LOCUS       sequence-330544-        3403 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB5r/attB1r entry clone for generation of N-terminal 
            mVenus fusion proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3403)
  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 3403)
  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: C512-E15: attB5r-mVenus-nostop-attB1r
FEATURES             Location/Qualifiers
     source          1..3403
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(44..167)
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(172..174,175..177,178..888)
                     /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="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(217..238)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(478..497)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     825..846
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    889..1013
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1085..1104)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1086..1104)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1109..1125)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1109..1125)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1550..2341
                     /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      2434..3022
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2923..2942
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3157..3184)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3276..3362)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 agctcgaatt ctacagtcac taataccatc taagtagttg attcatagtg actgcatatg
       61 ttgtgtttta cagtattatg tagtctgttt tttatgcaaa atctaattta atatattgat
      121 atttatatca ttttacgttt ctcgttcaac ttttgtatac aaagttgaac catggtgagc
      181 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
      241 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
      301 accctgaagc tgatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
      361 accctgggct acggcctgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
      421 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
      481 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
      541 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
      601 tacaactaca acagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag
      661 gccaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac
      721 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
      781 taccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
      841 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagac aactttgtac
      901 aaaaaagttg aacgagaaac gtaaaatgat ataaatatca atatattaaa ttagattttg
      961 cataaaaaac agactacata atactgtaaa acacaacata tgcagtcact atgaaccaac
     1021 tacttagatg gtattagtga cctgtagaat tcgagctcta gagctgcagg gcggccgcga
     1081 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     1141 gtctcaaaat ctctgatgtt acattgcaca agatcggtac ccggggatcc tctagaccag
     1201 ccaggacaga aatgcctcga cttcgctgct acccaaggtt gccgggtgac gcacaccgtg
     1261 gaaacggatg aaggcacgaa cccagtggac ataagcctgt tcggttcgta agctgtaatg
     1321 caagtagcgt atgcgctcac gcaactggtc cagaaccttg accgaacgca gcggtggtaa
     1381 cggcgcagtg gcggttttca tggcttgtta tgactgtttt tttggggtac agtctatgcc
     1441 tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag
     1501 caacgatgtt acgcagcagg gcagtcgccc taaaacaaag ttaaacatta tgagggaagc
     1561 ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg agcgccatct
     1621 cgaaccgacg ttgctggccg tacatttgta cggctccgca gtggatggcg gcctgaagcc
     1681 acacagtgat attgatttgc tggttacggt gaccgtaagg cttgatgaaa caacgcggcg
     1741 agctttgatc aacgaccttt tggaaacttc ggcttcccct ggagagagcg agattctccg
     1801 cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa
     1861 gcgcgaactg caatttggag aatggcagcg caatgacatt cttgcaggta tcttcgagcc
     1921 agccacgatc gacattgatc tggctatctt gctgacaaaa gcaagagaac atagcgttgc
     1981 cttggtaggt ccagcggcgg aggaactctt tgatccggtt cctgaacagg atctatttga
     2041 ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc gactgggctg gcgatgagcg
     2101 aaatgtagtg cttacgttgt cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc
     2161 gaaggatgtc gctgccgact gggcaatgga gcgcctgccg gcccagtatc agcccgtcat
     2221 acttgaagct agacaggctt atcttggaca agaagaagat cgcttggcct cgcgcgcaga
     2281 tcagttggaa gaatttgtcc actacgtgaa aggcgagatc accaaggtag tcggcaaata
     2341 accctcgagc cacccatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     2401 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2461 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2521 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2581 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2641 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2701 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2761 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2821 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2881 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     2941 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3001 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3061 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     3121 tattaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc
     3181 ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc
     3241 cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc
     3301 accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt
     3361 tgatgcctgg cagttcccta ctctcgcgtt aacgctacca tgg
//
