LOCUS       sequence-122095-        1076 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Plasmid for the generation of gene disruptions via Bxb1-mediated 
            recombination into MIN-tagged cell lines. This vector can also be 
            used to express cDNAs..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1076)
  AUTHORS   Mulholland CB, Smets M, Schmidtmann E, Leidescher S, Markaki Y, 
            Hofweber M, Qin W, Manzo M, Kremmer E, Thanisch K, Bauer C, Rombaut 
            P, Herzog F, Leonhardt H, Bultmann S
  TITLE     A modular open platform for systematic functional studies under 
            physiological conditions.
  JOURNAL   Nucleic Acids Res. 2015 May 24. pii: gkv550.
  PUBMED    26007658
REFERENCE   2  (bases 1 to 1076)
  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: "Nucleic 
            Acids Res. 2015 May 24. pii: gkv550."
COMMENT     Sequence Label: attB-­GFP-­Poly(A)
FEATURES             Location/Qualifiers
     source          1..1076
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    47..84
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attB site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     CDS             join(87..89,90..800)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(87..89,90..800)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(129..150)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(390..409)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     737..758
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      846..855
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    908..1042
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(932..951)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     986..1005
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 ggatctcaag aagatccttt gatcttttct agtgtgcgga tatcccggct tgtcgacgac
       61 ggcggtctcc gtcgtcagga tcatccgtga gcaagggcga ggagctgttc accggggtgg
      121 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
      181 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
      241 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
      301 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
      361 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
      421 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
      481 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
      541 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
      601 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
      661 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
      721 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
      781 gcatggacga gctgtacaag agcggcctga ggagcagagc ccaggcgagc aacagcgccg
      841 tggacgccac catgggcgat cgcccgggaa ttgactagtg cggccgcagt taacgaattc
      901 tctagaggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
      961 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     1021 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcagg
//
