LOCUS       sequence-337151-        5994 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 5994)
  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 5994)
  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 human-cPLA2-C2-domain
FEATURES             Location/Qualifiers
     source          1..5994
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(37..1119)
                     /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     1081..1100
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1120..1197)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1403..1422
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1506..1525
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1506..1524
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1525..1549
                     /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)."
     RBS             1564..1586
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1605..1622
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1632..1649
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1662..1955
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEVKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             2364..2381
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2434..2452)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2448..2495
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2532..2987
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2619..2638)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2829..2850
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3079..3894)
                     /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(3079..3894)
                     /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     3802..3821
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3886..3905
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4016..4604
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4505..4524
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4758..4775
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4790..4929
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4855..4874)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5015..5037
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5031..5222)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
       61 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
      121 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
      181 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
      241 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
      301 gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
      361 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
      421 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
      481 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
      541 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
      601 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
      661 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
      721 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
      781 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
      841 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
      901 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
      961 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
     1021 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
     1081 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
     1141 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
     1201 gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga
     1261 ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc
     1321 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
     1381 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
     1441 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc
     1501 gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc ctctagaaat
     1561 aattttgttt aactttaaga aggagatata ccatgggcag cagccatcat catcatcatc
     1621 acagcagcgg cctggtgccg cgcggcagcc atatggctag catgtcggac tcagaagtca
     1681 atcaagaagt taagccagag gtcaagccag aagtcaagcc tgagactcac atcaatttaa
     1741 aggtgtccga tggatcttca gagatcttct tcaagatcaa aaagaccact cctttaagaa
     1801 ggctgatgga agcgttcgct aaaagacagg gtaaggaaat ggactcctta agattcttgt
     1861 acgacggtat tagaatccaa gctgatcaga cccctgaaga tttggacatg gaggataacg
     1921 atattattga ggctcacaga gaacagattg gtggatccgg atcccacaag tttacggtag
     1981 tggtgttacg tgccaccaaa gtgacaaagg gggcctttgg tgacatgctt gatactccag
     2041 atccctatgt ggaacttttt atctctacaa cccctgacag caggaagaga acaagacatt
     2101 tcaataatga cataaaccct gtgtggaatg agacctttga atttattttg gatcctaatc
     2161 aggaaaatgt tttggagatt acgttaatgg atgccaatta tgtcatggat gaaactctag
     2221 ggacagcaac atttactgta tcttctatga aggtgggaga aaagaaagaa gttcctttta
     2281 ttttcaacca agtcactgaa atggttctag aaatgtctct tgaagtttgc tcataggtcg
     2341 acaagcttgc ggccgcactc gagcaccacc accaccacca ctgagatccg gctgctaaca
     2401 aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc
     2461 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat
     2521 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2581 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     2641 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     2701 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     2761 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     2821 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     2881 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     2941 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     3001 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3061 ccgctcatga attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
     3121 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     3181 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     3241 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     3301 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
     3361 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     3421 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
     3481 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     3541 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
     3601 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     3661 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     3721 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
     3781 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     3841 gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcataacacc
     3901 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgaccaaa atcccttaac
     3961 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4021 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4081 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4141 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     4201 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4261 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4321 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4381 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     4441 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4501 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4561 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     4621 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     4681 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     4741 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     4801 attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat
     4861 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
     4921 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     4981 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     5041 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
     5101 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
     5161 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
     5221 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
     5281 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
     5341 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
     5401 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
     5461 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
     5521 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
     5581 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
     5641 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ggccgccatg
     5701 ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct
     5761 tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc
     5821 cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt
     5881 tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg
     5941 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcc
//
