LOCUS       sequence-234128-        7434 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ceramide kinase, which catalyzes the phosphorylation of 
            ceramide to form ceramide 1-phosphate.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7434)
  AUTHORS   Kharel Y, Agah S, Huang T, Mendelson AJ, Eletu OT, Barkey-Bircann P,
            Gesualdi J, Smith JS, Santos WL, Lynch KR
  TITLE     Saccharomyces cerevisiae as a platform for assessing sphingolipid 
            lipid kinase inhibitors.
  JOURNAL   PLoS One. 2018 Apr 19;13(4):e0192179. doi: 
            10.1371/journal.pone.0192179. eCollection 2018.
  PUBMED    29672528
REFERENCE   2  (bases 1 to 7434)
  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: "2018-04-19"; volume: "13(4):e0192179. doi"; pages: " 
            10.1371/journal.pone.0192179. eCollection 2018"
COMMENT     Sequence Label: HsCERK
FEATURES             Location/Qualifiers
     source          1..7434
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        26..467
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    26..143
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     438..461
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     499..518
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        499..517
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             537..560
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      2207..2454
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2223..2241)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2531..2548)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2702..3290)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2782..2801)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3461..4252,4253..4321))
                     /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     4084..4103
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(4417..5220)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(5221..5436)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      6037..6914
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(join(6925..7434,1..4))
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7120..7141)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7332..7351
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcgtggggat cgatccacta gtccacggat tagaagccgc cgagcgggtg acagccctcc
       61 gaaggaagac tctcctccgt gcgtcctcgt cttcaccggt cgcgttcctg aaacgcagat
      121 gtgcctcgcg ccgcactgct ccgaacaata aagattctac aatactagct tttatggtta
      181 tgaagaggaa aaattggcag taacctggcc ccacaaacct tcaaatgaac gaatcaaatt
      241 aacaaccata ggatgataat gcgattagtt ttttagcctt atttctgggg taattaatca
      301 gcgaagcgat gatttttgat ctattaacag atatataaat gcaaaaactg cataaccact
      361 ttaactaata ctttcaacat tttcggtttg tattacttct tattcaaatg taataaaagt
      421 atcaacaaaa aattgttaat atacctctat actttaacgt caaggagaaa aaaccccgga
      481 tcggactact agcagctgta atacgactca ctatagggaa tattaagctt accatggact
      541 acaaggacga tgacgataag ggatccatgg gtgctacagg tgctgctgag ccattgcagt
      601 ctgtcttgtg ggtcaaacag cagaggtgcg ctgtttcttt ggagcctgca agagctttat
      661 tgaggtggtg gaggtcacca ggtccaggtg caggtgcacc tggtgcagat gcttgttcag
      721 ttccagtctc tgagattatt gctgttgaag agactgatgt ccacggtaag catcaaggat
      781 ctggtaagtg gcagaagatg gagaagcctt acgcattcac tgtccattgt gttaagaggg
      841 ctagaaggca caggtggaag tgggctcagg tcactttttg gtgtccagag gagcagttgt
      901 gccatttgtg gttgcaaact ttgagagaaa tgttggaaaa gttgacttca aggccaaagc
      961 atttgttggt ttttattaat ccattcggtg gtaagggtca aggtaaaaga atatatgaga
     1021 gaaaggtcgc tcctttgttc acattggcat caataacaac tgatattatt gttacagaac
     1081 acgctaacca ggctaaagaa actttgtacg aaattaatat tgataagtac gatggaatag
     1141 tctgcgtcgg tggtgatggt atgttctcag aagttttgca tggtttgatt ggtagaactc
     1201 agaggtcagc aggtgtcgac caaaaccacc ctagagctgt cttggttcct tcttctttga
     1261 gaattggtat aatacctgct ggttctactg actgtgtctg ctactctaca gtcggtactt
     1321 cagacgcaga aacttctgct ttacacattg ttgtcggtga ttctttagca atggatgttt
     1381 cttcagttca tcataattct acattgttga gatactcagt ttcattgttg ggttatggtt
     1441 tctatggaga tattattaag gactcagaaa agaagagatg gttgggattg gctaggtacg
     1501 acttctcagg tttgaaaaca tttttgtctc atcactgcta cgagggtaca gtctcattct
     1561 tgccagctca acacactgtc ggttcaccaa gagacagaaa gccatgcagg gcaggttgtt
     1621 ttgtttgtag acaatctaaa caacaattag aagaagaaca gaaaaaagca ttgtacggtt
     1681 tggaagctgc agaggacgtc gaagaatggc aagttgtctg cggtaaattt ttggctatta
     1741 atgctactaa tatgtcttgt gcttgcagga gatcaccaag gggtttatct ccagctgctc
     1801 acttgggtga cggttcttct gatttgatat tgattagaaa atgttctaga ttcaattttt
     1861 tgagattctt aattagacat actaatcaac aagatcaatt tgatttcaca tttgttgagg
     1921 tttacagagt taaaaagttt cagtttactt ctaaacatat ggaggatgaa gattctgatt
     1981 taaaagaagg tggtaagaag aggtttggtc atatttgctc ttctcatcca tcttgttgct
     2041 gtacagtttc taattcttca tggaattgcg acggtgaggt tttgcactct ccagcaattg
     2101 aggttagggt ccactgtcag ttggtcaggt tgttcgcaag gggtattgag gaaaacccaa
     2161 aacctgattc tcactcttaa ctcgagcatg catctagagg gccgcatcat gtaattagtt
     2221 atgtcacgct tacattcacg ccctcccccc acatccgctc taaccgaaaa ggaaggagtt
     2281 agacaacctg aagtctaggt ccctatttat ttttttatag ttatgttagt attaagaacg
     2341 ttatttatat ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc atgtaacatt
     2401 atactgaaaa ccttgcttga gaaggttttg ggacgctcga aggctttaat ttgcaagctg
