LOCUS       sequence-330604-        3598 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB4/attB1r 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 3598)
  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 3598)
  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: C413-E11: attB4-Grp78p>-attB1r
FEATURES             Location/Qualifiers
     source          1..3598
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    593..688
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     enhancer        691..994
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    1643..1767
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1839..1858)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1840..1858)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1863..1879)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1863..1879)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1981..2000)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1992..2801
                     /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(2059..2078)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2948..3536
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3437..3456
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccga gttaacgcta ccatggagct ccaaataatg
      601 attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgataagca atgctttttt
      661 ataatgccaa ctttgtatag aaaagttggc cgttacataa cttacggtaa atggcccgcc
      721 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      781 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      841 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      901 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      961 gtacatctac gtattagtca tcgctattac catgatgatg cggttttggc agtacatcaa
     1021 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     1081 tgggagtttg ttttgactag ttaccggcgg aaacggtctc gggttgagag gtcacccgag
     1141 ggacaggcag ctgctgaacc aataggaccg gcgcacaggg cggatgctgc ccctcattgg
     1201 cggccgttga gagtgaccaa gagccaatga gtcagcccgg ggggcgtagc agtgacgtaa
     1261 gttgcggagg aggccgcttc gaatcggcag cggccagctt ggtggcatgg accaatcagc
     1321 gtcctccaac gaggagcgcc ttcgccaatc ggaggcctcc acgacggggc tggggggagg
     1381 gtatataagc cgagtcggcg gcggcgcgct ccacacgggc cgagaccaca gcgacgggag
     1441 cgtctgcctc tgcggggccg agaggtaagc gccgcggcct gccctttcca ggccaactcg
     1501 gagcccgtct cgtggctccg cctgatcggg ggctcctgtc gccctcagat cggtcggaac
     1561 gccgtcgcgc tccgggacta caagcctgtt gctgggcccg gagactgccg aaggaccgct
     1621 gagcactgtc ctcagcgccg gcacaacttt gtacaaaaaa gttgaacgag aaacgtaaaa
     1681 tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta cataatactg
     1741 taaaacacaa catatgcagt cactatgaac caactactta gatggtatta gtgacctgta
     1801 gaattcgagc tctagagctg cagggcggcc gcgatatccc ctatagtgag tcgtattaca
     1861 tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg
     1921 cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata
     1981 caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca
     2041 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     2101 caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     2161 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
     2221 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     2281 ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     2341 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
     2401 gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg
     2461 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     2521 ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt
     2581 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     2641 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     2701 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     2761 ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt
     2821 tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat
     2881 cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     2941 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3001 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3061 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     3121 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3181 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3241 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3301 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     3361 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     3421 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     3481 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     3541 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtt
//
