LOCUS       sequence-216071-        5374 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 5374)
  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 5374)
  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..5374
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    757..790
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1064..2101)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPDRE
                     MGEAN"
     protein_bind    2537..2570
                     /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(3432..3449)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3603..4191)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3683..3702)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4362..5153,5154..5222))
                     /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     4985..5004
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5223..5327)
                     /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 ttttatatct tttgggtatt tttgggtgtt ctatttatct ggataaaaaa
      361 taaaatggaa tatagtttta tatatatatt tttttcgtat caagcttgaa tgtttttatc
      421 aataatatga ttgttggttg ctttatttta tttgaaaaaa aaattaattt ttaaaatatc
      481 ttttttttca ttttatttga aaaaaaaaaa aaaaaaaatg gttttgaaaa tttttaaaaa
      541 aatggttggg tcacccamaa ttyttataaa aaaaarccat ttggataktt tttttttttt
      601 ttttttttat tttttttatt tttttttttt tgagatttgg tataaataca aaaaaaaaat
      661 tgtttttaaa tcattattaa taaaaactta aaataaatta tatataaaaa aagatctagt
      721 actagtataa aataaataaa ttgtggatct gatatcataa cttcgtataa tgtatgctat
      781 acgaagttat actactctcg agattttatt taatatacta aataataaaa aagttaaaaa
      841 atgatcattg gataaatttt ttataattat aaataaagat aataattttt tttttaacaa
      901 aactaaaaat aaaaataata aaataattgt taaaataggt tttttttttt tttttttttt
      961 ttaataaatg gtatttatta atttatttgt tgtgtgtgtt ttttttttat aatatttttt
     1021 tttttagcat tgaattaaga agaaatcaaa ttgatgcggc cgcttagtta gcctccccca
     1081 tctcccgatc cggacgagtg ctggggcgtc ggtttccact atcggcgagt acttctacac
     1141 agccatcggt ccagacggcc gcgcttctgc gggcgatttg tgtacgcccg acagtcccgg
     1201 ctccggatcg gacgattgcg tcgcatcgac cctgcgccca agctgcatca tcgaaattgc
     1261 cgtcaaccaa gctctgatag agttggtcaa gaccaatgcg gagcatatac gcccggagcc
     1321 gcggcgatcc tgcaagctcc ggatgcctcc gctcgaagta gcgcgtctgc tgctccatac
     1381 aagccaacca cggcctccag aagaagatgt tggcgacctc gtattgggaa tccccgaaca
     1441 tcgcctcgct ccagtcaatg accgctgtta tgcggccatt gtccgtcagg acattgttgg
     1501 agccgaaatc cgcgtgcacg aggtgccgga cttcggggca gtcctcggcc caaagcatca
     1561 gctcatcgag agcctgcgcg acggacgcac tgacggtgtc gtccatcaca gtttgccagt
     1621 gatacacatg gggatcagca atcgcgcata tgaaatcacg ccatgtagtg tattgaccga
     1681 ttccttgcgg tccgaatggg ccgaacccgc tcgtctggct aagatcggcc gcagcgatcg
     1741 catccatggc ctccgcgacc ggctgcagaa cagcgggcag ttcggtttca ggcaggtctt
     1801 gcaacgtgac accctgtgca cggcgggaga tgcaataggt caggctctcg ctgaactccc
     1861 caatgtcaag cacttccgga atcgggagcg cggccgatgc aaagtgccga taaacataac
     1921 gatctttgta gaaaccatcg gcgcagctat ttacccgcag gacatatcca cgccctccta
     1981 catcgaagct gaaagcacga gattcttcgc cctccgagag ctgcatcagg tcggagacgc
     2041 tgtcgaactt ttcgatcaga aacttctcga cagacgtcgc ggtgagttca ggctttttca
     2101 ttaggtcttg ttgagaaatg ttaaattgat ccattttttg ctagctgtga aattagttta
     2161 aaatacaaat aaagagttat aataatatac agttgaataa aaaaaaaaaa aatraattgg
     2221 aaaatttatt tttatatraa aaaaaaaaaa ttttraaaaa aaaaaaaaaa awwaaaaaaa
     2281 aaaaaaaaaa aaaaaaaaat taaaaawaat tcaactgtgg gtgacccaaa ttttatttaa
     2341 aaaaaaaaaa aaaatggatc acttttgggt ggttggaaaa aaaaaaaaaa aaaaattgaa
     2401 aatataatgt tagtcatatg attaatcatt aattaattct tttttttttt ttttttattt
     2461 tttatttttt tttttcaata ttccaattat gacacacact aacatcacaa aatacaaaat
     2521 tttgggatcc actactataa cttcgtataa tgtatgctat acgaagttat ttaattaatt
     2581 aagatatcaa gcttgtccac agaaaatgtt tctaaacaaa aaaaaaaaac cgagtgatga
     2641 aagcgcttct cacaaaatta tttatgtaaa atatttaata aaatacatta tttaaatctt
     2701 ttttattttt tttagttgtt gatctttatc cgactttaaa aataaaaaaa ttgtaaaaaa
     2761 aaaaaaagtt tatttgttta attttattgt tgtttttaat tttttacaac cattaattta
     2821 taaaaccaac acctaactat aatgaaatac aatttgtatc ttggaaaaaa aaaaattaat
     2881 aatataaata aaatgtaaac aagtatccgg tggaaggaac taacagttgc cttatccagt
     2941 tcaaaaaaaa aaaaaaaaaa agaaattaaa ttaccatttt atatttaact aaaataaatt
     3001 aaaaggttta ttttaataat agatttatta ttttttaaaa atcaataatt aatgttggat
     3061 aagatgaacg aataaaagat agagtttcag gtcttttaaa tgatgaggat ttatgaattc
     3121 ttaaaagttt tgggcaagag aaaatgattt taacgattga accatcacta attgcaaaat
     3181 ctcttgaact aagagttttt aagtttgaag caagtggaga attacaaaga gcaatcatag
     3241 agttatcagt taaatcataa caacaagata aagataaaga actaatatga tgtaaatttt
     3301 caatcatacc tctaaaagaa acatcattca tactttttgg ataaattaaa tctaatgaat
     3361 ctaaatttgg aaatagtaaa ccaatatttg gccggcagag gcggtttgcg tattgggcgc
     3421 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3481 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3541 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3601 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3661 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3721 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3781 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3841 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3901 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3961 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4021 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4081 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4141 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4201 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4261 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4321 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4381 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4441 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4501 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4561 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4621 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     4681 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     4741 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     4801 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     4861 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     4921 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     4981 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5041 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5101 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5161 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5221 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5281 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5341 aaagtgccac ctgacgcgcc ctgtagcggg atca
//
