LOCUS       sequence-234125-        7712 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses sphingosine kinase 2, 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 7712)
  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 7712)
  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: HsSPHK2
FEATURES             Location/Qualifiers
     source          1..7712
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     264..287
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     325..344
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        325..343
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             363..386
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      2311..2558
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2327..2345)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2635..2652)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2806..3394)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2886..2905)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3565..4356,4357..4425))
                     /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     4188..4207
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(4521..5324)
                     /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(5325..5540)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      6141..7018
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(7029..7542)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7224..7245)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7436..7455
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(7564..7712,1..293)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    7564..7681
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
ORIGIN
        1 tggttatgaa gaggaaaaat tggcagtaac ctggccccac aaaccttcaa atgaacgaat
       61 caaattaaca accataggat gataatgcga ttagtttttt agccttattt ctggggtaat
      121 taatcagcga agcgatgatt tttgatctat taacagatat ataaatgcaa aaactgcata
      181 accactttaa ctaatacttt caacattttc ggtttgtatt acttcttatt caaatgtaat
      241 aaaagtatca acaaaaaatt gttaatatac ctctatactt taacgtcaag gagaaaaaac
      301 cccggatcgg actactagca gctgtaatac gactcactat agggaatatt aagcttacca
      361 tggactacaa ggacgatgac gataagggat ccatggcccc gcccccaccg ccactggctg
      421 ccagcacccc gctcctccat ggcgagtttg gctcctaccc agcccgaggc ccacgctttg
      481 ccctcaccct tacatcgcag gccctgcaca tacagcggct gcgccccaaa cctgaagcca
      541 ggccccgggg tggcctggtc ccgttggccg aggtctcagg ctgctgcacc ctgcgaagcc
      601 gcagcccctc agactcagcg gcctacttct gcatctacac ctaccctcgg ggccggcgcg
      661 gggcccggcg cagagccact cgcaccttcc gggcagatgg ggccgccacc tacgaagaga
      721 accgtgccga ggcccagcgc tgggccactg ccctcacctg tctgctccga ggactgccac
      781 tgcccgggga tggggagatc acccctgacc tgctacctcg gccgccccgg ttgcttctat
      841 tggtcaatcc ctttgggggt cggggcctgg cctggcagtg gtgtaagaac cacgtgcttc
      901 ccatgatctc tgaagctggg ctgtccttca acctcatcca gacagaacga cagaaccacg
      961 cccgggagct ggtccagggg ctgagcctga gtgagtggga tggcatcgtc acggtctcgg
     1021 gagacgggct gctccatgag gtgctgaacg ggctcctaga tcgccctgac tgggaggaag
     1081 ctgtgaagat gcctgtgggc atcctcccct gcggctcggg caacgcgctg gccggagcag
     1141 tgaaccagca cgggggattt gagccagccc tgggcctcga cctgttgctc aactgctcac
     1201 tgttgctgtg ccggggtggt ggccacccac tggacctgct ctccgtgacg ctggcctcgg
     1261 gctcccgctg tttctccttc ctgtctgtgg cctggggctt cgtgtcagat gtggatatcc
     1321 agagcgagcg cttcagggcc ttgggcagtg cccgcttcac actgggcacg gtgctgggcc
     1381 tcgccacact gcacacctac cgcggacgcc tctcctacct ccccgccact gtggaacctg
     1441 cctcgcccac ccctgcccat agcctgcctc gtgccaagtc ggagctgacc ctaaccccag
     1501 acccagcccc gcccatggcc cactcacccc tgcatcgttc tgtgtctgac ctgcctcttc
     1561 ccctgcccca gcctgccctg gcctctcctg gctcgccaga acccctgccc atcctgtccc
     1621 tcaacggtgg gggcccagag ctggctgggg actggggtgg ggctggggat gctccgctgt
     1681 ccccggaccc actgctgtct tcacctcctg gctctcccaa ggcagctcta cactcacccg
     1741 tctccgaagg ggcccccgta attcccccat cctctgggct cccacttccc acccctgatg
     1801 cccgggtagg ggcctccacc tgcggcccgc ccgaccacct gctgcctccg ctgggcaccc
     1861 cgctgccccc agactgggtg acgctggagg gggactttgt gctcatgttg gccatctcgc
     1921 ccagccacct aggcgctgac ctggtggcag ctccgcatgc gcgcttcgac gacggcctgg
     1981 tgcacctgtg ctgggtgcgt agcggcatct cgcgggctgc gctgctgcgc cttttcttgg
     2041 ccatggagcg tggtagccac ttcagcctgg gctgtccgca gctgggctac gccgcggccc
     2101 gtgccttccg cctagagccg ctcacaccac gcggcgtgct cacagtggac ggggagcagg
     2161 tggagtatgg gccgctacag gcacagatgc accctggcat cggtacactg ctcactgggc
     2221 ctcctggctg cccggggcgg gagcccaagg gcaattctgc agatatccag cacagtggcg
     2281 gccgctcgag catgcatcta gagggccgca tcatgtaatt agttatgtca cgcttacatt
     2341 cacgccctcc ccccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct
     2401 aggtccctat ttattttttt atagttatgt tagtattaag aacgttattt atatttcaaa
     2461 tttttctttt ttttctgtac agacgcgtgt acgcatgtaa cattatactg aaaaccttgc
     2521 ttgagaaggt tttgggacgc tcgaaggctt taatttgcaa gctgcggccc tgcattaatg
