LOCUS       sequence-446871-        5412 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human MOSPD3 fused to EGFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5412)
  AUTHORS   Eichler J, Huver S, Knorr CJ, Wendling C, Kobayashi T, Tomasetto C, 
            Alpy F
  TITLE     Methods for Visualizing and Quantifying Cholesterol Distribution in 
            Mammalian Cells Using Filipin and D4 Probes.
  JOURNAL   Methods Mol Biol. 2025;2888:101-118. doi: 
            10.1007/978-1-0716-4318-1_8.
  PUBMED    39699727
REFERENCE   2  (bases 1 to 5412)
  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: "Methods Mol
            Biol."; date: "2025"; volume: "2888:101-118. doi"; pages: " 
            10.1007/978-1-0716-4318-1_8"
COMMENT     Sequence Label: pEGFPC1 MOSPD3
FEATURES             Location/Qualifiers
     source          1..5412
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        57..360
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        361..564
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     515..535
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      603..612
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(609..611,612..614,615..1325)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(609..611,612..614,615..1325)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(654..675)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(915..934)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1262..1283
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2196..2317
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2233..2252)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2287..2306
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2324..2779)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2461..2482)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2673..2692
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2806..2910
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2912..3269
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2912..3103
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2935..2955)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3120..3255
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3304..4098
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3304..4098
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3358..3377)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3968..3987
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4286..4305)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4330..4377
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4706..5294
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5195..5214
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
       61 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      121 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      181 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      241 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      301 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      361 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      421 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      481 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      541 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      601 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      661 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      721 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
      781 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
      841 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
      901 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      961 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     1021 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     1081 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     1141 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     1201 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     1261 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     1321 acaagtccgg actcagatct cgagctcaag cttcgaattg tcgccgtggg gcgccccagg
     1381 accaggagct ggtgggtccg gggccccctg ggcgggggtc ccggggcgcc cctcctccct
     1441 tgggacccgt tgtcccggtc ctggtctttc ccccggatct agtattcagg gcggaccagc
     1501 ggagcggacc ccgacagctg ctgaccctct ataaccccac aggaactgcg cttcgcttcc
     1561 gagtcctgtg cacagcacct gccaaataca cggtgtttga cgcagaggga tatgtgaagc
     1621 cccagtcttg cattgacatt gtgattcgcc atgtggcacc cattcccagc cactatgatg
     1681 tccaggaccg cttccgcatt gagctgtctg aggaaggagc tgagggccga gtggtgggac
     1741 gcaaggacat tacctccatt ctgagagccc cagcgtaccc ccttgagctt cagggacagc
     1801 cagatccagc gcctcgccca gggcctcctg ctgggacacc accacccacg gccagacact
     1861 tccaggagca cccccgccag caactggcca ccagctcctt cctcctcttc ttgctgacgg
     1921 ggattgtgtc tgtggccttc ctgctgctcc cactcccgga cgaactcggc agccagctgc
     1981 ctcaagtcct gcacgtctcc ctgggacaaa agttggtggc cgcctacgtc ttgggcctcc
     2041 tcaccatggt gttcctccgg acctgaggat ccaccggatc tagataactg atcataatca
     2101 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     2161 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     2221 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     2281 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat
     2341 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     2401 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     2461 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     2521 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     2581 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     2641 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     2701 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     2761 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     2821 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2881 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     2941 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3001 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3061 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3121 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3181 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3241 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     3301 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3361 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3421 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3481 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3541 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3601 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3661 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3721 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3781 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     3841 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     3901 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     3961 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4021 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4081 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4141 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4201 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4261 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     4321 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     4381 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     4441 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     4501 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     4561 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4621 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4681 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4741 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4801 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4861 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4921 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4981 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5041 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5101 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5161 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5221 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5281 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5341 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     5401 ccatgcatta gt
//
