LOCUS       sequence-232881-        4584 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 4584)
  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 4584)
  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..4584
                     /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(1050..2087)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPDRE
                     MGEAN"
     protein_bind    2523..2556
                     /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(2642..2659)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2813..3401)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2893..2912)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3572..4363,4364..4432))
                     /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     4195..4214
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4433..4537)
                     /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 ttttttttaa taaatggtat
      961 ttattaattt atttgttgtg tgtgtttttt ttttataata tttttttttt tagcattgaa
     1021 ttaagaagaa atcaaattga tgcggccgct tagttagcct cccccatctc ccgatccgga
     1081 cgagtgctgg ggcgtcggtt tccactatcg gcgagtactt ctacacagcc atcggtccag
     1141 acggccgcgc ttctgcgggc gatttgtgta cgcccgacag tcccggctcc ggatcggacg
     1201 attgcgtcgc atcgaccctg cgcccaagct gcatcatcga aattgccgtc aaccaagctc
     1261 tgatagagtt ggtcaagacc aatgcggagc atatacgccc ggagccgcgg cgatcctgca
     1321 agctccggat gcctccgctc gaagtagcgc gtctgctgct ccatacaagc caaccacggc
     1381 ctccagaaga agatgttggc gacctcgtat tgggaatccc cgaacatcgc ctcgctccag
     1441 tcaatgaccg ctgttatgcg gccattgtcc gtcaggacat tgttggagcc gaaatccgcg
     1501 tgcacgaggt gccggacttc ggggcagtcc tcggcccaaa gcatcagctc atcgagagcc
     1561 tgcgcgacgg acgcactgac ggtgtcgtcc atcacagttt gccagtgata cacatgggga
     1621 tcagcaatcg cgcatatgaa atcacgccat gtagtgtatt gaccgattcc ttgcggtccg
     1681 aatgggccga acccgctcgt ctggctaaga tcggccgcag cgatcgcatc catggcctcc
     1741 gcgaccggct gcagaacagc gggcagttcg gtttcaggca ggtcttgcaa cgtgacaccc
     1801 tgtgcacggc gggagatgca ataggtcagg ctctcgctga actccccaat gtcaagcact
     1861 tccggaatcg ggagcgcggc cgatgcaaag tgccgataaa cataacgatc tttgtagaaa
     1921 ccatcggcgc agctatttac ccgcaggaca tatccacgcc ctcctacatc gaagctgaaa
     1981 gcacgagatt cttcgccctc cgagagctgc atcaggtcgg agacgctgtc gaacttttcg
     2041 atcagaaact tctcgacaga cgtcgcggtg agttcaggct ttttcattag gtcttgttga
     2101 gaaatgttaa attgatccat tttttgctag ctgtgaaatt agtttaaaat acaaataaag
     2161 agttataata atatacagtt gaataaaaaa aaaaaaaatr aattggaaaa tttattttta
     2221 tatraaaaaa aaaaaatttt raaaaaaaaa aaaaaaawta aaaaaaaaaa aaaaaaaaaa
     2281 aaaaattaaa aawaattcaa ctgtgggtga cccaaatttt atttaaaaaa aaaaaaaaaa
     2341 tggatcactt ttgggtggtt ggaaaaaaaa aaaaaaaaaa attgaaaata taatgttagt
     2401 catatgatta atcattaatt aattcttttt tttttttttt ttatttttta tttttttttt
     2461 tcaatattcc aattatgaca cacactaaca tcacaaaata caaaattttg ggatccacta
     2521 ctataacttc gtataatgta tgctatacga agttatttaa ttaattaaga tatcaagctt
     2581 gatagctctg cctactgaag gccggcagag gcggtttgcg tattgggcgc tcttccgctt
     2641 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2701 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2761 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2821 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2881 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2941 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3001 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3061 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3121 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3181 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3241 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3301 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3361 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3421 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3481 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3541 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3601 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3661 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3721 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3781 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3841 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3901 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3961 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4021 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4081 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4141 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4201 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4261 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4321 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4381 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4441 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4501 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4561 ctgacgcgcc ctgtagcggg atca
//
