LOCUS       sequence-330920-        3463 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB4/attB5 entry clone for mammalian expression using the 
            human Ubiquitin C promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3463)
  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 3463)
  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-E24: attB4-UbCp>-attB5
FEATURES             Location/Qualifiers
     source          1..3463
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    7..102
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(106..1317)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(250..269)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    complement(1317..1412)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      1460..1546
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1638..1665
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      complement(1800..2388)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1880..1899)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2531..3340)
                     /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     3254..3273
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3332..3351
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 agctccaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgata
       61 agcaatgctt ttttataatg ccaactttgt atacaaaagt tggccgtcta acaaaaaagc
      121 caaaaacggc cagaatttag cggacaattt actagtctaa cactgaaaat tacatattga
      181 cccaaatgat tacatttcaa aaggtgccta aaaaacttca caaaacacac tcgccaaccc
      241 cgagcgcata gttcaaaacc ggagcttcag ctacttaaga agataggtac ataaaaccga
      301 ccaaagaaac tgacgcctca cttatccctc ccctcaccag aggtccggcg cctgtcgatt
      361 caggagagcc taccctaggc ccgaaccctg cgtcctgcga cggagaaaag cctaccgcac
      421 acctaccggc aggtggcccc accctgcatt ataagccaac agaacgggtg acgtcacgac
      481 acgacgaggg cgcgcgctcc caaaggtacg ggtgcactgc ccaacggcac cgccataact
      541 gccgcccccg caacagacga caaaccgagt tctccagtca gtgacaaact tcacgtcagg
      601 gtccccagat ggtgccccag cccatctcac ccgaataaga gctttcccgc attagcgaag
      661 gcctcaagac cttgggttct tgccgcccac catgcccccc accttgtttc aacgacctca
      721 cagcccgcct cacaagcgtc ttccattcaa gactcgggaa cagccgccat tttgctgcgc
      781 tccccccaac ccccagttca gggcaacctt gctcgcggac ccagactaca gcccttggcg
      841 gtctctccac acgcttccgt cccaccgagc ggcccggcgg ccacgaaagc cccggccagc
      901 ccagcagccc gctactcacc aagtgacgat cacagcgatc cacaaacaag aaccgcgacc
      961 caaatcccgg ctgcgacgga actagctgtg ccacacccgg cgcgtcctta tataatcatc
     1021 ggcgttcacc gccccacgga gatccctccg cagaatcgcc gagaagggac tacttttcct
     1081 cgcctgttcc gctctctgga aagaaaacca gtgccctaga gtcacccaag tcccgtccta
     1141 aaatgtcctt ctgctgatac tggggttcta aggccgagtc ttatgagcag cgggccgctg
     1201 tcctgagcgt ccgggcggaa ggatcaggac gctcgctgcg cccttcgtct gacgtggcag
     1261 cgctcgccgt gaggaggggg gcgcccgcgg gaggcgccaa aacccggcgc ggaggccaac
     1321 ttttctatac aaagttggca ttataaaaaa gcattgctta tcaatttgtt gcaacgaaca
     1381 ggtcactatc agtcaaaata aaatcattat ttggagctcc atggtagcgt taacgcgaga
     1441 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcggaagact gggcctttcg
     1501 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga
     1561 tttgaacgtt gtgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc
     1621 caggcatcaa actaagcaga aggccatcct gacggatggc ctttttgcgt ttctacaaac
     1681 tcttcctggc tagcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     1741 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     1801 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     1861 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     1921 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     1981 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2041 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2101 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2161 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2221 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2281 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2341 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2401 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     2461 atgagcttgc gccgtcccgt caagtcagcg taatgctctg ccggtgttac aaccaattaa
     2521 ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag
     2581 gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga
     2641 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat
     2701 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat
     2761 gagtgacgac tgaatccggc gagaatggca aaagtttatg catttctttc cagacttgtt
     2821 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca
     2881 ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa
     2941 caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg
     3001 aatcaggata ttcttctaat acctggaatg ctgtttttcc ggggatcgca gtggtgagta
     3061 accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg
     3121 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat
     3181 gtttcagaaa caactctggc gcatcgggct tcccatacaa gcgatagatt gtcgcacctg
     3241 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat
     3301 ttaatcgcgg cctcgacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     3361 ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac
     3421 atcagagatt ttgagacacg ggccctgcgc cctgcagctc tag
//
