatgcttttt tataatgcca actttgtata gaaaagttgg ccctgcaggc
      301 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      361 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      421 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      481 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      541 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattaac
      601 atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc
      661 ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc ggggcggggc
      721 gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat
      781 ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc
      841 tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct
      901 ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg
      961 taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcggc caacttttgt
     1021 atacaaagtt ggcattataa aaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     1081 tatcagtcaa aataaaatca ttatttggag ctctagagct gcagggcgca gggcccgtgt
     1141 ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac
     1201 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt
     1261 cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
     1321 ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag
     1381 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
     1441 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
     1501 ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc caggtattag
     1561 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
     1621 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc
     1681 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
     1741 atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcgccgg
     1801 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
     1861 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
     1921 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     1981 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
     2041 tctaatcaga attggttaat tggttgtaac accggcagag cattacgctg acttgacggg
     2101 acggcgcaag ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2161 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2221 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2281 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     2341 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     2401 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     2461 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     2521 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     2581 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     2641 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     2701 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     2761 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     2821 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     2881 cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc
     2941 ttagtttgat gcctggcagt ttatggcggg
//
