LOCUS       sequence-205377-        5380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  safe haven knock-in plasmid for the act5 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5380)
  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 5380)
  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: pDM1501
FEATURES             Location/Qualifiers
     source          1..5380
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    760..793
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1069..2106)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPDRE
                     MGEAN"
     protein_bind    2543..2576
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(3438..3455)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3609..4197)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3689..3708)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4368..5159,5160..5228))
                     /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     4991..5010
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5229..5333)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 actagcgccg gcattacatg tccatttatt agaaggattc aatcaaaaat taacaccagg
       61 tatattacca gatggcatta ataatttata ttctttaata ttggaaaaag aaaaaaataa
      121 ataaacaatc aatcagaaaa gataattgaa aaaaaaaaaa aagaaaatgt ttaataagaa
      181 ttaatttgga agtaaaataa tttttggatg agtaaatatt aataaactca caattaaaat
      241 ttagaatcca aaaaaatgaa aaaaaaaaaa atttttaaaa cagaatggtt gcattcattt
      301 tttttttttt ttttttatat cttttgggta tttttgggtg ttctatttat ctggataaaa
      361 aataaaatgg aatatagttt tatatatata tttttttcgt atcaagcttg aatgttttta
      421 tcaataatat gattgttggt tgctttattt tatttgaaaa aaaaattaat ttttaaaata
      481 tctttttttt cattttattt gaaaaaaaaa aaaaaaaaaa atggttttga aaatttttaa
      541 aaaaatggtt gggtcaccca caattcttat aaaaaaaagc catttggata gttttttttt
      601 tttttttttt tatttttttt attttttttt ttttgagatt tggtataaat acaaaaaaaa
      661 aattgttttt aaatcattat taataaaaac ttaaaataaa ttatatataa aaaaagatct
      721 agtactagta taaaataaat aaattgtgga tctgatatca taacttcgta taatgtatgc
      781 tatacgaagt tatactactc tcgagatttt atttaatata ctaaataata aaaaagttaa
      841 aaaatgatca ttggataaat tttttataat tataaataaa gataataatt ttttttttaa
      901 caaaactaaa aataaaaata ataaaataat tgttaaaata ggtttttttt tttttttttt
      961 ttttttaata aatggtattt attaatttat ttgttgtgtg tgtttttttt tttataatat
     1021 tttttttttt agcattgaat taagaagaaa tcaaattgat gcggccgctt agttagcctc
     1081 ccccatctcc cgatccggac gagtgctggg gcgtcggttt ccactatcgg cgagtacttc
     1141 tacacagcca tcggtccaga cggccgcgct tctgcgggcg atttgtgtac gcccgacagt
     1201 cccggctccg gatcggacga ttgcgtcgca tcgaccctgc gcccaagctg catcatcgaa
     1261 attgccgtca accaagctct gatagagttg gtcaagacca atgcggagca tatacgcccg
     1321 gagccgcggc gatcctgcaa gctccggatg cctccgctcg aagtagcgcg tctgctgctc
     1381 catacaagcc aaccacggcc tccagaagaa gatgttggcg acctcgtatt gggaatcccc
     1441 gaacatcgcc tcgctccagt caatgaccgc tgttatgcgg ccattgtccg tcaggacatt
     1501 gttggagccg aaatccgcgt gcacgaggtg ccggacttcg gggcagtcct cggcccaaag
     1561 catcagctca tcgagagcct gcgcgacgga cgcactgacg gtgtcgtcca tcacagtttg
     1621 ccagtgatac acatggggat cagcaatcgc gcatatgaaa tcacgccatg tagtgtattg
     1681 accgattcct tgcggtccga atgggccgaa cccgctcgtc tggctaagat cggccgcagc
     1741 gatcgcatcc atggcctccg cgaccggctg cagaacagcg ggcagttcgg tttcaggcag
     1801 gtcttgcaac gtgacaccct gtgcacggcg ggagatgcaa taggtcaggc tctcgctgaa
     1861 ctccccaatg tcaagcactt ccggaatcgg gagcgcggcc gatgcaaagt gccgataaac
     1921 ataacgatct ttgtagaaac catcggcgca gctatttacc cgcaggacat atccacgccc
     1981 tcctacatcg aagctgaaag cacgagattc ttcgccctcc gagagctgca tcaggtcgga
     2041 gacgctgtcg aacttttcga tcagaaactt ctcgacagac gtcgcggtga gttcaggctt
     2101 tttcattagg tcttgttgag aaatgttaaa ttgatccatt ttttgctagc tgtgaaatta
     2161 gtttaaaata caaataaaga gttataataa tatacagttg aataaaaaaa aaaaaaatga
     2221 attggaaaat ttatttttat atgaagaaaa aaaaattttg aaaaaaaaaa aaaaaattaa
     2281 aaaaaaaaaa aaaaaaaaaa aaaattaaaa ataattcaac tgtgggtgac ccaaatttta
     2341 tttaaaaaaa aaaaaaaaat ggatcacttt tgggtggttg gaaaaaaaaa aaaaaaaaaa
     2401 ttgaaaatat aatgttagtc atatgattaa tcattaatta attctttttt tttttttttt
     2461 tattttttat tttttttttt tcaatattcc aattatgaca cacactaaca tcacaaaata
     2521 caaaattttg ggatccacta ctataacttc gtataatgta tgctatacga agttatttaa
     2581 ttaattaaga tatcaagctt gtccacagaa aatgtttcta aacaaaaaaa aaaaaccgag
     2641 tgatgaaagc gcttctcaca aaattattta tgtaaaatat ttaataaaat acattattta
     2701 aatctttttt atttttttta gttgttgatc tttatccgac tttaaaaata aaaaaattgt
     2761 aaaaaaaaaa aaagtttatt tgtttaattt tattgttgtt tttaattttt tacaaccatt
     2821 aatttataaa accaacacct aactataatg aaatacaatt tgtatcttgg aaaaaaaaaa
     2881 attaataata taaataaaat gtaaacaagt atccggtgga aggaactaac agttgcctta
     2941 tccagttcaa aaaaaaaaaa aaaaaaagaa attaaattac cattttatat ttaactaaaa
     3001 taaattaaaa ggtttatttt aataatagat ttattatttt ttaaaaatca ataattaatg
     3061 ttggataaga tgaacgaata aaagatagag tttcaggtct tttaaatgat gaggatttat
     3121 gaattcttaa aagttttggg caagagaaaa tgattttaac gattgaacca tcactaattg
     3181 caaaatctct tgaactaaga gtttttaagt ttgaagcaag tggagaatta caaagagcaa
     3241 tcatagagtt atcagttaaa tcataacaac aagataaaga taaagaacta atatgatgta
     3301 aattttcaat catacctcta aaagaaacat cattcatact ttttggataa attaaatcta
     3361 atgaatctaa atttggaaat agtaaaccaa tatttggccg gcagaggcgg tttgcgtatt
     3421 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3481 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3541 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3601 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3661 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3721 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3781 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3841 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3901 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3961 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4021 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     4081 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4141 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4201 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4261 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4321 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4381 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4441 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4501 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4561 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4621 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4681 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     4741 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     4801 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     4861 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     4921 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     4981 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5041 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5101 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5161 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5221 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5281 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5341 ccccgaaaag tgccacctga cgcgccctgt agcgggatca
//