     2461 cggccctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2521 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2581 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2641 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2701 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2761 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2821 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2881 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2941 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3001 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3061 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3121 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3181 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3241 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3301 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3361 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3421 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3481 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3541 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3601 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3661 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     3721 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     3781 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     3841 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     3901 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     3961 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4021 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4081 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4141 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4201 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4261 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4321 tactcttcct ttttcaatgg gtaataactg atataattaa attgaagctc taatttgtga
     4381 gtttagtata catgcattta cttataatac agttttttag ttttgctggc cgcatcttct
     4441 caaatatgct tcccagcctg cttttctgta acgttcaccc tctaccttag catcccttcc
     4501 ctttgcaaat agtcctcttc caacaataat aatgtcagat cctgtagaga ccacatcatc
     4561 cacggttcta tactgttgac ccaatgcgtc tcccttgtca tctaaaccca caccgggtgt
     4621 cataatcaac caatcgtaac cttcatctct tccacccatg tctctttgag caataaagcc
     4681 gataacaaaa tctttgtcgc tcttcgcaat gtcaacagta cccttagtat attctccagt
     4741 agatagggag cccttgcatg acaattctgc taacatcaaa aggcctctag gttcctttgt
     4801 tacttcttct gccgcctgct tcaaaccgct aacaatacct gggcccacca caccgtgtgc
     4861 attcgtaatg tctgcccatt ctgctattct gtatacaccc gcagagtact gcaatttgac
     4921 tgtattacca atgtcagcaa attttctgtc ttcgaagagt aaaaaattgt acttggcgga
     4981 taatgccttt agcggcttaa ctgtgccctc catggaaaaa tcagtcaaga tatccacatg
     5041 tgtttttagt aaacaaattt tgggacctaa tgcttcaact aactccagta attccttggt
     5101 ggtacgaaca tccaatgaag cacacaagtt tgtttgcttt tcgtgcatga tattaaatag
     5161 cttggcagca acaggactag gatgagtagc agcacgttcc ttatatgtag ctttcgacat
     5221 gatttatctt cgtttcctgc aggtttttgt tctgtgcagt tgggttaaga atactgggca
     5281 atttcatgtt tcttcaacac tacatatgcg tatatatacc aatctaagtc tgtgctcctt
     5341 ccttcgttct tccttctgtt cggagattac cgaatcaaaa aaatttcaag gaaaccgaaa
     5401 tcaaaaaaaa gaataaaaaa aaaatgatga attgaaaagc tagcttatcg atgataagct
     5461 gtcaaacatg agaattaatt ccacggacta tagactatac ctagtatact ccgtctactg
     5521 tacgatacac ttccgctcag gtccttgtcc tttaacgagg ccttaccact cttttgttac
     5581 tctattgatc cagctcagca aaggcagtgt gatctaagat tctatcttcg cgatgtagta
     5641 aaactagcta gaccgagaaa gagactagaa atgcaaaagg cacttctaca atggctgcca
     5701 tcattattat ccgatgtgac gctgcagctt ctcaatgata ttcgaatacg ctttgaggag
     5761 atacagccta atatccgaca aactgtttta cagatttacg atcgtacttg ttacccatca
     5821 ttgaattttg aacatccgaa cctgggagtt ttccctgaaa cagatagtat atttgaacct
     5881 gtataataat atatagtcta gcgctttacg gaagacaatg tatgtatttc ggttcctgga
     5941 gaaactattg catctattgc ataggtaatc ttgcacgtcg catccccggt tcattttctg
     6001 cgtttccatc ttgcacttca atagcatatc tttgttaacg aagcatctgt gcttcatttt
     6061 gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     6121 tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca
     6181 tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct
     6241 gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa agaatctata
     6301 cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac aaagcatctt
     6361 agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa ctttttgcac
     6421 tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct tccataaaaa
     6481 aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt gcattttttc
     6541 aagataaagg catccccgat tatattctat accgatgtgg attgcgcata ctttgtgaac
     6601 agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg tttcttctat
     6661 tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt tcgattcact
     6721 ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag ataaacataa
     6781 aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat gggtaggtta
     6841 tatagggata tagcacagag atatatagca aagagatact tttgagcaat gtttgtggaa
     6901 gcggtattcg caatgggaag ctccaccccg gttgataatc agaaaagccc caaaaacagg
     6961 aagattgtat aagcaaatat ttaaattgta agcgttaata ttttgttaaa attcgcgtta
     7021 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     7081 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     7141 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     7201 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     7261 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     7321 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     7381 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggc
//
