LOCUS       sequence-205374-        4613 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  generic knockin vector for C-terminal tagging with GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4613)
  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 4613)
  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: pDM1355
FEATURES             Location/Qualifiers
     source          1..4613
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    769..802
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1078..2115)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPDRE
                     MGEAN"
     protein_bind    2552..2585
                     /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(2671..2688)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2842..3430)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2922..2941)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3601..4392,4393..4461))
                     /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     4224..4243
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4462..4566)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 actagtggtg gttcaggagg tagttcaaaa ggtgaagaat tatttacagg tgttgttcca
       61 attttagttg aattagatgg tgatgttaat ggtcataaat tttcagtttc aggtgaaggt
      121 gaaggtgatg caacatatgg taaattaaca ttaaaattta tttgtacaac aggtaaatta
      181 ccagttccat ggccaacatt agttacaaca tttacatatg gtgttcaatg tttttcaaga
      241 tatccagatc atatgaaaca acatgatttt tttaaatcag caatgccaga aggttatgtt
      301 caagaaagaa caattttttt taaagatgat ggtaattata aaacaagagc agaagttaaa
      361 tttgaaggtg atacattagt taatagaatt gaattaaaag gtattgattt taaagaagat
      421 ggtaatattt taggtcataa attagaatat aattataatt cacataatgt ttatattatg
      481 gcagataaac aaaaaaatgg tattaaagtt aattttaaaa ttagacataa tattgaagat
      541 ggttcagttc aattagcaga tcattatcaa caaaatacac caattggtga tggtccagtt
      601 ttattaccag ataatcatta tttatcaaca caatcagcat tatcaaaaga tccaaatgaa
      661 aaaagagatc atatggtttt attagaattt gttacagcag caggtattac acatggtatg
      721 gatgaattat ataaataaat aaaataaata aattgtagat ctgatatcat aacttcgtat
      781 aatgtatgct atacgaagtt atactactct cgagatttta tttaatatac taaataataa
      841 aaaagttaaa aaatgatcat tggataaatt ttttataatt ataaataaag ataataattt
      901 tttttttaac aaaactaaaa ataaaaataa taaaataatt gttaaaatag gttttttttt
      961 tttttttttt tttttaataa atggtattta ttaatttatt tgttgtgtgt gttttttttt
     1021 ttataatatt ttttttttta gcattgaatt aagaagaaat caaattgatg cggccgctta
     1081 gttagcctcc cccatctccc gatccggacg agtgctgggg cgtcggtttc cactatcggc
     1141 gagtacttct acacagccat cggtccagac ggccgcgctt ctgcgggcga tttgtgtacg
     1201 cccgacagtc ccggctccgg atcggacgat tgcgtcgcat cgaccctgcg cccaagctgc
     1261 atcatcgaaa ttgccgtcaa ccaagctctg atagagttgg tcaagaccaa tgcggagcat
     1321 atacgcccgg agccgcggcg atcctgcaag ctccggatgc ctccgctcga agtagcgcgt
     1381 ctgctgctcc atacaagcca accacggcct ccagaagaag atgttggcga cctcgtattg
     1441 ggaatccccg aacatcgcct cgctccagtc aatgaccgct gttatgcggc cattgtccgt
     1501 caggacattg ttggagccga aatccgcgtg cacgaggtgc cggacttcgg ggcagtcctc
     1561 ggcccaaagc atcagctcat cgagagcctg cgcgacggac gcactgacgg tgtcgtccat
     1621 cacagtttgc cagtgataca catggggatc agcaatcgcg catatgaaat cacgccatgt
     1681 agtgtattga ccgattcctt gcggtccgaa tgggccgaac ccgctcgtct ggctaagatc
     1741 ggccgcagcg atcgcatcca tggcctccgc gaccggctgc agaacagcgg gcagttcggt
     1801 ttcaggcagg tcttgcaacg tgacaccctg tgcacggcgg gagatgcaat aggtcaggct
     1861 ctcgctgaac tccccaatgt caagcacttc cggaatcggg agcgcggccg atgcaaagtg
     1921 ccgataaaca taacgatctt tgtagaaacc atcggcgcag ctatttaccc gcaggacata
     1981 tccacgccct cctacatcga agctgaaagc acgagattct tcgccctccg agagctgcat
     2041 caggtcggag acgctgtcga acttttcgat cagaaacttc tcgacagacg tcgcggtgag
     2101 ttcaggcttt ttcattaggt cttgttgaga aatgttaaat tgatccattt tttgctagct
     2161 gtgaaattag tttaaaatac aaataaagag ttataataat atacagttga ataaaaaaaa
     2221 aaaaaatgaa ttggaaaatt tatttttata tgaagaaaaa aaaattttga aaaaaaaaaa
     2281 aaaaattaaa aaaaaaaaaa aaaaaaaaaa aaattaaaaa taattcaact gtgggtgacc
     2341 caaattttat ttaaaaaaaa aaaaaaaatg gatcactttt gggtggttgg aaaaaaaaaa
     2401 aaaaaaaaat tgaaaatata atgttagtca tatgattaat cattaattaa ttcttttttt
     2461 tttttttttt attttttatt tttttttttt caatattcca attatgacac acactaacat
     2521 cacaaaatac aaaattttgg gatccactac tataacttcg tataatgtat gctatacgaa
     2581 gttatttaat taattaagat atcaagcttg atagctctgc ctactgaagg ccggcagagg
     2641 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2701 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2761 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2821 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2881 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2941 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3001 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3061 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3121 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3181 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3241 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     3301 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3361 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3421 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3481 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3541 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3601 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3661 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3721 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3781 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3841 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3901 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3961 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4021 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4081 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4141 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4201 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4261 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4321 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4381 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4441 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4501 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4561 tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggga tca
//
