LOCUS       sequence-8511-ex        2208 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 2208)
  AUTHORS   Kohli M, Rago C, Lengauer C, Kinzler KW, Vogelstein B
  TITLE     Facile methods for generating human somatic cell gene knockouts 
            using recombinant adeno-associated viruses.
  JOURNAL   Nucleic Acids Res. 2004  . 32(1):e3.
  PUBMED    14704360
REFERENCE   2  (bases 1 to 2208)
  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. 2004  . 32(1):e3."
COMMENT     Sequence Label: pNeDaKO-Neo
FEATURES             Location/Qualifiers
     source          1..2208
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    complement(34..67)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        79..579
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(114..137)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     512..531
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             593..1396
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVNQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(656..675)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1266..1285
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1428..1445)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1434..1658
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        1699..1746
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        1738..1756
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1760..1769
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1766..2140
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     protein_bind    complement(2141..2174)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(2189..2207)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ctaaagggaa caaaagctgg agctccaccg cggataactt cgtatagcat acattatacg
       61 aagttatcgc gccctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg
      121 ctttagcagc cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt
      181 ccacatccac cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt
      241 ctactcctcc cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt
      301 gacaaatgga agtagcacgt ctcactagtc tcgtgcagat ggacagcacc gctgagcaat
      361 ggaagcgggt aggcctttgg ggcagcggcc aatagcagct ttggctcctt cgctttctgg
      421 gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct caggggcggg
      481 gcgggcgccc gaaggtcctc cggaagcccg gcattctgca cgcttcaaaa gcgcacgtct
      541 gccgcgctgt tctcctcttc ctcatctccg ggcctttcga cctgcagcca atatgggatc
      601 ggccattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
      661 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
      721 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
      781 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
      841 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
      901 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
      961 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     1021 catcgagcga gcacgtactc ggatggaagc cggtcttgtc aatcaggatg atctggacga
     1081 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     1141 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     1201 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggatc gctatcagga
     1261 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     1321 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     1381 tgacgagttc ttctgagggg atcaattctc tagagctcgc tgatcagcct cgactgtgcc
     1441 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     1501 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     1561 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     1621 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     1681 ctgggggctg cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     1741 tacgactcac tataggaggg ccaccatggc caagttgacc agtgccgttc cggtgctcac
     1801 cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga
     1861 cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc
     1921 ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga
     1981 cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc
     2041 ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc
     2101 cggcaactgc gtgcacttcg tggccgagga gcaggactga ataacttcgt atagcataca
     2161 ttatacgaag ttatggtacc caattcgccc tatagtgagt cgtattac
//
