LOCUS       sequence-433905-        8150 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ER-targeted LOVPLD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8150)
  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 8150)
  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-C1-mCherry-LOVPLD
FEATURES             Location/Qualifiers
     source          1..8150
                     /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"
     CDS             1030..1734
                     /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(1172..1190)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1440..1459
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        complement(3703..3721)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3704..3721)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3741..3758)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3747..3971
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4017..4445
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4104..4123)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4314..4335
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4454..4474)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4459..4788
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4639..4774
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4701..4720
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4855..5649
                     /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(4909..4928)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5519..5538
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5823..5944
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5860..5879)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5914..5933
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5993..6009)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5993..6009)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6006..6028)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6017..6033
                     /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(6041..6047,6048..6065,6066..6071))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6086..6107
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6224..6241)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6395..6983)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6475..6494)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7154..7945,7946..8014))
                     /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     7777..7796
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8015..8119)
                     /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 atgatggacc ctgtggtcgt gctggggctg tgcctgagct gtctgctgct gctgagcctg
      961 tggaagcagt cctacggagg cgggggatca accggttcca gtggaggcgg tagctcaggt
     1021 tcgggtaccg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
     1081 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     1141 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
     1201 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
     1261 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
     1321 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
     1381 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     1441 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
     1501 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
     1561 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     1621 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
     1681 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagggatcc
     1741 gcagattcag ctacaccaca tttggacgcc gttgagcaaa cattgaggca agtgtcccca
     1801 ggcctggagg gcgatgtatg ggaaagaact agtggaaata aattggacgg aagtgccgct
     1861 gatcccagtg attggttgtt gcaaacccca ggatgttggg gcgatgatag atgcgttgac
     1921 agagttggga caaaaaggtt gctggctaag atgactgaaa atatagggaa tgccaccaga
     1981 accgtagata ttagtacact tgcacctttc ccaaacggcg cattccaaga cgctattgtc
     2041 gctggtttga aggagtccgc agctagaggc aataagttga aagttaggat actggttggg
     2101 gctgcacccg tctaccacat gaatgtaata ccttccaagt atagggacga attgactgca
     2161 aaacttggca aagccgcaga aaacataact ttgaacgtgg ctagtatgac tacatcaaaa
     2221 acagctttct cctggaacca tagcaaaatc ttggttgtag acgggcaaag cgcccttact
     2281 ggtggaataa atagctggaa ggatgattac ttggatacta ctcatccagt gtctgatgtt
     2341 gatcttgcac tgaccggacc agctgcaggc tctgccggta gataccttga cactttgtgg
     2401 acatggacct gccaaaacaa gagtaacatc gcttccgtct ggtttgcagc ttccggcaac
     2461 gcagggtgca tggccactat gcataaagat acaaacccca aggcatcacc tgccaccggg
     2521 aatgtcccaa taatagccgt tggagggttg ggcgttggca tcaaagatgt ggaccccaag
     2581 tccactttca ggcccgatct gccaacagct agcgatacta agtgtgtggt aggacttcac
     2641 gacaatacta acgccgaatt cgagtttcgg gcaaccacac tggaacggat cgagaaaagc
     2701 ttcgtgatta ctgatccgag actgcctgac aacccaatca tttttgtgag cgattccttc
     2761 ctgcagctga cagaatattc tcgggaagag atcctggggc gcaattgccg ttttctgcag
     2821 ggacccgaga cagaccgtgc cactgttcgg aaaatcagag atgctattga caaccagact
     2881 gaagtgaccg ttcagctgat caattatacc aagagcggca agaagttctg gaacgtgttc
     2941 cacctgcagc cgatgcgcga ttataagggc gacgtccagt acttcattgg cgtgcagctg
     3001 gatggcaccg aacgtcttca tggcgccgct gagcgtgagg cggtctgcct ggtcaaaaag
     3061 acagcctttc agattgctgc ggccgctgat agagactatg atacagtcaa cccagaagag
     3121 agtgccttga gagctcttgt tgcttcagca aaaagtcaca ttgaaatatc tcagcaggat
     3181 ttgaacgcta catgtccacc cttgcctagg tatgacatta gattgtacga tgccttggct
     3241 gctaaaatgg ctgctggggt taaggtgaga attgtggtta gcgacccagc caacaggggc
     3301 gcagtgggca gtgtgggtta cagccaaata aaatcactta gtgaaatatc agacactttg
     3361 agaaacaggc ttgcaaatat cacaggcagt caacaagctg ccaaaactgc aatgtgtagc
     3421 aatttgcagc ttgcaacctt tagatcctcc cccaacgaca aatgggcgga cgggcatcct
     3481 tatgcccaac accataagtt ggtttccgtc gactcatcca cattttatat cggttcaaaa
     3541 aatttgtacc catcctggct gcaagatttt ggctacatag tggaatcccc agaggctgcc
     3601 aagcaactgg atgccaaatt gcttgaccct cagtggaagt actcacagga gactgcaaca
     3661 gtggattacg caagaggaat ctgcaacgct taagggccct attctatagt gtcacctaaa
     3721 tgctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     3781 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3841 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3901 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     3961 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     4021 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     4081 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     4141 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     4201 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     4261 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     4321 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     4381 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     4441 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4501 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4561 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4621 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4681 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     4741 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     4801 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4861 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4921 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4981 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     5041 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     5101 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     5161 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     5221 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     5281 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     5341 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     5401 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5461 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5521 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5581 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5641 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5701 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5761 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5821 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5881 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5941 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     6001 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     6061 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     6121 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     6181 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     6241 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     6301 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     6361 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     6421 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6481 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6541 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6601 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6661 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6721 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6781 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6841 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6901 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6961 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     7021 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     7081 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     7141 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     7201 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     7261 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     7321 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     7381 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     7441 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     7501 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7561 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7621 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7681 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7741 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     7801 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7861 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7921 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7981 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     8041 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     8101 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
