LOCUS       sequence-340172-        2759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB4/attB5 entry clone for mammalian expression using the 
            human PGK promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2759)
  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 2759)
  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-E17: attB4-PGKp>-attB5
FEATURES             Location/Qualifiers
     source          1..2759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(73..159)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    207..302
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        308..807
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(343..366)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     740..759
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     protein_bind    complement(813..908)
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1027..1046)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1038..1847
                     /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(1105..1124)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1990..2578
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2479..2498
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2713..2740)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc
       61 cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg
      121 aaaggcccag tcttccgact gagcctttcg ttttatttga tgcctggcag ttccctactc
      181 tcgcgttaac gctaccatgg agctccaaat aatgatttta ttttgactga tagtgacctg
      241 ttcgttgcaa caaattgata agcaatgctt ttttataatg ccaactttgt atagaaaagt
      301 tggcaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc
      361 ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc
      421 ggtaggcgcc aaccggctcc gttctttggt ggccccttcg cgccaccttc tactcctccc
      481 ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa
      541 gtagcacgtc tcactagtct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta
      601 ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc tcagaggctg
      661 ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc gggcgcccga
      721 aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc
      781 tcctcttcct catctccggg cctttcgacc gccaactttt gtatacaaag ttggcattat
      841 aaaaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat
      901 cattatttgg agctctagag ctgcagggcg cagggcccgt gtctcaaaat ctctgatgtt
      961 acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca
     1021 gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt
     1081 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag
     1141 gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg
     1201 gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg
     1261 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac
     1321 tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag
     1381 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt
     1441 gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga
     1501 ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac
     1561 aagtctggaa agaaatgcat aaacttttgc cattctcgcc ggattcagtc gtcactcatg
     1621 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg
     1681 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg
     1741 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg
     1801 atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta
     1861 attggttgta acaccggcag agcattacgc tgacttgacg ggacggcgca agctcatgac
     1921 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1981 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     2041 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     2101 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     2161 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     2221 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     2281 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     2341 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2401 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2461 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2521 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2581 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     2641 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagcc aggaagagtt
     2701 tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg atgcctggc
//
