LOCUS       sequence-333337-        3001 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB4/attB1r entry clone for mammalian expression using the 
            short EF1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3001)
  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 3001)
  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-E40: attB4-EFSp>-attB1r
FEATURES             Location/Qualifiers
     source          1..3001
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     48..70
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     62..79
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     63..79
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    109..204
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        232..443
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     protein_bind    458..582
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             993..1784
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      1877..2465
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2366..2385
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2619..2636
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2795..2822)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2914..3000)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tgcctggcag ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg
       61 ttgtaaaacg acggccagtc ttaaaattcg agctcggtcc gaggatccaa ataatgattt
      121 tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc ttttttataa
      181 tgccaacttt gtatagaaaa gttggctcga gtggctccgg tgcccgtcag tgggcagagc
      241 gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct
      301 agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc
      361 ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca
      421 acgggtttgc cgccagaaca caggtgtcgt gacgcggaca actttgtaca aaaaagctga
      481 acgagaaacg taaaatgata taaatatcaa tatattaaat tagattttgc ataaaaaaca
      541 gactacataa tactgtaaaa cacaacatat gcagtcacta tgaatcaact acttagatgg
      601 tattagtgac ctgtagcatg caagcttaga tcctctagac cagccaggac agaaatgcct
      661 cgacttcgct gctacccaag gttgccgggt gacgcacacc gtggaaacgg atgaaggcac
      721 gaacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag cgtatgcgct
      781 cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt
      841 tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc
      901 aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc
      961 agggcagtcg ccctaaaaca aagttaaaca ttatgaggga agcggtgatc gccgaagtat
     1021 cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg
     1081 ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt
     1141 tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc
     1201 ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca
     1261 ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg
     1321 gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg
     1381 atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg
     1441 cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct
     1501 taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt
     1561 tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg
     1621 actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg
     1681 cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg
     1741 tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg agccacccat
     1801 gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa
     1861 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1921 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1981 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     2041 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     2101 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     2161 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     2221 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     2281 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     2341 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     2401 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     2461 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     2521 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     2581 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     2641 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     2701 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatacgc
     2761 gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt
     2821 ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg
     2881 ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga gagcgttcac
     2941 cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc gttttatttg
     3001 a
//
