LOCUS       sequence-337155-        6249 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 6249)
  AUTHORS   Ochoa-Lizarralde B, Gao YG, Popov AN, Samygina VR, Zhai X, Mishra 
            SK, Boldyrev IA, Molotkovsky JG, Simanshu DK, Patel DJ, Brown RE, 
            Malinina L
  TITLE     Structural analyses of 4-phosphate adaptor protein 2 yield 
            mechanistic insights into sphingolipid recognition by the glycolipid
            transfer protein family.
  JOURNAL   J Biol Chem. 2018 Oct 26;293(43):16709-16723. doi: 
            10.1074/jbc.RA117.000733. Epub 2018 Sep 11.
  PUBMED    30206120
REFERENCE   2  (bases 1 to 6249)
  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: 
            "10.1074/jbc.RA117.000733"; journalName: "J Biol Chem"; date: 
            "2018-10-26- 26"; volume: "293"; issue: "43"; pages: "16709-16723"
COMMENT     Sequence Label: pSUMO FAPP2-GLTPH
FEATURES             Location/Qualifiers
     source          1..6249
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(805..1887)
                     /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     1849..1868
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1888..1965)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2171..2190
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2274..2293
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2274..2292
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2293..2317
                     /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             2332..2354
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2373..2390
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2400..2417
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2430..2723
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             3387..3404
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3457..3475)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3471..3518
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3555..4010
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3642..3661)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3852..3873
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4102..4917)
                     /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(4102..4917)
                     /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     4825..4844
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4909..4928
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5039..5627
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5528..5547
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5781..5798
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5813..5952
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5878..5897)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6038..6060
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6054..6245)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg
       61 atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt
      121 aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag
      181 cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag
      241 atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg
      301 aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt
      361 cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct
      421 agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc catgccggcg
      481 ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg
      541 agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga
      601 aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg
      661 ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag
      721 ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc
      781 taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
      841 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag
      901 ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc
      961 ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt
     1021 gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac
     1081 cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc
     1141 catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat
     1201 ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat
     1261 ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact
     1321 taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc
     1381 cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac
     1441 atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc
     1501 atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc
     1561 cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag
     1621 ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact
     1681 ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt
     1741 gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac
     1801 gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc
     1861 gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg
     1921 ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct
     1981 ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca
     2041 ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg
     2101 ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat
     2161 cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga
     2221 tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga aattaatacg
     2281 actcactata ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa
     2341 ctttaagaag gagatatacc atgggcagca gccatcatca tcatcatcac agcagcggcc
     2401 tggtgccgcg cggcagccat atggctagca tgtcggactc agaagtcaat caagaagcta
     2461 agccagaggt caagccagaa gtcaagcctg agactcacat caatttaaag gtgtccgatg
     2521 gatcttcaga gatcttcttc aagatcaaaa agaccactcc tttaagaagg ctgatggaag
     2581 cgttcgctaa aagacagggt aaggaaatgg actccttaag attcttgtac gacggtatta
     2641 gaatccaagc tgatcagacc cctgaagatt tggacatgga ggataacgat attattgagg
     2701 ctcacagaga acagattggt ggatccactt tctttagtac catgaacaca agctttagtg
     2761 acattgaact tctggaagac agtggcattc ccacagaagc attcttggca tcatgttatg
     2821 ctgtggttcc agtattagac aaacttggcc ctacagtgtt tgctcctgtt aagatggatc
     2881 ttgttggaaa tattaagaaa gtaaatcaga agtatataac caacaaagaa gagtttacca
     2941 ctctccagaa gatagtgctg cacgaagtgg aggcggatgt agcccaggtt aggaactcag
     3001 cgactgaagc cctcttgtgg ctgaagagag gtctcaaatt tttgaaggga tttttgacag
     3061 aagtgaaaaa tggggagaag gatatccaga cagccctaaa taatgcatat ggtaaaacat
     3121 tgcggcaaca ccatggctgg gtagttcgag gggtttttgc gttagcttta agggcagctc
     3181 catcctatga agattttgtg gccgcgttaa ccgtaaagga aggtgaccac cagaaagaag
     3241 ctttcagtat tgggatgcag agggacctca gcctttacct ccctgccatg gagaagcagc
     3301 tggccatact ggacacttta tatgaggtcc acgggctgga atctgatgag gtggtatgag
     3361 tcgacaagct tgcggccgca ctcgagcacc accaccacca ccactgagat ccggctgcta
     3421 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
     3481 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg
     3541 gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3601 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3661 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3721 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3781 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3841 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     3901 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     3961 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
     4021 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4081 tatccgctca tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt
     4141 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
     4201 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
     4261 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
     4321 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt
     4381 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
     4441 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg
     4501 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
     4561 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc
     4621 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
     4681 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
     4741 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat
     4801 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     4861 catgttggaa tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcataac
     4921 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgacc aaaatccctt
     4981 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5041 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5101 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5161 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     5221 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5281 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5341 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5401 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5461 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5521 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5581 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5641 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5701 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     5761 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     5821 ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac
     5881 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
     5941 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     6001 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     6061 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
     6121 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
     6181 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
     6241 gtcactgat
//
