LOCUS       sequence-234124-        6978 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses sphingosine kinase 1, which catalyzes the phosphorylation 
            of sphingosine to sphingosine-1-phosphate (S1P).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6978)
  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 6978)
  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: HsSPHK1
FEATURES             Location/Qualifiers
     source          1..6978
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(252..765)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(447..468)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     659..678
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        787..1228
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    787..904
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     1199..1222
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     1260..1279
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1260..1278
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1298..1321
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      2512..2759
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2528..2546)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2836..2853)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3007..3595)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3087..3106)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3766..4557,4558..4626))
                     /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     4389..4408
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(4722..5525)
                     /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(5526..5741)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      join(6342..6978,1..241)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 actacgtata ggaaatgttt acattttcgt attgttttcg attcactcta tgaatagttc
       61 ttactacaat ttttttgtct aaagagtaat actagagata aacataaaaa atgtagaggt
      121 cgagtttaga tgcaagttca aggagcgaaa ggtggatggg taggttatat agggatatag
      181 cacagagata tatagcaaag agatactttt gagcaatgtt tgtggaagcg gtattcgcaa
      241 tgggaagctc caccccggtt gataatcaga aaagccccaa aaacaggaag attgtataag
      301 caaatattta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
      361 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
      421 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
      481 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
      541 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
      601 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
      661 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
      721 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtgggga tcgatccact
      781 agtccacgga ttagaagccg ccgagcgggt gacagccctc cgaaggaaga ctctcctccg
      841 tgcgtcctcg tcttcaccgg tcgcgttcct gaaacgcaga tgtgcctcgc gccgcactgc
      901 tccgaacaat aaagattcta caatactagc ttttatggtt atgaagagga aaaattggca
      961 gtaacctggc cccacaaacc ttcaaatgaa cgaatcaaat taacaaccat aggatgataa
     1021 tgcgattagt tttttagcct tatttctggg gtaattaatc agcgaagcga tgatttttga
     1081 tctattaaca gatatataaa tgcaaaaact gcataaccac tttaactaat actttcaaca
     1141 ttttcggttt gtattacttc ttattcaaat gtaataaaag tatcaacaaa aaattgttaa
     1201 tatacctcta tactttaacg tcaaggagaa aaaaccccgg atcggactac tagcagctgt
     1261 aatacgactc actataggga atattaagct taccatggac tacaaggacg atgacgataa
     1321 gggatccatg gaccctgctg gtggtccaag gggtgttttg ccaagacctt gtagggtctt
     1381 ggtcttgttg aacccaaggg gtggtaaggg taaggcttta caattgttta gatctcatgt
     1441 ccaaccatta ttggctgaag cagaaatttc ttttactttg atgttgactg aaagaaggaa
     1501 ccacgctagg gaattggtca ggtctgaaga gttgggaagg tgggacgctt tggtcgttat
     1561 gtcaggtgac ggtttgatgc acgaagtcgt caacggttta atggagaggc cagactggga
     1621 gactgctatt cagaaacctt tgtgttcttt gccagcaggt tctggtaacg cattggcagc
     1681 ttctttgaac cactacgctg gatacgagca ggtcactaat gaagacttgt tgactaattg
     1741 tactttattg ttgtgtagaa gattgttgtc tccaatgaac ttgttgtcat tgcatacagc
     1801 atctggttta agattgtttt ctgtcttatc attggcttgg ggttttattg ctgatgttga
     1861 tttggaatct gaaaagtata gaagattggg tgaaatgaga ttcactttgg gtactttttt
     1921 gagattagca gctttgagaa cttatagagg taggttggct tatttgcctg ttggtagggt
     1981 cggttcaaag acaccagctt ctccagttgt cgtccagcag ggtccagttg atgctcactt
     2041 ggtccctttg gaggaaccag ttccatctca ctggacagtc gttccagacg aggacttcgt
     2101 cttggttttg gctttgttac attctcattt gggatctgag atgttcgctg ctccaatggg
     2161 tagatgcgct gctggtgtca tgcatttgtt ctacgtcagg gctggtgtct caagggctat
     2221 gttgttgaga ttatttttgg ctatggaaaa aggtagacat atggaatacg agtgcccata
     2281 cttggtttac gtcccagttg tcgctttcag gttggagcca aaggatggta agggtgtctt
     2341 cgcagtcgat ggtgagttga tggtctctga agctgtccaa ggtcaagtcc acccaaacta
     2401 cttctggatg gtctcaggtt gcgttgagcc accaccatct tggaagccac aacaaatgcc
     2461 accacctgag gaacctttat aatgactcga gcatgcatct agagggccgc atcatgtaat
     2521 tagttatgtc acgcttacat tcacgccctc cccccacatc cgctctaacc gaaaaggaag
