LOCUS       sequence-448436-        4546 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid donor template for CRISPR/Cas9 mediated UGCG knockout by 
            tdTomato insertion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4546)
  AUTHORS   Sato Y, Osada E, Ushiki T, Maeda T, Manome Y
  TITLE     UDP-glucose ceramide glucosyltransferase specifically upregulated in
            plasmacytoid dendritic cells regulates type I interferon production 
            upon CpG stimulation.
  JOURNAL   Biochem Biophys Res Commun. 2024 Sep 18;733:150703. doi: 
            10.1016/j.bbrc.2024.150703.
  PUBMED    39307111
REFERENCE   2  (bases 1 to 4546)
  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: "Biochem 
            Biophys Res Commun."; date: "2024-09-18"; volume: "733:150703. doi";
            pages: " 10.1016/j.bbrc.2024.150703"
COMMENT     Sequence Label: pUCmini-UGCG-tdTomato
FEATURES             Location/Qualifiers
     source          1..4546
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        339..849
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     768..786
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     regulatory      865..874
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             871..2301
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     polyA_signal    2307..2418
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2791..3379)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2871..2890)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3550..4341,4342..4410))
                     /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     4173..4192
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4411..4515)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccaatgatct cgagtcacat ataaaataca ctaacaatag ctgatgagct ttaaaaaatc
       61 gcaaaaaatc tcataatgtt ttaagaaagt ttatgaattt gtgttggacc gcattcaaag
      121 ccatgcttac ctggttctac ttcccttcat gtttagatcc tcttttagaa agatgtcaag
      181 ttttaaaaat cttaacttag aatttaggga tttattgata gcttgtcagt gctttgattt
      241 gacactaatg tattctatag aaataatttt tatatcattt gctttttagc cgattacacg
      301 tcaacgcatg ctggggatgc ggtgggctga attcccacgg ggttggggtt gcgccttttc
      361 caaggcagcc ctgggtttgc gcagggacgc ggctgctctg ggcgtggttc cgggaaacgc
      421 agcggcgccg accctgggtc tcgcacattc ttcacgtccg ttcgcagcgt cacccggatc
      481 ttcgccgcta cccttgtggg ccccccggcg acgcttcctg ctccgcccct aagtcgggaa
      541 ggttccttgc ggttcgcggc gtgccggacg tgacaaacgg aagccgcacg tctcactagt
      601 accctcgcag acggacagcg ccagggagca atggcagcgc gccgaccgcg atgggctgtg
      661 gccaatagcg gctgctcagc ggggcgcgcc gagagcagcg gccgggaagg ggcggtgcgg
      721 gaggcggggt gtggggcggt agtgtgggcc ctgttcctgc ccgcgcggtg ttccgcattc
      781 tgcaagcctc cggagcgcac gtcggcagtc ggctccctcg ttgaccgaat caccgacctc
      841 tctccccagg gggatccacc ggtcgccacc atggtgagca agggcgagga ggtcatcaaa
      901 gagttcatgc gcttcaaggt gcgcatggag ggctccatga acggccacga gttcgagatc
      961 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
     1021 aagggcggcc ccctgccctt cgcctgggac atcctgtccc cccagttcat gtacggctcc
     1081 aaggcgtacg tgaagcaccc cgccgacatc cccgattaca agaagctgtc cttccccgag
     1141 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gtctggtgac cgtgacccag
     1201 gactcctccc tgcaggacgg cacgctgatc tacaaggtga agatgcgcgg caccaacttc
     1261 ccccccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc caccgagcgc
     1321 ctgtaccccc gcgacggcgt gctgaagggc gagatccacc aggccctgaa gctgaaggac
     1381 ggcggccact acctggtgga gttcaagacc atctacatgg ccaagaagcc cgtgcaactg
     1441 cccggctact actacgtgga caccaagctg gacatcacct cccacaacga ggactacacc
     1501 atcgtggaac agtacgagcg ctccgagggc cgccaccacc tgttcctggg gcatggcacc
     1561 ggcagcaccg gcagcggcag ctccggcacc gcctcctccg aggacaacaa catggccgtc
     1621 atcaaagagt tcatgcgctt caaggtgcgc atggagggct ccatgaacgg ccacgagttc
     1681 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
     1741 gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccccca gttcatgtac
     1801 ggctccaagg cgtacgtgaa gcaccccgcc gacatccccg attacaagaa gctgtccttc
     1861 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggtct ggtgaccgtg
     1921 acccaggact cctccctgca ggacggcacg ctgatctaca aggtgaagat gcgcggcacc
     1981 aacttccccc ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctccacc
     2041 gagcgcctgt acccccgcga cggcgtgctg aagggcgaga tccaccaggc cctgaagctg
     2101 aaggacggcg gccactacct ggtggagttc aagaccatct acatggccaa gaagcccgtg
     2161 caactgcccg gctactacta cgtggacacc aagctggaca tcacctccca caacgaggac
     2221 tacaccatcg tggaacagta cgagcgctcc gagggccgcc accacctgtt cctgtacggc
     2281 atggacgagc tgtacaagta ggtcgactgt gccttctagt tgccagccat ctgttgtttg
     2341 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     2401 aaatgaggaa attgcatcat cgataagaag ccaactgaca aacagcctta tagcaagctc
     2461 ccaggtgtct ctcttctgaa accactgaaa ggggtagatc ctaacttaat caacaacctg
     2521 gaaacattct ttgaattgga ttatcccaaa gtaagttcag tgttacggtt ttttgttttg
     2581 ttttgttttg ttttgttccc ctcatgataa acatttaggg attttaacac tgtataaggt
     2641 gaaggtatta aaaattcatg atgatattgg cctcataaaa tataaccttt agtagtagtc
     2701 atcacagaac agagctagca tcggaaagaa catgtgagca aaaggccagc aaaaggccag
     2761 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2821 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2881 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2941 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3001 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3061 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3121 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3181 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3241 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3301 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3361 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3421 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3481 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3541 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3601 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3661 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3721 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3781 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3841 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     3901 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     3961 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4021 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4081 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4141 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4201 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4261 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4321 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4381 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4441 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4501 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
