LOCUS       sequence-331910-        2834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB4/attB5 entry clone for mammalian expression using an 
            optimized human CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2834)
  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 2834)
  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-E04: attB4-CMV51p>-attB5
FEATURES             Location/Qualifiers
     source          1..2834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(59..145)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    193..288
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     enhancer        299..602
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        603..806
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     757..777
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     803..827
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    complement(874..969)
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1088..1107)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1099..1908
                     /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(1166..1185)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2051..2639
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2540..2559
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2774..2801)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat
       61 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
      121 ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta
      181 ccatggagct ccaaataatg attttatttt gactgatagt gacctgttcg ttgcaacaaa
      241 ttgataagca atgctttttt ataatgccaa ctttgtatag aaaagttggc cctgcaggcg
      301 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      361 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      421 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      481 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      541 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      601 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      661 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      721 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      781 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc
      841 atccacgctg ttttgacctc catagaagac agccaacttt tgtatacaaa gttggcatta
      901 taaaaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa
      961 tcattatttg gagctctaga gctgcagggc gcagggcccg tgtctcaaaa tctctgatgt
     1021 tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac
     1081 agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc gcgattaaat
     1141 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
     1201 ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
     1261 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
     1321 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
     1381 ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata tcctgattca
     1441 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt
     1501 tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg
     1561 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
     1621 caagtctgga aagaaatgca taaacttttg ccattctcgc cggattcagt cgtcactcat
     1681 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat
     1741 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc
     1801 ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct
     1861 gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agaattggtt
     1921 aattggttgt aacaccggca gagcattacg ctgacttgac gggacggcgc aagctcatga
     1981 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     2041 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     2101 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     2161 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     2221 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     2281 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     2341 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     2401 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     2461 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     2521 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     2581 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     2641 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     2701 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgctagc caggaagagt
     2761 ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt gatgcctggc
     2821 agtttatggc gggc
//
