LOCUS       sequence-330927-        3535 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB2r/attB3 entry clone for construction of C-terminal 
            mVenus fusion proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3535)
  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 3535)
  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: C232-E25: attB2r-mVenus-attB3
FEATURES             Location/Qualifiers
     source          1..3535
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..45
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     37..54
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     38..54
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(152..275)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(278..280,281..994)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(320..341)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(581..600)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     928..949
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(996..1091)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1502..2293
                     /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      2386..2974
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2875..2894
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3128..3145
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3304..3331)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3423..3509)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaaa
       61 attcgagctc ggtccgatat ccttaagctc gggccctgca gctctagagc tcgaattcta
      121 caggtcacta ataccatcta agtagttggt tcatagtgac tgcatatgtt gtgttttaca
      181 gtattatgta gtctgttttt tatgcaaaat ctaatttaat atattgatat ttatatcatt
      241 ttacgtttct cgttcaactt tcttgtacaa agtgggcgtg agcaagggcg aggagctgtt
      301 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
      361 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctgatctg
      421 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcct
      481 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
      541 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
      601 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
      661 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
      721 caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg
      781 ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat
      841 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccaagctgag
      901 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
      961 gatcactctc ggcatggacg agctgtacaa gtaatcaact ttattataca aagttggcat
     1021 tataaaaaag cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa
     1081 aatcattatt tggagctcca tggtagcgtt aacgcggccg cgatatcggt acccggggat
     1141 cctctagacc agccaggaca gaaatgcctc gacttcgctg ctacccaagg ttgccgggtg
     1201 acgcacaccg tggaaacgga tgaaggcacg aacccagtgg acataagcct gttcggttcg
     1261 taagctgtaa tgcaagtagc gtatgcgctc acgcaactgg tccagaacct tgaccgaacg
     1321 cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttggggt
     1381 acagtctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat
     1441 gttatggagc agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaaacat
     1501 tatgagggaa gcggtgatcg ccgaagtatc gactcaacta tcagaggtag ttggcgtcat
     1561 cgagcgccat ctcgaaccga cgttgctggc cgtacatttg tacggctccg cagtggatgg
     1621 cggcctgaag ccacacagtg atattgattt gctggttacg gtgaccgtaa ggcttgatga
     1681 aacaacgcgg cgagctttga tcaacgacct tttggaaact tcggcttccc ctggagagag
     1741 cgagattctc cgcgctgtag aagtcaccat tgttgtgcac gacgacatca ttccgtggcg
     1801 ttatccagct aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg
     1861 tatcttcgag ccagccacga tcgacattga tctggctatc ttgctgacaa aagcaagaga
     1921 acatagcgtt gccttggtag gtccagcggc ggaggaactc tttgatccgg ttcctgaaca
     1981 ggatctattt gaggcgctaa atgaaacctt aacgctatgg aactcgccgc ccgactgggc
     2041 tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg
     2101 caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc cggcccagta
     2161 tcagcccgtc atacttgaag ctagacaggc ttatcttgga caagaagaag atcgcttggc
     2221 ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt
     2281 agtcggcaaa taaccctcga gccacccatg accaaaatcc cttaacgtga gttacgcgtc
     2341 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     2401 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     2461 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     2521 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     2581 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     2641 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     2701 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     2761 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     2821 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     2881 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     2941 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     3001 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     3061 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     3121 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     3181 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     3241 gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc caggaagagt ttgtagaaac
     3301 gcaaaaaggc catccgtcag gatggccttc tgcttagttt gatgcctggc agtttatggc
     3361 gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc
     3421 ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag
     3481 tcttccgact gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcg
//
