LOCUS       sequence-213617-        4589 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  generic knockin vector for C-terminal tagging with mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4589)
  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 4589)
  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: pPI461
FEATURES             Location/Qualifiers
     source          1..4589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    745..778
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1054..2091)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPDRE
                     MGEAN"
     protein_bind    2528..2561
                     /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(2647..2664)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2818..3406)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2898..2917)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3577..4368,4369..4437))
                     /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     4200..4219
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4438..4542)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 actagtggtg gttcaggagg tagtgtttca aaaggtgaag aagataatat ggcaattatt
       61 aaagaattta tgagatttaa agttcatatg gaaggttcag ttaatggtca tgaatttgaa
      121 attgaaggtg aaggtgaagg tagaccatat gaaggtacac aaacagcaaa attaaaagtt
      181 acaaaaggtg gtccattacc atttgcatgg gatattttat caccacaatt tatgtatggt
      241 tcaaaagcat atgttaaaca tccagcagat attccagatt atttaaaatt atcatttcca
      301 gaaggtttta aatgggaaag agttatgaat tttgaagatg gtggtgttgt tacagttaca
      361 caagattcat cattacaaga tggtgaattt atttataaag ttaaattaag aggtacaaat
      421 tttccatcag atggtccagt tatgcaaaag aaaacaatgg gttgggaagc atcatcagaa
      481 agaatgtatc cagaagatgg tgcattaaaa ggtgaaatta aacaaagatt aaaattaaaa
      541 gatggtggtc attatgatgc agaagttaaa acaacatata aagcaaaaaa accagttcaa
      601 ttaccaggtg catataatgt taatattaaa ttagatatta catcacataa tgaagattat
      661 acaattgttg aacaatatga aagagcagaa ggtagacatt caacaggtgg tatggatgaa
      721 ttatataaat aaagatctga tatcataact tcgtataatg tatgctatac gaagttatac
      781 tactctcgag attttattta atatactaaa taataaaaaa gttaaaaaat gatcattgga
      841 taaatttttt ataattataa ataaagataa taattttttt tttaacaaaa ctaaaaataa
      901 aaataataaa ataattgtta aaataggttt tttttttttt tttttttttt taataaatgg
      961 tatttattaa tttatttgtt gtgtgtgttt ttttttttat aatatttttt tttttagcat
     1021 tgaattaaga agaaatcaaa ttgatgcggc cgcttagtta gcctccccca tctcccgatc
     1081 cggacgagtg ctggggcgtc ggtttccact atcggcgagt acttctacac agccatcggt
     1141 ccagacggcc gcgcttctgc gggcgatttg tgtacgcccg acagtcccgg ctccggatcg
     1201 gacgattgcg tcgcatcgac cctgcgccca agctgcatca tcgaaattgc cgtcaaccaa
     1261 gctctgatag agttggtcaa gaccaatgcg gagcatatac gcccggagcc gcggcgatcc
     1321 tgcaagctcc ggatgcctcc gctcgaagta gcgcgtctgc tgctccatac aagccaacca
     1381 cggcctccag aagaagatgt tggcgacctc gtattgggaa tccccgaaca tcgcctcgct
     1441 ccagtcaatg accgctgtta tgcggccatt gtccgtcagg acattgttgg agccgaaatc
     1501 cgcgtgcacg aggtgccgga cttcggggca gtcctcggcc caaagcatca gctcatcgag
     1561 agcctgcgcg acggacgcac tgacggtgtc gtccatcaca gtttgccagt gatacacatg
     1621 gggatcagca atcgcgcata tgaaatcacg ccatgtagtg tattgaccga ttccttgcgg
     1681 tccgaatggg ccgaacccgc tcgtctggct aagatcggcc gcagcgatcg catccatggc
     1741 ctccgcgacc ggctgcagaa cagcgggcag ttcggtttca ggcaggtctt gcaacgtgac
     1801 accctgtgca cggcgggaga tgcaataggt caggctctcg ctgaactccc caatgtcaag
     1861 cacttccgga atcgggagcg cggccgatgc aaagtgccga taaacataac gatctttgta
     1921 gaaaccatcg gcgcagctat ttacccgcag gacatatcca cgccctccta catcgaagct
     1981 gaaagcacga gattcttcgc cctccgagag ctgcatcagg tcggagacgc tgtcgaactt
     2041 ttcgatcaga aacttctcga cagacgtcgc ggtgagttca ggctttttca ttaggtcttg
     2101 ttgagaaatg ttaaattgat ccattttttg ctagctgtga aattagttta aaatacaaat
     2161 aaagagttat aataatatac agttgaataa aaaaaaaaaa aatgaattgg aaaatttatt
     2221 tttatatgaa gaaaaaaaaa ttttgaaaaa aaaaaaaaaa attaaaaaaa aaaaaaaaaa
     2281 aaaaaaaaat taaaaataat tcaactgtgg gtgacccaaa ttttatttaa aaaaaaaaaa
     2341 aaaatggatc acttttgggt ggttggaaaa aaaaaaaaaa aaaaattgaa aatataatgt
     2401 tagtcatatg attaatcatt aattaattct tttttttttt ttttttattt tttatttttt
     2461 ttttttcaat attccaatta tgacacacac taacatcaca aaatacaaaa ttttgggatc
     2521 cactactata acttcgtata atgtatgcta tacgaagtta tttaattaat taagatatca
     2581 agcttgatag ctctgcctac tgaaggccgg cagaggcggt ttgcgtattg ggcgctcttc
     2641 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2701 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2761 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2821 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2881 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2941 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3001 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3061 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3121 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3181 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3241 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3301 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3361 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3421 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3481 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3541 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     3601 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     3661 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     3721 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     3781 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     3841 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     3901 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     3961 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4021 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4081 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4141 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4201 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4261 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4321 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4381 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4441 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4501 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4561 gccacctgac gcgccctgta gcgggatca
//
