LOCUS       sequence-216060-        4610 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 4610)
  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 4610)
  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..4610
                     /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(1076..2113)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPDRE
                     MGEAN"
     protein_bind    2549..2582
                     /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(2668..2685)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2839..3427)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2919..2938)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3598..4389,4390..4458))
                     /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     4221..4240
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4459..4563)
                     /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 ttttaataaa tggtatttat taatttattt gttgtgtgtg tttttttttt
     1021 ataatatttt tttttttagc attgaattaa gaagaaatca aattgatgcg gccgcttagt
     1081 tagcctcccc catctcccga tccggacgag tgctggggcg tcggtttcca ctatcggcga
     1141 gtacttctac acagccatcg gtccagacgg ccgcgcttct gcgggcgatt tgtgtacgcc
     1201 cgacagtccc ggctccggat cggacgattg cgtcgcatcg accctgcgcc caagctgcat
     1261 catcgaaatt gccgtcaacc aagctctgat agagttggtc aagaccaatg cggagcatat
     1321 acgcccggag ccgcggcgat cctgcaagct ccggatgcct ccgctcgaag tagcgcgtct
     1381 gctgctccat acaagccaac cacggcctcc agaagaagat gttggcgacc tcgtattggg
     1441 aatccccgaa catcgcctcg ctccagtcaa tgaccgctgt tatgcggcca ttgtccgtca
     1501 ggacattgtt ggagccgaaa tccgcgtgca cgaggtgccg gacttcgggg cagtcctcgg
     1561 cccaaagcat cagctcatcg agagcctgcg cgacggacgc actgacggtg tcgtccatca
     1621 cagtttgcca gtgatacaca tggggatcag caatcgcgca tatgaaatca cgccatgtag
     1681 tgtattgacc gattccttgc ggtccgaatg ggccgaaccc gctcgtctgg ctaagatcgg
     1741 ccgcagcgat cgcatccatg gcctccgcga ccggctgcag aacagcgggc agttcggttt
     1801 caggcaggtc ttgcaacgtg acaccctgtg cacggcggga gatgcaatag gtcaggctct
     1861 cgctgaactc cccaatgtca agcacttccg gaatcgggag cgcggccgat gcaaagtgcc
     1921 gataaacata acgatctttg tagaaaccat cggcgcagct atttacccgc aggacatatc
     1981 cacgccctcc tacatcgaag ctgaaagcac gagattcttc gccctccgag agctgcatca
     2041 ggtcggagac gctgtcgaac ttttcgatca gaaacttctc gacagacgtc gcggtgagtt
     2101 caggcttttt cattaggtct tgttgagaaa tgttaaattg atccattttt tgctagctgt
     2161 gaaattagtt taaaatacaa ataaagagtt ataataatat acagttgaat aaaaaaaaaa
     2221 aaaatgaatt ggaaaattta tttttatatr aaaaaaaaaa aattttraaa aaaaaaaaaa
     2281 aaawwaaaaa aaaaaaaaaa aaaaaaaaaa atwaaaaawa attmaactgt gggtgaccca
     2341 aattttattt aaaaaaaaaa aaaaaatgga tcacttttgg gtggttggaa aaaaaaaaaa
     2401 aaaaaaattg aaaatataat gttagtcata tgattaatca ttaattaatt cttttttttt
     2461 ttttttttat tttttatttt tttttttcaa tattccaatt atgacacaca ctaacatcac
     2521 aaaatacaaa attttgggat ccactactat aacttcgtat aatgtatgct atacgaagtt
     2581 atttaattaa ttaagatatc aagcttgata gctctgccta ctgaaggccg gcagaggcgg
     2641 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2701 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2761 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2821 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2881 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2941 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3001 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3061 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3121 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3181 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3241 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3301 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3361 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3421 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3481 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3541 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3601 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3661 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3721 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3781 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3841 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3901 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3961 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4021 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4081 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4141 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4201 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4261 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4321 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4381 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4441 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4501 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4561 gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcgggatca
//
