LOCUS       sequence-205359-        3235 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Knockout cassette Blasticidin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3235)
  AUTHORS   Paschke P, Knecht DA, Silale A, Traynor D, Williams TD, Thomason PA,
            Insall RH, Chubb JR, Kay RR, Veltman DM
  TITLE     Rapid and efficient genetic engineering of both wild type and axenic
            strains of Dictyostelium discoideum.
  JOURNAL   PLoS One. 2018 May 30;13(5):e0196809. doi: 
            10.1371/journal.pone.0196809. eCollection 2018.
  PUBMED    29847546
REFERENCE   2  (bases 1 to 3235)
  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: "2018-05-30"; volume: "13(5):e0196809. doi"; pages: " 
            10.1371/journal.pone.0196809. eCollection 2018"
COMMENT     Sequence Label: pDM1079
FEATURES             Location/Qualifiers
     source          1..3235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(51..84)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(1186..1219)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1305..1409
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1410..1478,1479..2270)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1628..1647)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2441..3029
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2930..2949
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3183..3200
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gccggccttc agtaggcaga gctatcaagc ttgatatctt aattaattaa ataacttcgt
       61 atagcataca ttatacgaag ttatagtagt ggatcccaaa attttgtatt ttgtgatgtt
      121 agtgtgtgtc ataattggaa tattgaaaaa aaaaaaataa aaaataaaaa aaaaaaaaaa
      181 aagaattaat taatgattaa tcatatgact aacattatat tttcaatttt tttttttttt
      241 tttttccaac cacccaaaag tgatccattt tttttttttt tttaaataaa atttgggtca
      301 cccacagttg aattattttt aatttttttt tttttttttt ttttttttaa tttttttttt
      361 ttttttcaaa attttttttt cttcatataa aaataaattt tccaattcat tttttttttt
      421 tttattcaac tgtatattat tataactctt tatttgtatt ttaaactaat ttcacagcta
      481 gcaaaaaatg gatcaattta acatttctca acaggatcta gaattagtag aagtagcgac
      541 agagaagatt acaatgcttt atgaggataa taaacatcat gtgggagcgg caattcgtac
      601 gaaaacagga gaaatcattt cggcagtaca tattgaagcg tatataggac gagtaactgt
      661 ttgtgcagaa gccattgcga ttggtagtgc agtttcgaat ggacaaaagg attttgacac
      721 gattgtagct gttagacacc cttattctga cgaagtagat agaagtattc gagtggtaag
      781 tccttgtggt atgtgtaggg agttgatttc agactatgca ccagattgtt ttgtgttaat
      841 agaaatgaat ggcaagttag tcaaaactac gattgaagaa ctcattccac tcaaatatac
      901 ccgaaattaa gcggccgcat caatttgatt tcttcttaat tcaatgctaa aaaaaaaaat
      961 attataaaaa aaaaaacaca cacaacaaat aaattaataa ataccattta ttaaaaaaaa
     1021 aaaaaaaaaa aaaaaaccta ttttaacaat tattttatta tttttatttt tagttttgtt
     1081 aaaaaaaaaa ttattatctt tatttataat tataaaaaat ttatccaatg atcatttttt
     1141 aactttttta ttatttagta tattaaataa aatctcgaga gtagtataac ttcgtatagc
     1201 atacattata cgaagttatg atatcagatc tctactggag tatccaagct gactagttga
     1261 tcccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     1321 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     1381 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     1441 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     1501 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     1561 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     1621 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     1681 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     1741 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     1801 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     1861 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     1921 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     1981 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     2041 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     2101 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     2161 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     2221 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     2281 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     2341 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     2401 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     2461 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     2521 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     2581 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2641 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2701 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2761 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2821 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2881 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2941 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3001 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3061 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     3121 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     3181 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctct
//
