LOCUS       sequence-340176-        2673 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB4/attB5 entry clone for mammalian expression using the 
            human EF1alpha promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2673)
  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 2673)
  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-E21: attB4-FerHp>-attB5
FEATURES             Location/Qualifiers
     source          1..2673
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    50..145
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     enhancer        333..524
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     481..501
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     protein_bind    complement(570..665)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      713..799
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      891..918
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      complement(1053..1641)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1133..1152)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1784..2593)
                     /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     2507..2526
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2585..2604
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 atcagagatt ttgagacacg ggccctgcgc cctgcagctc tagagctcca aataatgatt
       61 ttattttgac tgatagtgac ctgttcgttg caacaaattg ataagcaatg cttttttata
      121 atgccaactt tgtatacaaa agttggccgg cgaagaacgt ctggccctgc gggtcgcttg
      181 tggtctctta tagccgcgtc ggcgtcaggc ccgccccggc caatcagcgc cgcccgcccc
      241 gagcccgctc cttccggtgg gcgcgggacc ccgcccctgc tgtgggggag gggcggccgc
      301 tggaggccct cgcgcgctct ggcggaacta gtgggactat ggttgctgac taattgagat
      361 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg
      421 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
      481 accctaactg acacacattc cacagctggt tctttcagcc tcagaaggta cctaaccaag
      541 ttcctctttc agaggttatt tcaggccgcc aacttttcta tacaaagttg gcattataaa
      601 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
      661 tatttggagc tccatggtag cgttaacgcg agagtaggga actgccaggc atcaaataaa
      721 acgaaaggct cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
      781 tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg
      841 agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat
      901 cctgacggat ggcctttttg cgtttctaca aactcttcct ggctagcggt aatacggtta
      961 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1021 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1081 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1141 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1201 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     1261 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1321 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1381 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1441 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     1501 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     1561 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1621 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     1681 tggaacgaaa actcacgtta agggattttg gtcatgagct tgcgccgtcc cgtcaagtca
     1741 gcgtaatgct ctgccggtgt tacaaccaat taaccaattc tgattagaaa aactcatcga
     1801 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     1861 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     1921 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     1981 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggcgagaatg
     2041 gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     2101 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
     2161 atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     2221 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     2281 atgctgtttt tccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     2341 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
     2401 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     2461 gcttcccata caagcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     2521 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctcgac gtttcccgtt
     2581 gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc
     2641 atgatgatat atttttatct tgtgcaatgt aac
//
