LOCUS       sequence-330900-        3216 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB4/attB5 entry clone for mammalian expression using the 
            human Grp78 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3216)
  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 3216)
  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-E09: attB4-Grp78p>-attB5
FEATURES             Location/Qualifiers
     source          1..3216
                     /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"
     enhancer        756..1059
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    complement(1070..1165)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      1213..1299
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1391..1418
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      complement(1553..2141)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1633..1652)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2284..3093)
                     /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     3007..3026
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3085..3104
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 agctccaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgata
       61 agcaatgctt ttttataatg ccaactttgt atacaaaagt tggcggtgcc ggcgctgagg
      121 acagtgctca gcggtccttc ggcagtctcc gggcccagca acaggcttgt agtcccggag
      181 cgcgacggcg ttccgaccga tctgagggcg acaggagccc ccgatcaggc ggagccacga
      241 gacgggctcc gagttggcct ggaaagggca ggccgcggcg cttacctctc ggccccgcag
      301 aggcagacgc tcccgtcgct gtggtctcgg cccgtgtgga gcgcgccgcc gccgactcgg
      361 cttatatacc ctccccccag ccccgtcgtg gaggcctccg attggcgaag gcgctcctcg
      421 ttggaggacg ctgattggtc catgccacca agctggccgc tgccgattcg aagcggcctc
      481 ctccgcaact tacgtcactg ctacgccccc cgggctgact cattggctct tggtcactct
      541 caacggccgc caatgagggg cagcatccgc cctgtgcgcc ggtcctattg gttcagcagc
      601 tgcctgtccc tcgggtgacc tctcaacccg agaccgtttc cgccggtaac tagtcaaaac
      661 aaactcccat tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc aaaccgctat
      721 ccacgcccat tgatgtactg ccaaaaccgc atcatcatgg taatagcgat gactaatacg
      781 tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg
      841 ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt
      901 actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct
      961 attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc
     1021 ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ctgcagggcc aacttttcta
     1081 tacaaagttg gcattataaa aaagcattgc ttatcaattt gttgcaacga acaggtcact
     1141 atcagtcaaa ataaaatcat tatttggagc tccatggtag cgttaacgcg agagtaggga
     1201 actgccaggc atcaaataaa acgaaaggct cagtcggaag actgggcctt tcgttttatc
     1261 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac
     1321 gttgtgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat
     1381 caaactaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttcct
     1441 ggctagcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     1501 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     1561 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     1621 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     1681 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     1741 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     1801 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     1861 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     1921 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     1981 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2041 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2101 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2161 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagct
     2221 tgcgccgtcc cgtcaagtca gcgtaatgct ctgccggtgt tacaaccaat taaccaattc
     2281 tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
     2341 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
     2401 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
     2461 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
     2521 gactgaatcc ggcgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg
     2581 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
     2641 ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
     2701 cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg
     2761 atattcttct aatacctgga atgctgtttt tccggggatc gcagtggtga gtaaccatgc
     2821 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
     2881 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
     2941 aaacaactct ggcgcatcgg gcttcccata caagcgatag attgtcgcac ctgattgccc
     3001 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
     3061 cggcctcgac gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta
     3121 agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt aacatcagag
     3181 attttgagac acgggccctg cgccctgcag ctctag
//
