LOCUS       sequence-216056-        3881 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Knockout cassette Hygromycin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3881)
  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 3881)
  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: pDM1081
FEATURES             Location/Qualifiers
     source          1..3881
                     /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)."
     CDS             520..1557
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPDRE
                     MGEAN"
     protein_bind    complement(1832..1865)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1951..2055
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2056..2124,2125..2916)
                     /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(2274..2293)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3087..3675
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3576..3595
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3829..3846
                     /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 tttttttawt tttttttttt
      361 tttttyaaaa tttttttttt ttyatataaa aataaatttt ccaattcatt tttttttttt
      421 ttattcaact gtatattatt ataactcttt atttgtattt taaactaatt tcacagctag
      481 caaaaaatgg atcaatttaa catttctcaa caagacctaa tgaaaaagcc tgaactcacc
      541 gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag
      601 ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc
      661 ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt
      721 gcatcggccg cgctcccgat tccggaagtg cttgacattg gggagttcag cgagagcctg
      781 acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa
      841 ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc ggccgatctt
      901 agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg
      961 cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac
     1021 gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac
     1081 tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac
     1141 aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac
     1201 gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc
     1261 tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc gtatatgctc
     1321 cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct
     1381 tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca
     1441 caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt actcgccgat
     1501 agtggaaacc gacgccccag cactcgtccg gatcgggaga tgggggaggc taactaagcg
     1561 gccgcatcaa tttgatttct tcttaattca atgctaaaaa aaaaaatatt ataaaaaaaa
     1621 aacacacaca acaaataaat taataaatac catttattaa aaaaaaaaaa aaaaaaaaaa
     1681 aacctatttt aacaattatt ttattatttt tatttttagt tttgttaaaa aaaaaattat
     1741 tatctttatt tataattata aaaaatttat ccaatgatca ttttttaact tttttattat
     1801 ttagtatatt aaataaaatc tcgagagtag tataacttcg tatagcatac attatacgaa
     1861 gttatgatat cagatctcta ctggagtatc caagctgact agttgatccc gctacagggc
     1921 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     1981 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     2041 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     2101 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     2161 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     2221 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     2281 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     2341 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     2401 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     2461 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     2521 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     2581 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     2641 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     2701 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     2761 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     2821 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     2881 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     2941 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     3001 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     3061 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3121 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3181 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     3241 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     3301 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     3361 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     3421 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     3481 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     3541 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     3601 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     3661 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     3721 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3781 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     3841 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc t
//