     2581 gagttagaca acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa
     2641 gaacgttatt tatatttcaa atttttcttt tttttctgta cagacgcgtg tacgcatgta
     2701 acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct ttaatttgca
     2761 agctgcggcc ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2821 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2881 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2941 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3001 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3061 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3121 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3181 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3241 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3301 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3361 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3421 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3481 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3541 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3601 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3661 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3721 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3781 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3841 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3901 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3961 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4021 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4081 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4141 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4201 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4261 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4321 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4381 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4441 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4501 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4561 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4621 actcatactc ttcctttttc aatgggtaat aactgatata attaaattga agctctaatt
     4681 tgtgagttta gtatacatgc atttacttat aatacagttt tttagttttg ctggccgcat
     4741 cttctcaaat atgcttccca gcctgctttt ctgtaacgtt caccctctac cttagcatcc
     4801 cttccctttg caaatagtcc tcttccaaca ataataatgt cagatcctgt agagaccaca
     4861 tcatccacgg ttctatactg ttgacccaat gcgtctccct tgtcatctaa acccacaccg
     4921 ggtgtcataa tcaaccaatc gtaaccttca tctcttccac ccatgtctct ttgagcaata
     4981 aagccgataa caaaatcttt gtcgctcttc gcaatgtcaa cagtaccctt agtatattct
     5041 ccagtagata gggagccctt gcatgacaat tctgctaaca tcaaaaggcc tctaggttcc
     5101 tttgttactt cttctgccgc ctgcttcaaa ccgctaacaa tacctgggcc caccacaccg
     5161 tgtgcattcg taatgtctgc ccattctgct attctgtata cacccgcaga gtactgcaat
     5221 ttgactgtat taccaatgtc agcaaatttt ctgtcttcga agagtaaaaa attgtacttg
     5281 gcggataatg cctttagcgg cttaactgtg ccctccatgg aaaaatcagt caagatatcc
     5341 acatgtgttt ttagtaaaca aattttggga cctaatgctt caactaactc cagtaattcc
     5401 ttggtggtac gaacatccaa tgaagcacac aagtttgttt gcttttcgtg catgatatta
     5461 aatagcttgg cagcaacagg actaggatga gtagcagcac gttccttata tgtagctttc
     5521 gacatgattt atcttcgttt cctgcaggtt tttgttctgt gcagttgggt taagaatact
     5581 gggcaatttc atgtttcttc aacactacat atgcgtatat ataccaatct aagtctgtgc
     5641 tccttccttc gttcttcctt ctgttcggag attaccgaat caaaaaaatt tcaaggaaac
     5701 cgaaatcaaa aaaaagaata aaaaaaaaat gatgaattga aaagctagct tatcgatgat
     5761 aagctgtcaa acatgagaat taattccacg gactatagac tatacctagt atactccgtc
     5821 tactgtacga tacacttccg ctcaggtcct tgtcctttaa cgaggcctta ccactctttt
     5881 gttactctat tgatccagct cagcaaaggc agtgtgatct aagattctat cttcgcgatg
     5941 tagtaaaact agctagaccg agaaagagac tagaaatgca aaaggcactt ctacaatggc
     6001 tgccatcatt attatccgat gtgacgctgc agcttctcaa tgatattcga atacgctttg
     6061 aggagataca gcctaatatc cgacaaactg ttttacagat ttacgatcgt acttgttacc
     6121 catcattgaa ttttgaacat ccgaacctgg gagttttccc tgaaacagat agtatatttg
     6181 aacctgtata ataatatata gtctagcgct ttacggaaga caatgtatgt atttcggttc
     6241 ctggagaaac tattgcatct attgcatagg taatcttgca cgtcgcatcc ccggttcatt
     6301 ttctgcgttt ccatcttgca cttcaatagc atatctttgt taacgaagca tctgtgcttc
     6361 attttgtaga acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct
     6421 gcatttttac agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg
     6481 cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct
     6541 gagctgcatt tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat
     6601 ctatacttct tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc
     6661 atcttagatt actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt
     6721 tgcactgtag gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat
     6781 aaaaaaagcc tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt
     6841 ttttcaagat aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg
     6901 tgaacagaaa gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct
     6961 tctattttgt ctctatat
//
