LOCUS       sequence-433907-        8399 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Golgi-targeted LOVPLD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8399)
  AUTHORS   Li XL, Tei R, Uematsu M, Baskin JM
  TITLE     Ultralow Background Membrane Editors for Spatiotemporal Control of 
            Phosphatidic Acid Metabolism and Signaling.
  JOURNAL   ACS Cent Sci. 2024 Jan 30;10(3):543-554. doi: 
            10.1021/acscentsci.3c01105. eCollection 2024 Mar 27.
  PUBMED    38559292
REFERENCE   2  (bases 1 to 8399)
  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: 
            "10.1021/acscentsci.3c01105"; journalName: "ACS Cent Sci"; date: 
            "2024-01-30- 30"; volume: "10"; issue: "3"; pages: "543-554"
COMMENT     Sequence Label: pCDNA3-LOVPLD-mCherry-OSBP(PH)
FEATURES             Location/Qualifiers
     source          1..8399
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      895..904
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2896..3600
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(3038..3056)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3306..3325
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        complement(3952..3970)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3953..3970)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3990..4007)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3996..4220
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4266..4694
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4353..4372)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4563..4584
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4703..4723)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4708..5037
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4888..5023
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4950..4969
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5104..5898
                     /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(5158..5177)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5768..5787
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6072..6193
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6109..6128)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6163..6182
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6242..6258)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6242..6258)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6255..6277)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6266..6282
                     /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(6290..6296,6297..6314,6315..6320))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6335..6356
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6473..6490)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6644..7232)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6724..6743)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7403..8194,8195..8263))
                     /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     8026..8045
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8264..8368)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgccacc
      901 atgggatccg cagattcagc tacaccacat ttggacgccg ttgagcaaac attgaggcaa
      961 gtgtccccag gcctggaggg cgatgtatgg gaaagaacta gtggaaataa attggacgga
     1021 agtgccgctg atcccagtga ttggttgttg caaaccccag gatgttgggg cgatgataga
     1081 tgcgttgaca gagttgggac aaaaaggttg ctggctaaga tgactgaaaa tatagggaat
     1141 gccaccagaa ccgtagatat tagtacactt gcacctttcc caaacggcgc attccaagac
     1201 gctattgtcg ctggtttgaa ggagtccgca gctagaggca ataagttgaa agttaggata
     1261 ctggttgggg ctgcacccgt ctaccacatg aatgtaatac cttccaagta tagggacgaa
     1321 ttgactgcaa aacttggcaa agccgcagaa aacataactt tgaacgtggc tagtatgact
     1381 acatcaaaaa cagctttctc ctggaaccat agcaaaatct tggttgtaga cgggcaaagc
     1441 gcccttactg gtggaataaa tagctggaag gatgattact tggatactac tcatccagtg
     1501 tctgatgttg atcttgcact gaccggacca gctgcaggct ctgccggtag ataccttgac
     1561 actttgtgga catggacctg ccaaaacaag agtaacatcg cttccgtctg gtttgcagct
     1621 tccggcaacg cagggtgcat ggccactatg cataaagata caaaccccaa ggcatcacct
     1681 gccaccggga atgtcccaat aatagccgtt ggagggttgg gcgttggcat caaagatgtg
     1741 gaccccaagt ccactttcag gcccgatctg ccaacagcta gcgatactaa gtgtgtggta
     1801 ggacttcacg acaatactaa cgccgaattc gagtttcggg caaccacact ggaacggatc
     1861 gagaaaagct tcgtgattac tgatccgaga ctgcctgaca acccaatcat ttttgtgagc
     1921 gattccttcc tgcagctgac agaatattct cgggaagaga tcctggggcg caattgccgt
     1981 tttctgcagg gacccgagac agaccgtgcc actgttcgga aaatcagaga tgctattgac
     2041 aaccagactg aagtgaccgt tcagctgatc aattatacca agagcggcaa gaagttctgg
     2101 aacgtgttcc acctgcagcc gatgcgcgat tataagggcg acgtccagta cttcattggc
     2161 gtgcagctgg atggcaccga acgtcttcat ggcgccgctg agcgtgaggc ggtctgcctg
     2221 gtcaaaaaga cagcctttca gattgctgcg gccgctgata gagactatga tacagtcaac
     2281 ccagaagaga gtgccttgag agctcttgtt gcttcagcaa aaagtcacat tgaaatatct
     2341 cagcaggatt tgaacgctac atgtccaccc ttgcctaggt atgacattag attgtacgat
     2401 gccttggctg ctaaaatggc tgctggggtt aaggtgagaa ttgtggttag cgacccagcc
     2461 aacaggggcg cagtgggcag tgtgggttac agccaaataa aatcacttag tgaaatatca
     2521 gacactttga gaaacaggct tgcaaatatc acaggcagtc aacaagctgc caaaactgca
     2581 atgtgtagca atttgcagct tgcaaccttt agatcctccc ccaacgacaa atgggcggac
     2641 gggcatcctt atgcccaaca ccataagttg gtttccgtcg actcatccac attttatatc
     2701 ggttcaaaaa atttgtaccc atcctggctg caagattttg gctacatagt ggaatcccca
     2761 gaggctgcca agcaactgga tgccaaattg cttgaccctc agtggaagta ctcacaggag
     2821 actgcaacag tggattacgc aagaggaatc tgcaacgctg gtaccggagg cggttcaggc
     2881 ggaggtggca gcggcgtgag caagggcgag gaggataaca tggccatcat caaggagttc
     2941 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
     3001 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
     3061 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
     3121 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
     3181 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
     3241 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     3301 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     3361 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     3421 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     3481 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     3541 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     3601 gctcgaggcg cagcagcggg agcaggagga accggtggct cggctcgaga gggctggctc
     3661 ttcaaatgga ccaattatat caaaggctac cagcggcgat ggttcgtgct gagcaacggg
     3721 ctcctgagct actacagatc aaaggcagag atgagacata cctgccgtgg taccatcaac
     3781 ctcgccacag ccaacatcac cgtggaggac tcctgcaact tcatcatttc caatgggggt
     3841 gctcagacct accatctgaa agctagttca gaagttgagc ggcagcgctg ggtgacggcc
     3901 ctggaactgg ccaaggccaa agctgtgaag atgctggcat gagggcccta ttctatagtg
     3961 tcacctaaat gctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     4021 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     4081 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     4141 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     4201 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     4261 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4321 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4381 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     4441 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4501 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4561 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4621 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4681 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4741 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4801 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4861 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4921 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     4981 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     5041 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     5101 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     5161 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     5221 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5281 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5341 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5401 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5461 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5521 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5581 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     5641 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5701 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5761 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5821 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5881 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     5941 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     6001 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     6061 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     6121 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     6181 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     6241 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6301 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6361 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6421 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6481 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6541 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6601 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6661 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6721 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6781 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6841 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6901 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6961 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     7021 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7081 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7141 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7201 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7261 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7321 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7381 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7441 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7501 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7561 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7621 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7681 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7741 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7801 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7861 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7921 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7981 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     8041 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     8101 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8161 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8221 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8281 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8341 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
