LOCUS       sequence-205360-        3337 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Knockout cassette Nourseothricin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3337)
  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 3337)
  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: pDM1080
FEATURES             Location/Qualifiers
     source          1..3337
                     /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(1288..1321)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1407..1511
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1512..1580,1581..2372)
                     /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(1730..1749)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2543..3131
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3032..3051
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3285..3302
                     /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 aaaatttcag ttattccaga acaagttgca gaaacattag atgcagaaaa
      541 tcattttatt gttagagaag tttttgatgt tcatttatca gatcaaggtt ttgaattatc
      601 aacaagatca gtttcaccat atagaaaaga ttatatttca gatgatgatt cagatgaaga
      661 ttcagcatgt tatggtgcat ttattgatca agaattagtt ggtaaaattg aattaaattc
      721 aacatggaat gatttagcat caattgaaca tattgttgtt tcacatacac atagaggtaa
      781 aggtgttgca cattcattaa ttgaatttgc aaaaaaatgg gcattatcaa gacaattatt
      841 aggtattaga ttagaaacac aaacaaataa tgttccagca tgtaatttat atgcaaaatg
      901 tggttttaca ttaggtggta ttgatttatt tacatataaa acaagaccac aagtttcaaa
      961 tgaaacagca atgtattggt attggttttc aggtgcacaa gatgatgcat aagcggccgc
     1021 atcaatttga tttcttctta attcaatgct aaaaaaaaaa atattataaa aaaaaaaaca
     1081 cacacaacaa ataaattaat aaataccatt tattaaaaaa aaaaaaaaaa aaaaaaaacc
     1141 tattttaaca attattttat tatttttatt tttagttttg ttaaaaaaaa aattattatc
     1201 tttatttata attataaaaa atttatccaa tgatcatttt ttaacttttt tattatttag
     1261 tatattaaat aaaatctcga gagtagtata acttcgtata gcatacatta tacgaagtta
     1321 tgatatcaga tctctactgg agtatccaag ctgactagtt gatcccgcta cagggcgcgt
     1381 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     1441 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     1501 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     1561 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     1621 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     1681 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     1741 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     1801 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     1861 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     1921 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     1981 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     2041 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     2101 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     2161 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     2221 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     2281 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     2341 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     2401 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     2461 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     2521 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2581 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2641 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     2701 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2761 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     2821 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     2881 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     2941 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3001 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     3061 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3121 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3181 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     3241 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     3301 aggaagcgga agagcgccca atacgcaaac cgcctct
//
