LOCUS       sequence-433901-        8093 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PM-targeted catalytically dead LOVPLD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8093)
  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 8093)
  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-lyn10-mCherry-dLOVPLD
FEATURES             Location/Qualifiers
     source          1..8093
                     /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             973..1677
                     /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(1115..1133)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1383..1402
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        complement(3646..3664)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3647..3664)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3684..3701)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3690..3914
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3960..4388
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4047..4066)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4257..4278
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4397..4417)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4402..4731
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4582..4717
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4644..4663
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4798..5592
                     /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(4852..4871)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5462..5481
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5766..5887
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5803..5822)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5857..5876
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5936..5952)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5936..5952)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5949..5971)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5960..5976
                     /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(5984..5990,5991..6008,6009..6014))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6029..6050
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6167..6184)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6338..6926)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6418..6437)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7097..7888,7889..7957))
                     /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     7720..7739
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7958..8062)
                     /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 atgggatgta ttaaatcaaa aaggaaagac gggaccggtt ccagtggagg cggtagctca
      961 ggttcgggta ccgtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg
     1021 cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
     1081 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc
     1141 cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac
     1201 gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag
     1261 tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc
     1321 ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac
     1381 ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc
     1441 gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac
     1501 tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc
     1561 tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa
     1621 cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaaggga
     1681 tccgcagatt cagctacacc acatttggac gccgttgagc aaacattgag gcaagtgtcc
     1741 ccaggcctgg agggcgatgt atgggaaaga actagtggaa ataaattgga cggaagtgcc
     1801 gctgatccca gtgattggtt gttgcaaacc ccaggatgtt ggggcgatga tagatgcgtt
     1861 gacagagttg ggacaaaaag gttgctggct aagatgactg aaaatatagg gaatgccacc
     1921 agaaccgtag atattagtac acttgcacct ttcccaaacg gcgcattcca agacgctatt
     1981 gtcgctggtt tgaaggagtc cgcagctaga ggcaataagt tgaaagttag gatactggtt
     2041 ggggctgcac ccgtctacca catgaatgta ataccttcca agtataggga cgaattgact
     2101 gcaaaacttg gcaaagccgc agaaaacata actttgaacg tggctagtat gactacatca
     2161 aaaacagctt tctcctggaa cgctagcaaa atcttggttg tagacgggca aagcgccctt
     2221 actggtggaa taaatagctg gaaggatgat tacttggata ctactcatcc agtgtctgat
     2281 gttgatcttg cactgaccgg accagctgca ggctctgccg gtagatacct tgacactttg
     2341 tggacatgga cctgccaaaa caagagtaac atcgcttccg tctggtttgc agcttccggc
     2401 aacgcagggt gcatggccac tatgcataaa gatacaaacc ccaaggcatc acctgccacc
     2461 gggaatgtcc caataatagc cgttggaggg ttgggcgttg gcatcaaaga tgtggacccc
     2521 aagtccactt tcaggcccga tctgccaaca gctagcgata ctaagtgtgt ggtaggactt
     2581 cacgacaata ctaacgccga attcgagttt cgggcaacca cactggaacg gatcgagaaa
     2641 agcttcgtga ttactgatcc gagactgcct gacaacccaa tcatttttgt gagcgattcc
     2701 ttcctgcagc tgacagaata ttctcgggaa gagatcctgg ggcgcaattg ccgttttctg
     2761 cagggacccg agacagaccg tgccactgtt cggaaaatca gagatgctat tgacaaccag
     2821 actgaagtga ccgttcagct gatcaattat accaagagcg gcaagaagtt ctggaacgtg
     2881 ttccacctgc agccgatgcg cgattataag ggcgacgtcc agtacttcat tggcgtgcag
     2941 ctggatggca ccgaacgtct tcatggcgcc gctgagcgtg aggcggtctg cctggtcaaa
     3001 aagacagcct ttcagattgc tgcggccgct gatagagact atgatacagt caacccagaa
     3061 gagagtgcct tgagagctct tgttgcttca gcaaaaagtc acattgaaat atctcagcag
     3121 gatttgaacg ctacatgtcc acccttgcct aggtatgaca ttagattgta cgatgccttg
     3181 gctgctaaaa tggctgctgg ggttaaggtg agaattgtgg ttagcgaccc agccaacagg
     3241 ggcgcagtgg gcagtgtggg ttacagccaa ataaaatcac ttagtgaaat atcagacact
     3301 ttgagaaaca ggcttgcaaa tatcacaggc agtcaacaag ctgccaaaac tgcaatgtgt
     3361 agcaatttgc agcttgcaac ctttagatcc tcccccaacg acaaatgggc ggacgggcat
     3421 ccttatgccc aacaccataa gttggtttcc gtcgactcat ccacatttta tatcggttca
     3481 aaaaatttgt acccatcctg gctgcaagat tttggctaca tagtggaatc cccagaggct
     3541 gccaagcaac tggatgccaa attgcttgac cctcagtgga agtactcaca ggagactgca
     3601 acagtggatt acgcaagagg aatctgcaac gcttaagggc cctattctat agtgtcacct
     3661 aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     3721 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     3781 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     3841 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     3901 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     3961 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4021 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4081 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4141 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4201 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4261 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4321 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4381 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4441 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4501 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4561 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4621 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     4681 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4741 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4801 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4861 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4921 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     4981 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     5041 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     5101 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     5161 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     5221 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     5281 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     5341 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     5401 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5461 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5521 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5581 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     5641 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     5701 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     5761 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5821 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5881 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     5941 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     6001 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     6061 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     6121 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6181 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6241 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6301 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6361 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6421 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6481 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6541 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6601 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6661 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6721 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6781 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6841 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6901 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6961 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7021 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7081 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7141 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7201 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7261 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7321 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7381 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7441 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7501 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7561 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7621 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7681 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7741 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7801 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7861 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7921 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7981 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     8041 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
