LOCUS       sequence-330603-        4066 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 4066)
  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 4066)
  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-E10: attB4-FerHp>-attB1r
FEATURES             Location/Qualifiers
     source          1..4066
                     /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        734..925
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(757..777)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     intron          1132..2079
                     /label=mEF-1-alpha intron
                     /note="intron upstream of the start codon of mouse 
                     EF-1-alpha"
     protein_bind    2111..2235
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2307..2326)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2308..2326)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2331..2347)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2331..2347)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2449..2468)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2460..3269
                     /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(2527..2546)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3416..4004
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3905..3924
                     /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 ggcctgaaat aacctctgaa agaggaactt
      721 ggttaggtac cttctgaggc tgaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
      781 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
      841 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
      901 tcaattagtc agcaaccata gtcccactag ttccgccaga gcgcgcgagg gcctccagcg
      961 gccgcccctc ccccacagca ggggcggggt cccgcgccca ccggaaggag cgggctcggg
     1021 gcgggcggcg ctgattggcc ggggcgggcc tgacgccgac gcggctataa gagaccacaa
     1081 gcgacccgca gggccagacg ttcttcgccg aagcttgccg tcagaacgca ggtgaggggc
     1141 gggtgtggct tccgcgggcc gccgagctgg aggtcctgct ccgagcgggc cgggccccgc
     1201 tgtcgtcggc ggggattagc tgcgagcatt cccgcttcga gttgcgggcg gcgcgggagg
     1261 cagagtgcga ggcctagcgg caaccccgta gcctcgcctc gtgtccggct tgaggcctag
     1321 cgtggtgtcc gcgccgccgc cgcgtgctac tccggccgca ctctggtctt tttttttttt
     1381 gttgttgttg ccctgctgcc ttcgattgcc gttcagcaat aggggctaac aaagggaggg
     1441 tgcggggctt gctcgcccgg agcccggaga ggtcatggtt ggggaggaat ggagggacag
     1501 gagtggcggc tggggcccgc ccgccttcgg agcacatgtc cgacgccacc tggatggggc
     1561 gaggcctggg gtttttcccg aagcaaccag gctggggtta gcgtgccgag gccatgtggc
     1621 cccagcaccc ggcacgatct ggcttggcgg cgccgcgttg ccctgcctcc ctaactaggg
     1681 tgaggccatc ccgtccggca ccagttgcgt gcgtggaaag atggccgctc ccgggccctg
     1741 ttgcaaggag ctcaaaatgg aggacgcggc agcccggtgg agcgggcggg tgagtcaccc
     1801 acacaaagga agagggcctg gtccctcacc ggctgctgct tcctgtgacc ccgtggtcct
     1861 atcggccgca atagtcacct cgggcttttg agcacggcta gtcgcggcgg ggggagggga
     1921 tgtaatggcg ttggagtttg ttcacatttg gtgggtggag actagtcagg ccagcctggc
     1981 gctggaagtc atttttggaa tttgtcccct tgagttttga gcggagctaa ttctcgggct
     2041 tcttagcggt tcaaaggtat cttttaaacc cttttttagg tgttgtgaaa accaccgcta
     2101 attcaaatca acaactttgt acaaaaaagt tgaacgagaa acgtaaaatg atataaatat
     2161 caatatatta aattagattt tgcataaaaa acagactaca taatactgta aaacacaaca
     2221 tatgcagtca ctatgaacca actacttaga tggtattagt gacctgtaga attcgagctc
     2281 tagagctgca gggcggccgc gatatcccct atagtgagtc gtattacatg gtcatagctg
     2341 tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa
     2401 atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta
     2461 tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt
     2521 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct
     2581 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
     2641 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
     2701 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg
     2761 gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
     2821 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
     2881 gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg
     2941 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
     3001 ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
     3061 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
     3121 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
     3181 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
     3241 ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg taacactggc
     3301 agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga
     3361 gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3421 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3481 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3541 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     3601 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3661 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3721 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3781 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3841 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3901 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3961 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     4021 cctttttacg gttcctggcc ttttgctggc cttttgctca catgtt
//
