LOCUS       sequence-337152-        4533 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 4533)
  AUTHORS   Hirano Y, Gao YG, Stephenson DJ, Vu NT, Malinina L, Simanshu DK, 
            Chalfant CE, Patel DJ, Brown RE
  TITLE     Structural basis of phosphatidylcholine recognition by the C2-domain
            of cytosolic phospholipase A2alpha.
  JOURNAL   Elife. 2019 May 3;8. pii: 44760. doi: 10.7554/eLife.44760.
  PUBMED    31050338
REFERENCE   2  (bases 1 to 4533)
  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: "Elife."; 
            date: "2019-05-3"; volume: "8. pii: 44760. doi"; pages: " 
            10.7554/eLife.44760"
COMMENT     Sequence Label: pSUMO chicken-cPLA2-C2-domain
FEATURES             Location/Qualifiers
     source          1..4533
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(71..1153)
                     /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     1115..1134
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1154..1231)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        1277..1295
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1296..1320
                     /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             1376..1393
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1403..1420
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1433..1726
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     promoter        2211..2229
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2230..2254
                     /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             2363..2407
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2445..2463)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2459..2506
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2736..3551)
                     /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(2736..3551)
                     /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     3459..3478
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3552..3643)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3659..4408)
                     /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(3669..3686)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3909..3928)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat
       61 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
      121 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
      181 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
      241 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
      301 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac
      361 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
      421 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
      481 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
      541 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
      601 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
      661 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
      721 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
      781 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
      841 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
      901 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
      961 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
     1021 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
     1081 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
     1141 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     1201 cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac
     1261 tcctgcatta ggaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc
     1321 cctgtagaaa taattttgtt taactttaat aaggagatat accatgggca gcagccatca
     1381 tcatcatcat cacagcagcg gcctggtgcc gcgcggcagc cacatggcaa gcatgtcgga
     1441 ctcagaagtc aatcaagaag ctaagccaga ggtcaagcca gaagtcaagc ctgagactca
     1501 catcaattta aaggtgtccg atggatcttc agagatcttc ttcaagatca aaaagaccac
     1561 tcctttaaga aggctgatgg aagcgttcgc taaaagacag ggtaaggaaa tggactcctt
     1621 aagattcttg tacgacggta ttagaatcca agctgatcag acccctgaag atttggacat
     1681 ggaggataac gatattattg aggctcacag agaacagatt ggtggatcct atagccatgt
     1741 gtttaccgtg accgtgcgca aagccaccaa tgtgaccaaa ggcgccattg gcgatatgct
     1801 ggatacccct gatccttatg tggagctgtt tattccgagc gcaccggatt gccgcaaacg
     1861 caccaagcac ttcaacaacg acgtgaaccc ggtgtggaac gagaccttcg agtttatcct
     1921 ggacccgaac caggataacg tgctggaagt gaccctgatg gacgccaact atgtgatgga
     1981 tgagaccctg ggcatggcca cctttccgat tagcagcctg aaactgggcg agaagaaaga
     2041 agttcagctg accttcaaca acgtgaccga gatgaccctg gaactgagcc tggaagtttg
     2101 cagctgatga attcgagctc ggcgcgcctg caggtcgaca agcttgcggc cgcataatgc
     2161 ttaagtcgaa cagaaagtaa tcgtattgta cacggccgca taatcgaaat taatacgact
     2221 cactataggg gaattgtgag cggataacaa ttccccatct tagtatatta gttaagtata
     2281 agaaggagat atacatatgg cagatctcaa ttggatatcg gccggccacg cgatcgctga
     2341 cgtcggtacc ctcgagtctg gtaaagaaac cgctgctgcg aaatttgaac gccagcacat
     2401 ggactcgtct actagcgcag cttaattaac ctaggctgct gccaccgctg agcaataact
     2461 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aacctcaggc
     2521 atttgagaag cacacggtca cactgcttcc ggtagtcaat aaaccggtaa accagcaata
     2581 gacataagcg gctatttaac gaccctgccc tgaaccgacg acaagctgac gaccgggtct
     2641 ccgcaagtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2701 tacattcaaa tatgtatccg ctcatgaatt aattcttaga aaaactcatc gagcatcaaa
     2761 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     2821 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     2881 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     2941 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt
     3001 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     3061 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcgg
     3121 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     3181 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     3241 ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     3301 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     3361 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     3421 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     3481 tataaatcag catccatgtt ggaatttaat cgcggcctag agcaagacgt ttcccgttga
     3541 atatggctca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     3601 atgagcggat acatatttga atgtatttag aaaaataaac aaataggcat gcagcgctct
     3661 tccgcttcct cgctcactga ctcgctacgc tcggtcgttc gactgcggcg agcggtgtca
     3721 gctcactcaa aagcggtaat acggttatcc acagaatcag gggataaagc cggaaagaac
     3781 atgtgagcaa aaagcaaagc accggaagaa gccaacgccg caggcgtttt tccataggct
     3841 ccgcccccct gacgagcatc acaaaaatcg acgctcaagc cagaggtggc gaaacccgac
     3901 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3961 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4021 tcatagctca cgctgttggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4081 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4141 gtccaacccg gtaagacacg acttatcgcc actggcagca gccattggta actgatttag
     4201 aggactttgt cttgaagtta tgcacctgtt aaggctaaac tgaaagaaca gattttggtg
     4261 agtgcggtcc tccaacccac ttaccttggt tcaaagagtt ggtagctcag cgaaccttga
     4321 gaaaaccacc gttggtagcg gtggtttttc tttatttatg agatgatgaa tcaatcggtc
     4381 tatcaagtca acgaacagct attccgttac tctagatttc agtgcaattt atctcttcaa
     4441 atgtagcacc tgaagtcagc cccatacgat ataagttgta attctcatgt tagtcatgcc
     4501 ccgcgcccac cggaaggagc tgactgggtt gaa
//
