LOCUS       sequence-216058-        3334 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 3334)
  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 3334)
  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..3334
                     /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(1285..1318)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1404..1508
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1509..1577,1578..2369)
                     /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(1727..1746)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2540..3128
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3029..3048
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3282..3299
                     /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 agaattaatt aatgattaat catatgacta acattatatt ttcaattttt tttttttttt
      241 ttttccaacc acccaaaagt gatccatttt tttttttttt ttaaataaaa tttgggtcac
      301 ccacagttga attwttttta attttttttt tttttttttt tttttttwwt tttttttttt
      361 tttttyaaaa tttttttttt ttyatataaa aataaatttt ccaattcatt tttttttttt
      421 ttattcaact gtatattatt ataactcttt atttgtattt taaactaatt tcacagctag
      481 caaaaaatga aaatttcagt tattccagaa caagttgcag aaacattaga tgcagaaaat
      541 cattttattg ttagagaagt ttttgatgtt catttatcag atcaaggttt tgaattatca
      601 acaagatcag tttcaccata tagaaaagat tatatttcag atgatgattc agatgaagat
      661 tcagcatgtt atggtgcatt tattgatcaa gaattagttg gtaaaattga attaaattca
      721 acatggaatg atttagcatc aattgaacat attgttgttt cacatacaca tagaggtaaa
      781 ggtgttgcac attcattaat tgaatttgca aaaaaatggg cattatcaag acaattatta
      841 ggtattagat tagaaacaca aacaaataat gttccagcat gtaatttata tgcaaaatgt
      901 ggttttacat taggtggtat tgatttattt acatataaaa caagaccaca agtttcaaat
      961 gaaacagcaa tgtattggta ttggttttca ggtgcacaag atgatgcata agcggccgca
     1021 tcaatttgat ttcttcttaa ttcaatgcta aaaaaaaaaa tattataaaa aaaaaacaca
     1081 cacaacaaat aaattaataa ataccattta ttaaaaaaaa aaaaaaaaaa aaaaacctat
     1141 tttaacaatt attttattat ttttattttt agttttgtta aaaaaaaaat tattatcttt
     1201 atttataatt ataaaaaatt tatccaatga tcatttttta acttttttat tatttagtat
     1261 attaaataaa atctcgagag tagtataact tcgtatagca tacattatac gaagttatga
     1321 tatcagatct ctactggagt atccaagctg actagttgat cccgctacag ggcgcgtcag
     1381 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     1441 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     1501 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     1561 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     1621 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     1681 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     1741 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     1801 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     1861 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     1921 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     1981 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     2041 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     2101 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     2161 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     2221 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     2281 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     2341 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     2401 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     2461 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     2521 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     2581 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     2641 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     2701 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     2761 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     2821 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     2881 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     2941 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3001 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     3061 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3121 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     3181 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     3241 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     3301 aagcggaaga gcgcccaata cgcaaaccgc ctct
//
