LOCUS       sequence-337027-        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 R106L-CPTP
FEATURES             Location/Qualifiers
     source          1..4557
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(55..1137)
                     /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     1099..1118
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1138..1215)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1261..1280
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1261..1279
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1280..1304
                     /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             1360..1377
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1387..1404
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1417..1710
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     regulatory      1955..1964
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2371..2415
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2453..2471)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2467..2514
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2744..3559)
                     /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(2744..3559)
                     /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     3467..3486
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3560..3651)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3667..4416)
                     /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(3677..3694)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3917..3936)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
       61 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
      121 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
      181 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
      241 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
      301 gctgtcttcg gtatcgtcgt atcccactac cgagatgtcc gcaccaacgc gcagcccgga
      361 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
      421 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
      481 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
      541 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
      601 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
      661 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
      721 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
      781 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
      841 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
      901 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
      961 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     1021 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     1081 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     1141 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg
     1201 ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaaat
     1261 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
     1321 tgtttaactt taataaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
     1381 agcggcctgg tgccgcgcgg cagccacatg gcaagcatgt cggactcaga agtcaatcaa
     1441 gaagctaagc cagaggtcaa gccagaagtc aagcctgaga ctcacatcaa tttaaaggtg
     1501 tccgatggat cttcagagat cttcttcaag atcaaaaaga ccactccttt aagaaggctg
     1561 atggaagcgt tcgctaaaag acagggtaag gaaatggact ccttaagatt cttgtacgac
     1621 ggtattagaa tccaagctga tcagacccct gaagatttgg acatggagga taacgatatt
     1681 attgaggctc acagagaaca gattggtgga tccatggatg actcggagac aggtttcaat
     1741 ctgaaagtcg tcctggtcag tttcaagcag tgtctcgatg agaaggaaga ggtcttgctg
     1801 gacccctaca ttgccagctg gaagggcctg gtcaggtttc tgaacagcct gggcaccatc
     1861 ttctcattca tctccaagga cgtggtctcc aagctgcgga tcatggagcg cctcaggggc
     1921 ggcccgcaga gcgagcacta ccgcagcctg caggccatgg tggcccatga gctgagcaac
     1981 cggctggtgg acctggagcg ccgctcccac cacccggagt ctggctgcct gacggtgctg
     2041 cgcctgcacc gcgccctgca ctggctgcag ctgttcctgg agggcctgcg taccagcccc
     2101 gaggacgcac gcacctccgc gctctgcgcc gactcctaca acgcctcgct ggccgcctac
     2161 cacccctggg ttgtgcgccg cgccgtcacc gtggccttct gcacgctgcc cacacgcgag
     2221 gtcttcctgg aggccatgaa cgtggggccc ccggagcagg ccgtgcagat gctaggcgag
     2281 gccctcccct tcatccagcg tgtctacaac gtctcccaga agctctacgc cgagcactcc
     2341 ctgctggacc tgccctaggt cgagtctggt aaagaaaccg ctgctgcgaa atttgaacgc
     2401 cagcacatgg actcgtctac tagcgcagct taattaacct aggctgctgc caccgctgag
     2461 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
     2521 cctcaggcat ttgagaagca cacggtcaca ctgcttccgg tagtcaataa accggtaaac
     2581 cagcaataga cataagcggc tatttaacga ccctgccctg aaccgacgac aagctgacga
     2641 ccgggtctcc gcaagtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2701 tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga
     2761 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     2821 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     2881 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     2941 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     3001 gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     3061 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
     3121 atacgcggtc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     3181 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     3241 atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     3301 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
     3361 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     3421 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     3481 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt
     3541 cccgttgaat atggctcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     3601 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggcatgc
     3661 agcgctcttc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag
     3721 cggtgtcagc tcactcaaaa gcggtaatac ggttatccac agaatcaggg gataaagccg
     3781 gaaagaacat gtgagcaaaa agcaaagcac cggaagaagc caacgccgca ggcgtttttc
     3841 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagcca gaggtggcga
     3901 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3961 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4021 gcgctttctc atagctcacg ctgttggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4081 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4141 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cattggtaac
     4201 tgatttagag gactttgtct tgaagttatg cacctgttaa ggctaaactg aaagaacaga
     4261 ttttggtgag tgcggtcctc caacccactt accttggttc aaagagttgg tagctcagcg
     4321 aaccttgaga aaaccaccgt tggtagcggt ggtttttctt tatttatgag atgatgaatc
     4381 aatcggtcta tcaagtcaac gaacagctat tccgttactc tagatttcag tgcaatttat
     4441 ctcttcaaat gtagcacctg aagtcagccc catacgatat aagttgtaat tctcatgtta
     4501 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatc
//
