LOCUS       sequence-70474-e         874 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 874)
  AUTHORS   Dolt KS, Lawrence ML, Miller-Hodges E, Slight J, Thornburn A, 
            Devenney PS, Hohenstein P
  TITLE     A universal vector for high-efficiency multi-fragment recombineering
            of BACs and knock-in constructs.
  JOURNAL   PLoS One. 2013 Apr 29;8(4):e62054. doi: 
            10.1371/journal.pone.0062054. Print 2013.
  PUBMED    23637962
REFERENCE   2  (bases 1 to 874)
  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: "PLoS One.";
            date: "2013-04-29"; volume: "8(4):e62054. doi"; pages: " 
            10.1371/journal.pone.0062054. Print 2013"
COMMENT     Sequence Label: pMULTIrec
FEATURES             Location/Qualifiers
     source          1..874
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     68..84
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     69..85
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    101..125
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(131..133,134..136,137..847)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(131..133,134..136,137..847)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(176..197)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(437..456)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     784..805
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 tatagggcga attgggtacc gggccccccc tcgaagatct catatggttt aaacacgcgt
       61 gctagcatcg aggtcgacgg tatcgataag cttgatatca acaagtttgt acaaaaaagc
      121 aggcttaacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
      181 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
      241 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
      301 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
      361 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
      421 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
      481 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
      541 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
      601 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
      661 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
      721 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
      781 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
      841 gtacaagtcc ggtacagctc tcgacggaga aagc
//
