LOCUS       sequence-338659-        6077 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 6077)
  AUTHORS   Malinina L, Malakhova ML, Teplov A, Brown RE, Patel DJ
  TITLE     Structural basis for glycosphingolipid transfer specificity.
  JOURNAL   Nature. 2004 Aug 26;430(7003):1048-53. doi: 10.1038/nature02856.
  PUBMED    15329726
REFERENCE   2  (bases 1 to 6077)
  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"; journalName: 
            "Nature"; date: "2004-08-26- 26"; volume: "430"; issue: "7003"; 
            pages: "1048-53"
COMMENT     Sequence Label: pET30 GLTP
FEATURES             Location/Qualifiers
     source          1..6077
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     563..582
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(602..679)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     885..904
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     992..1011
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        992..1010
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1011..1035
                     /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             1050..1072
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1083..1100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1110..1127
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1134..1178
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             1209..1220
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1929..1946
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1999..2017)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2013..2060
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2097..2552
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2184..2203)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2394..2415
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2644..3459)
                     /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(2644..3459)
                     /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     3367..3386
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3451..3470
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3581..4169
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4070..4089
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4323..4340
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4355..4494
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4420..4439)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4580..4602
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4596..4787)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5596..6077,1..601))
                     /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"
ORIGIN
        1 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
       61 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
      121 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
      181 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
      241 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
      301 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
      361 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
      421 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
      481 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
      541 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
      601 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
      661 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
      721 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
      781 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
      841 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
      901 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
      961 aggatcgaga tcgatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga
     1021 gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga gatatacata
     1081 tgcaccatca tcatcatcat tcttctggtc tggtgccacg cggttctggt atgaaagaaa
     1141 ccgctgctgc taaattcgaa cgccagcaca tggacagccc agatctgggt accggtggtg
     1201 gctccggtat tgagggtcgc atggcgctgc tggcggagca cttgctgaag ccgctgcccg
     1261 cggacaagca gatcgagacc gggcccttcc tcgaggcggt gtcccacctg ccgcccttct
     1321 tcgattgcct tgggtcccca gtgtttactc ccatcaaggc agacataagc ggcaacatca
     1381 cgaaaatcaa agctgtgtac gacaccaacc cagccaagtt ccggaccctg cagaacatcc
     1441 tggaggtgga gaaagaaatg tatggagcag agtggcccaa agtaggggcc acactggcgc
     1501 tgatgtggct gaaaagaggc ctccgcttca tccaggtctt cctccagagc atctgcgacg
     1561 gggagcggga cgagaaccac cccaacctca tccgtgtcaa cgccaccaag gcctacgaga
     1621 tggccctcaa gaagtaccat ggctggatcg tgcagaagat cttccaggca gcactgtacg
     1681 cagcacccta taagtctgac ttcctgaaag cgctctccaa ggggcagaat gttacggagg
     1741 aggagtgcct ggagaagatc cgcctcttcc tagtcaacta cacggcgacc atcgatgtca
     1801 tctacgagat gtacacccag atgaacgccg agcttaacta caaggtgtaa ggctctaact
     1861 ctcctctggc catggcgata tcggatccga attcgagctc cgtcgacaag cttgcggccg
     1921 cactcgagca ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag
     1981 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
     2041 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc
     2101 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2161 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2221 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     2281 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     2341 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2401 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     2461 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     2521 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
     2581 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgaattaa
     2641 ttcttagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
     2701 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
     2761 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
     2821 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
     2881 gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg
     2941 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
     3001 ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
     3061 cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg
     3121 atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc
     3181 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
     3241 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
     3301 aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc
     3361 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
     3421 cggcctagag caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt
     3481 tatgtaagca gacagtttta ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc
     3541 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3601 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3661 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3721 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3781 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3841 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3901 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3961 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4021 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     4081 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     4141 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     4201 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     4261 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     4321 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     4381 atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc
     4441 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     4501 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     4561 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     4621 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
     4681 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     4741 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     4801 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
     4861 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
     4921 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     4981 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     5041 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     5101 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
     5161 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
     5221 ctcaacgaca ggagcacgat catgcgcacc cgtggggccg ccatgccggc gataatggcc
     5281 tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc
     5341 aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc
     5401 tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag
     5461 acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg
     5521 ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca
     5581 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5641 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt
     5701 tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag
     5761 ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt
     5821 taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatgtc
     5881 cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc
     5941 gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg
     6001 aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg
     6061 agtgagatat ttatgcc
//
