LOCUS       sequence-213755-        3231 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 3231)
  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 3231)
  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..3231
                     /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(1182..1215)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1301..1405
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1406..1474,1475..2266)
                     /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(1624..1643)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2437..3025
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2926..2945
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3179..3196
                     /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 aaaaaataaa aaataaaaaa aaaaaaaaaa
      181 gaattaatta atgattaatc atatgactaa cattatattt tcaatttttt tttttttttt
      241 tttccaacca cccaaaagtg atccattttt tttttttttt taaataaaat ttgggtcacc
      301 cacakttkaa ttttttttww tttttttttt tttttttttt ttttttaatt tttttttttt
      361 ttttyaaaat tttttttttt tcatataaaa ataaattttc caattcattt tttttttttt
      421 tattcaactg tatattatta taactcttta tttgtatttt aaactaattt cacagctagc
      481 aaaaaatgga tcaatttaac atttctcaac aggatctaga attagtagaa gtagcgacag
      541 agaagattac aatgctttat gaggataata aacatcatgt gggagcggca attcgtacga
      601 aaacaggaga aatcatttcg gcagtacata ttgaagcgta tataggacga gtaactgttt
      661 gtgcagaagc cattgcgatt ggtagtgcag tttcgaatgg acaaaaggat tttgacacga
      721 ttgtagctgt tagacaccct tattctgacg aagtagatag aagtattcga gtggtaagtc
      781 cttgtggtat gtgtagggag ttgatttcag actatgcacc agattgtttt gtgttaatag
      841 aaatgaatgg caagttagtc aaaactacga ttgaagaact cattccactc aaatataccc
      901 gaaattaagc ggccgcatca atttgatttc ttcttaattc aatgctaaaa aaaaaaatat
      961 tataaaaaaa aaacacacac aacaaataaa ttaataaata ccatttatta aaaaaaaaaa
     1021 aaaaaaaaaa aacctatttt aacaattatt ttattatttt tatttttagt tttgttaaaa
     1081 aaaaaattat tatctttatt tataattata aaaaatttat ccaatgatca ttttttaact
     1141 tttttattat ttagtatatt aaataaaatc tcgagagtag tataacttcg tatagcatac
     1201 attatacgaa gttatgatat cagatctcta ctggagtatc caagctgact agttgatccc
     1261 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     1321 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     1381 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     1441 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     1501 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     1561 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     1621 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     1681 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     1741 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     1801 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     1861 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     1921 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     1981 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     2041 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     2101 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     2161 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     2221 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     2281 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     2341 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2401 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2461 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2521 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2581 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2641 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2701 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     2761 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     2821 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     2881 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     2941 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3001 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3061 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3121 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     3181 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc t
//
