LOCUS       sequence-205361-        3883 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 3883)
  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 3883)
  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..3883
                     /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             521..1558
                     /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(1834..1867)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1953..2057
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2058..2126,2127..2918)
                     /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(2276..2295)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3089..3677
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3578..3597
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3831..3848
                     /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 acaagaccta atgaaaaagc ctgaactcac
      541 cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca
      601 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt
      661 cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt
      721 tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggagttca gcgagagcct
      781 gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga
      841 actgcccgct gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct
      901 tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg
      961 gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga
     1021 cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga
     1081 ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga
     1141 caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata
     1201 cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg
     1261 ctacttcgag cggaggcatc cggagcttgc aggatcgccg cggctccggg cgtatatgct
     1321 ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc
     1381 ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac
     1441 acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga
     1501 tagtggaaac cgacgcccca gcactcgtcc ggatcgggag atgggggagg ctaactaagc
     1561 ggccgcatca atttgatttc ttcttaattc aatgctaaaa aaaaaaatat tataaaaaaa
     1621 aaaacacaca caacaaataa attaataaat accatttatt aaaaaaaaaa aaaaaaaaaa
     1681 aaaacctatt ttaacaatta ttttattatt tttattttta gttttgttaa aaaaaaaatt
     1741 attatcttta tttataatta taaaaaattt atccaatgat cattttttaa cttttttatt
     1801 atttagtata ttaaataaaa tctcgagagt agtataactt cgtatagcat acattatacg
     1861 aagttatgat atcagatctc tactggagta tccaagctga ctagttgatc ccgctacagg
     1921 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     1981 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2041 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2101 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2161 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2221 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     2281 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     2341 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     2401 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     2461 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2521 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2581 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2641 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     2701 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     2761 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     2821 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     2881 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     2941 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3001 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3061 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3121 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3181 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     3241 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3301 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3361 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3421 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3481 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3541 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3601 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3661 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3721 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     3781 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     3841 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tct
//
