LOCUS       sequence-433902-        8222 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mitochondria-targeted LOVPLD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8222)
  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 8222)
  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-AKAP1-mCherry-LOVPLD
FEATURES             Location/Qualifiers
     source          1..8222
                     /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             1102..1806
                     /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(1244..1262)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1512..1531
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        complement(3775..3793)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3776..3793)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3813..3830)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3819..4043
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4089..4517
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4176..4195)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4386..4407
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4526..4546)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4531..4860
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4711..4846
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4773..4792
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4927..5721
                     /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(4981..5000)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5591..5610
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5895..6016
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5932..5951)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5986..6005
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6065..6081)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6065..6081)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6078..6100)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6089..6105
                     /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(6113..6119,6120..6137,6138..6143))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6158..6179
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6296..6313)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6467..7055)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6547..6566)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7226..8017,8018..8086))
                     /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     7849..7868
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8087..8191)
                     /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 atgatggcca tccagctgcg cagcctgttc cccctggccc tgcccggcct gctggccctg
      961 ctgggctggt ggtggttctt cagccgcaag aaggacctgg agctgaagct gcgcatcctg
     1021 cagagcaccg tgccccgcgc ccgcgaccct cccgtggcca ccaccggttc cagtggaggc
     1081 ggtagctcag gttcgggtac cgtgagcaag ggcgaggagg ataacatggc catcatcaag
     1141 gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc
     1201 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
     1261 aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc
     1321 aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag
     1381 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag
     1441 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc
     1501 ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg
     1561 atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac
     1621 ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg
     1681 cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc
     1741 atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg
     1801 tacaagggat ccgcagattc agctacacca catttggacg ccgttgagca aacattgagg
     1861 caagtgtccc caggcctgga gggcgatgta tgggaaagaa ctagtggaaa taaattggac
     1921 ggaagtgccg ctgatcccag tgattggttg ttgcaaaccc caggatgttg gggcgatgat
     1981 agatgcgttg acagagttgg gacaaaaagg ttgctggcta agatgactga aaatataggg
     2041 aatgccacca gaaccgtaga tattagtaca cttgcacctt tcccaaacgg cgcattccaa
     2101 gacgctattg tcgctggttt gaaggagtcc gcagctagag gcaataagtt gaaagttagg
     2161 atactggttg gggctgcacc cgtctaccac atgaatgtaa taccttccaa gtatagggac
     2221 gaattgactg caaaacttgg caaagccgca gaaaacataa ctttgaacgt ggctagtatg
     2281 actacatcaa aaacagcttt ctcctggaac catagcaaaa tcttggttgt agacgggcaa
     2341 agcgccctta ctggtggaat aaatagctgg aaggatgatt acttggatac tactcatcca
     2401 gtgtctgatg ttgatcttgc actgaccgga ccagctgcag gctctgccgg tagatacctt
     2461 gacactttgt ggacatggac ctgccaaaac aagagtaaca tcgcttccgt ctggtttgca
     2521 gcttccggca acgcagggtg catggccact atgcataaag atacaaaccc caaggcatca
     2581 cctgccaccg ggaatgtccc aataatagcc gttggagggt tgggcgttgg catcaaagat
     2641 gtggacccca agtccacttt caggcccgat ctgccaacag ctagcgatac taagtgtgtg
     2701 gtaggacttc acgacaatac taacgccgaa ttcgagtttc gggcaaccac actggaacgg
     2761 atcgagaaaa gcttcgtgat tactgatccg agactgcctg acaacccaat catttttgtg
     2821 agcgattcct tcctgcagct gacagaatat tctcgggaag agatcctggg gcgcaattgc
     2881 cgttttctgc agggacccga gacagaccgt gccactgttc ggaaaatcag agatgctatt
     2941 gacaaccaga ctgaagtgac cgttcagctg atcaattata ccaagagcgg caagaagttc
     3001 tggaacgtgt tccacctgca gccgatgcgc gattataagg gcgacgtcca gtacttcatt
     3061 ggcgtgcagc tggatggcac cgaacgtctt catggcgccg ctgagcgtga ggcggtctgc
     3121 ctggtcaaaa agacagcctt tcagattgct gcggccgctg atagagacta tgatacagtc
     3181 aacccagaag agagtgcctt gagagctctt gttgcttcag caaaaagtca cattgaaata
     3241 tctcagcagg atttgaacgc tacatgtcca cccttgccta ggtatgacat tagattgtac
     3301 gatgccttgg ctgctaaaat ggctgctggg gttaaggtga gaattgtggt tagcgaccca
     3361 gccaacaggg gcgcagtggg cagtgtgggt tacagccaaa taaaatcact tagtgaaata
     3421 tcagacactt tgagaaacag gcttgcaaat atcacaggca gtcaacaagc tgccaaaact
     3481 gcaatgtgta gcaatttgca gcttgcaacc tttagatcct cccccaacga caaatgggcg
     3541 gacgggcatc cttatgccca acaccataag ttggtttccg tcgactcatc cacattttat
     3601 atcggttcaa aaaatttgta cccatcctgg ctgcaagatt ttggctacat agtggaatcc
     3661 ccagaggctg ccaagcaact ggatgccaaa ttgcttgacc ctcagtggaa gtactcacag
     3721 gagactgcaa cagtggatta cgcaagagga atctgcaacg cttaagggcc ctattctata
     3781 gtgtcaccta aatgctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
     3841 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3901 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     3961 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     4021 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     4081 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4141 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4201 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4261 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     4321 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4381 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4441 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     4501 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4561 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4621 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4681 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4741 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     4801 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4861 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     4921 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4981 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     5041 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     5101 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     5161 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     5221 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     5281 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     5341 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     5401 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     5461 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5521 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5581 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5641 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5701 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5761 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5821 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5881 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5941 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     6001 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     6061 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     6121 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     6181 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     6241 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     6301 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     6361 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     6421 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6481 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6541 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6601 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6661 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6721 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6781 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6841 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6901 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6961 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     7021 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     7081 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     7141 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     7201 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     7261 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     7321 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     7381 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     7441 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7501 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7561 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7621 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7681 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7741 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7801 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7861 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7921 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7981 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     8041 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     8101 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     8161 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     8221 tc
//
