LOCUS       sequence-337024-        4557 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The plasmids contain inserts (ORFs) for expressing various members 
            of the GlycoLipid Transfer Protein (GLTP) superfamily..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4557)
  AUTHORS   Simanshu DK, Kamlekar RK, Wijesinghe DS, Zou X, Zhai X, Mishra SK, 
            Molotkovsky JG, Malinina L, Hinchcliffe EH, Chalfant CE, Brown RE, 
            Patel DJ
  TITLE     Non-vesicular trafficking by a ceramide-1-phosphate transfer protein
            regulates eicosanoids.
  JOURNAL   Nature. 2013 Aug 22;500(7463):463-7. doi: 10.1038/nature12332. Epub 
            2013 Jul 17.
  PUBMED    23863933
REFERENCE   2  (bases 1 to 4557)
  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"; doi: 
            "10.1038/nature12332"; journalName: "Nature"; date: "2013-08-22- 
            22"; volume: "500"; issue: "7463"; pages: "463-7"
COMMENT     Sequence Label: pSUMO K60A-CPTP
FEATURES             Location/Qualifiers
     source          1..4557
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(75..1157)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1119..1138
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1158..1235)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1281..1300
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1281..1299
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1300..1324
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             1380..1397
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1407..1424
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1437..1730
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     regulatory      1975..1984
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2391..2435
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2473..2491)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2487..2534
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2764..3579)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2764..3579)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3487..3506
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3580..3671)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3687..4436)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(3697..3714)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3937..3956)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat
       61 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
      121 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt
      181 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt
      241 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt
      301 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatgtcc
      361 gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg
      421 ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga
      481 aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga
      541 gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc
      601 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta
      661 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat
      721 aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga
      781 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag
      841 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg
      901 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca
      961 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa
     1021 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc
     1081 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat
     1141 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc
     1201 ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg
     1261 cgactcctgc attaggaaat taatacgact cactataggg gaattgtgag cggataacaa
     1321 ttcccctgta gaaataattt tgtttaactt taataaggag atataccatg ggcagcagcc
     1381 atcatcatca tcatcacagc agcggcctgg tgccgcgcgg cagccacatg gcaagcatgt
     1441 cggactcaga agtcaatcaa gaagctaagc cagaggtcaa gccagaagtc aagcctgaga
     1501 ctcacatcaa tttaaaggtg tccgatggat cttcagagat cttcttcaag atcaaaaaga
     1561 ccactccttt aagaaggctg atggaagcgt tcgctaaaag acagggtaag gaaatggact
     1621 ccttaagatt cttgtacgac ggtattagaa tccaagctga tcagacccct gaagatttgg
     1681 acatggagga taacgatatt attgaggctc acagagaaca gattggtgga tccatggatg
     1741 actcggagac aggtttcaat ctgaaagtcg tcctggtcag tttcaagcag tgtctcgatg
     1801 agaaggaaga ggtcttgctg gacccctaca ttgccagctg gaagggcctg gtcaggtttc
     1861 tgaacagcct gggcaccatc ttctcattca tctccaagga cgtggtctcc gcgctgcgga
     1921 tcatggagcg cctcaggggc ggcccgcaga gcgagcacta ccgcagcctg caggccatgg
     1981 tggcccatga gctgagcaac cggctggtgg acctggagcg ccgctcccac cacccggagt
     2041 ctggctgccg gacggtgctg cgcctgcacc gcgccctgca ctggctgcag ctgttcctgg
     2101 agggcctgcg taccagcccc gaggacgcac gcacctccgc gctctgcgcc gactcctaca
     2161 acgcctcgct ggccgcctac cacccctggg ttgtgcgccg cgccgtcacc gtggccttct
     2221 gcacgctgcc cacacgcgag gtcttcctgg aggccatgaa cgtggggccc ccggagcagg
     2281 ccgtgcagat gctaggcgag gccctcccct tcatccagcg tgtctacaac gtctcccaga
     2341 agctctacgc cgagcactcc ctgctggacc tgccctaggt cgagtctggt aaagaaaccg
     2401 ctgctgcgaa atttgaacgc cagcacatgg actcgtctac tagcgcagct taattaacct
     2461 aggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
     2521 tgaggggttt tttgctgaaa cctcaggcat ttgagaagca cacggtcaca ctgcttccgg
     2581 tagtcaataa accggtaaac cagcaataga cataagcggc tatttaacga ccctgccctg
     2641 aaccgacgac aagctgacga ccgggtctcc gcaagtggca cttttcgggg aaatgtgcgc
     2701 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgaattaa
     2761 ttcttagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
     2821 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
     2881 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
     2941 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
     3001 gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg
     3061 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
     3121 ttgcgcctga gcgagacgaa atacgcggtc gctgttaaaa ggacaattac aaacaggaat
     3181 cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg
     3241 atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc
     3301 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
     3361 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
     3421 aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc
     3481 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
     3541 cggcctagag caagacgttt cccgttgaat atggctcata ctcttccttt ttcaatatta
     3601 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     3661 aaataaacaa ataggcatgc agcgctcttc cgcttcctcg ctcactgact cgctacgctc
     3721 ggtcgttcga ctgcggcgag cggtgtcagc tcactcaaaa gcggtaatac ggttatccac
     3781 agaatcaggg gataaagccg gaaagaacat gtgagcaaaa agcaaagcac cggaagaagc
     3841 caacgccgca ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3901 gctcaagcca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3961 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4021 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgttggtat ctcagttcgg
     4081 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4141 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4201 tggcagcagc cattggtaac tgatttagag gactttgtct tgaagttatg cacctgttaa
     4261 ggctaaactg aaagaacaga ttttggtgag tgcggtcctc caacccactt accttggttc
     4321 aaagagttgg tagctcagcg aaccttgaga aaaccaccgt tggtagcggt ggtttttctt
     4381 tatttatgag atgatgaatc aatcggtcta tcaagtcaac gaacagctat tccgttactc
     4441 tagatttcag tgcaatttat ctcttcaaat gtagcacctg aagtcagccc catacgatat
     4501 aagttgtaat tctcatgtta gtcatgcccc gcgcccaccg gaaggagctg actgggt
//