     2581 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     2641 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2701 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2761 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2821 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2881 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2941 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3001 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3061 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3121 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3181 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3241 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3301 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3361 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3421 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3481 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3541 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3601 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3661 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3721 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3781 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3841 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     3901 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3961 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4021 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4081 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4141 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4201 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4261 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4321 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4381 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4441 atgggtaata actgatataa ttaaattgaa gctctaattt gtgagtttag tatacatgca
     4501 tttacttata atacagtttt ttagttttgc tggccgcatc ttctcaaata tgcttcccag
     4561 cctgcttttc tgtaacgttc accctctacc ttagcatccc ttccctttgc aaatagtcct
     4621 cttccaacaa taataatgtc agatcctgta gagaccacat catccacggt tctatactgt
     4681 tgacccaatg cgtctccctt gtcatctaaa cccacaccgg gtgtcataat caaccaatcg
     4741 taaccttcat ctcttccacc catgtctctt tgagcaataa agccgataac aaaatctttg
     4801 tcgctcttcg caatgtcaac agtaccctta gtatattctc cagtagatag ggagcccttg
     4861 catgacaatt ctgctaacat caaaaggcct ctaggttcct ttgttacttc ttctgccgcc
     4921 tgcttcaaac cgctaacaat acctgggccc accacaccgt gtgcattcgt aatgtctgcc
     4981 cattctgcta ttctgtatac acccgcagag tactgcaatt tgactgtatt accaatgtca
     5041 gcaaattttc tgtcttcgaa gagtaaaaaa ttgtacttgg cggataatgc ctttagcggc
     5101 ttaactgtgc cctccatgga aaaatcagtc aagatatcca catgtgtttt tagtaaacaa
     5161 attttgggac ctaatgcttc aactaactcc agtaattcct tggtggtacg aacatccaat
     5221 gaagcacaca agtttgtttg cttttcgtgc atgatattaa atagcttggc agcaacagga
     5281 ctaggatgag tagcagcacg ttccttatat gtagctttcg acatgattta tcttcgtttc
     5341 ctgcaggttt ttgttctgtg cagttgggtt aagaatactg ggcaatttca tgtttcttca
     5401 acactacata tgcgtatata taccaatcta agtctgtgct ccttccttcg ttcttccttc
     5461 tgttcggaga ttaccgaatc aaaaaaattt caaggaaacc gaaatcaaaa aaaagaataa
     5521 aaaaaaaatg atgaattgaa aagctagctt atcgatgata agctgtcaaa catgagaatt
     5581 aattccacgg actatagact atacctagta tactccgtct actgtacgat acacttccgc
     5641 tcaggtcctt gtcctttaac gaggccttac cactcttttg ttactctatt gatccagctc
     5701 agcaaaggca gtgtgatcta agattctatc ttcgcgatgt agtaaaacta gctagaccga
     5761 gaaagagact agaaatgcaa aaggcacttc tacaatggct gccatcatta ttatccgatg
     5821 tgacgctgca gcttctcaat gatattcgaa tacgctttga ggagatacag cctaatatcc
     5881 gacaaactgt tttacagatt tacgatcgta cttgttaccc atcattgaat tttgaacatc
     5941 cgaacctggg agttttccct gaaacagata gtatatttga acctgtataa taatatatag
     6001 tctagcgctt tacggaagac aatgtatgta tttcggttcc tggagaaact attgcatcta
     6061 ttgcataggt aatcttgcac gtcgcatccc cggttcattt tctgcgtttc catcttgcac
     6121 ttcaatagca tatctttgtt aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca
     6181 acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat
     6241 gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa
     6301 atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca
     6361 gaaatgcaac gcgagagcgc tattttacca acaaagaatc tatacttctt ttttgttcta
     6421 caaaaatgca tcccgagagc gctatttttc taacaaagca tcttagatta ctttttttct
     6481 cctttgtgcg ctctataatg cagtctcttg ataacttttt gcactgtagg tccgttaagg
     6541 ttagaagaag gctactttgg tgtctatttt ctcttccata aaaaaagcct gactccactt
     6601 cccgcgttta ctgattacta gcgaagctgc gggtgcattt tttcaagata aaggcatccc
     6661 cgattatatt ctataccgat gtggattgcg catactttgt gaacagaaag tgatagcgtt
     6721 gatgattctt cattggtcag aaaattatga acggtttctt ctattttgtc tctatatact
     6781 acgtatagga aatgtttaca ttttcgtatt gttttcgatt cactctatga atagttctta
     6841 ctacaatttt tttgtctaaa gagtaatact agagataaac ataaaaaatg tagaggtcga
     6901 gtttagatgc aagttcaagg agcgaaaggt ggatgggtag gttatatagg gatatagcac
     6961 agagatatat agcaaagaga tacttttgag caatgtttgt ggaagcggta ttcgcaatgg
     7021 gaagctccac cccggttgat aatcagaaaa gccccaaaaa caggaagatt gtataagcaa
     7081 atatttaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     7141 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     7201 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     7261 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     7321 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     7381 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     7441 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     7501 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtggggatcg atccactagt
     7561 ccacggatta gaagccgccg agcgggtgac agccctccga aggaagactc tcctccgtgc
     7621 gtcctcgtct tcaccggtcg cgttcctgaa acgcagatgt gcctcgcgcc gcactgctcc
     7681 gaacaataaa gattctacaa tactagcttt ta
//
