LOCUS       sequence-337075-        6880 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 6880)
  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 6880)
  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: pFLAG K60A-CPTP
FEATURES             Location/Qualifiers
     source          1..6880
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             945..968
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      1228..1237
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1639..2261
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1838..1857)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2285..2305)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2303..2600
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2654..3448
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2708..2727)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3318..3337
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4100..4234
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4137..4156)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4191..4210
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4239..4258)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4240..4258)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4272..4288)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4272..4288)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4285..4307)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4296..4312
                     /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)."
     promoter        complement(join(4320..4326,4327..4344,4345..4350))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4365..4386
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4503..4520)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4674..5262)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4754..4773)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5433..6224,6225..6293))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6056..6075
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6294..6398)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6425..6853
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6512..6531)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6722..6743
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaattcag atcgatctac catggactac aaagacgatg
      961 acgacaagct tgcggccgcg aattcaatgg atgactcgga gacaggtttc aatctgaaag
     1021 tcgtcctggt cagtttcaag cagtgtctcg atgagaagga agaggtcttg ctggacccct
     1081 acattgccag ctggaagggc ctggtcaggt ttctgaacag cctgggcacc atcttctcat
     1141 tcatctccaa ggacgtggtc tccgcgctgc ggatcatgga gcgcctcagg ggcggcccgc
     1201 agagcgagca ctaccgcagc ctgcaggcca tggtggccca tgagctgagc aaccggctgg
     1261 tggacctgga gcgccgctcc caccacccgg agtctggctg ccggacggtg ctgcgcctgc
     1321 accgcgccct gcactggctg cagctgttcc tggagggcct gcgtaccagc cccgaggacg
     1381 cacgcacctc cgcgctctgc gccgactcct acaacgcctc gctggccgcc taccacccct
     1441 gggttgtgcg ccgcgccgtc accgtggcct tctgcacgct gcccacacgc gaggtcttcc
     1501 tggaggccat gaacgtgggg cccccggagc aggccgtgca gatgctaggc gaggccctcc
     1561 ccttcatcca gcgtgtctac aacgtctccc agaagctcta cgccgagcac tccctgctgg
     1621 acctgcccta gggatcccgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc
     1681 ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt
     1741 ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag
     1801 gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt
     1861 gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg
     1921 cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg
     1981 tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct
     2041 caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct
     2101 cccttccctg tccttctgat tttaaaataa ctataccagc aggaggacgt ccagacacag
     2161 cataggctac ctggccatgc ccaaccggtg ggacatttga gttgcttgct tggcactgtc
     2221 ctctcatgcg ttgggtccac tcagtagatg cctgttgaat tgggtacgcg gccagcttgg
     2281 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     2341 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2401 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2461 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2521 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     2581 tagtgaggag gcttttttgg aggaattgat cagcttggga tctgatcaag agacaggatg
     2641 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     2701 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     2761 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     2821 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     2881 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     2941 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     3001 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     3061 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     3121 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     3181 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     3241 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     3301 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     3361 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     3421 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     3481 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     3541 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     3601 ctggagttct tcgcccaccc cgggctcgat cccctcgcga gttggttcag ctgctgcctg
     3661 aggctggacg acctcgcgga gttctaccgg cagtgcaaat ccgtcggcat ccaggaaacc
     3721 agcagcggct atccgcgcat ccatgccccc gaactgcagg agtggggagg cacgatggcc
     3781 gctttggtcg acccggacgg gacgctcctg cgcctgatac agaacgaatt gcttgcaggc
     3841 atctcatgag tgtgtcttcc cgttttccgc ctgaggtcac tgcgtggatg gagcgctggc
     3901 gcctgctgcg cgacggcgag ctgctcacca cccactcgcc aagctggaac cgtaaaaagg
     3961 ccgcgttgct ggcgtttttc cataggctcc gccgatcata atcagccata ccacatttgt
     4021 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     4081 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     4141 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4201 caaactcatc aatgtatctt atcatgtctg gatcaattcc ctatagtgag tcgtattaaa
     4261 ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4321 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4381 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4441 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4501 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4561 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4621 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4681 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4741 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4801 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4861 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4921 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4981 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5041 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5101 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5161 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5221 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5281 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5341 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5401 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5461 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5521 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5581 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5641 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5701 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5761 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5821 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     5881 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5941 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6001 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6061 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6121 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6181 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6241 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6301 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6361 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6421 cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     6481 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     6541 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     6601 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     6661 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     6721 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     6781 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     6841 tttaacgcga attttaacaa aatattaacg cttacaattt
//
