LOCUS       sequence-424660-        5933 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses PLA2G15-6xHis in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5933)
  AUTHORS   Nyame K, Hims A, Aburous A, Laqtom NN, Dong W, Medoh UN, Heiby JC, 
            Xiong J, Ori A, Abu-Remaileh M
  TITLE     Glycerophosphodiesters inhibit lysosomal phospholipid catabolism in 
            Batten disease.
  JOURNAL   Mol Cell. 2024 Apr 4;84(7):1354-1364.e9. doi: 
            10.1016/j.molcel.2024.02.006. Epub 2024 Mar 5.
  PUBMED    38447580
REFERENCE   2  (bases 1 to 5933)
  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: 
            "10.1016/j.molcel.2024.02.006"; journalName: "Mol Cell"; date: 
            "2024-04-4- 4"; volume: "84"; issue: "7"; pages: "1354-1364.e9"
COMMENT     Sequence Label: pRK5-PLA2G15-6xHis WT
FEATURES             Location/Qualifiers
     source          1..5933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     828..845
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     859..878
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             2179..2196
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      2233..2242
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2246..2380
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2283..2302)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2337..2356
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2449..2806
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2449..2640
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2472..2492)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2657..2792
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2826..2843)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2826..2842)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2835..2857)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3055..3510
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3142..3161)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3352..3373
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3527..3546)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3646..3668
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3706..3724)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3792..3896
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3897..3965,3966..4757)
                     /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     complement(4115..4134)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4928..5516
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5417..5436
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5670..5687
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5804..5825
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5840..5845,5846..5863,5864..5870)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5878..5894
                     /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)."
     primer_bind     5883..5905
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5902..5918
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5902..5918
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      901 acctcggttc tatcgattga attcaccatg ggcctccacc tccgccccta ccgtgtgggg
      961 ctgctcccgg atggcctcct gttcctcttg ctgctgctaa tgctgctcgc ggacccagcg
     1021 ctcccggccg gacgtcaccc cccagtggtg ctggtccctg gtgatttggg taaccaactg
     1081 gaagccaagc tggacaagcc gacagtggtg cactacctct gctccaagaa gaccgaaagc
     1141 tacttcacaa tctggctgaa cctggaactg ctgctgcctg tcatcattga ctgctggatt
     1201 gacaatatca ggctggttta caacaaaaca tccagggcca cccagtttcc tgatggtgtg
     1261 gatgtacgtg tccctggctt tgggaagacc ttctcactgg agttcctgga ccccagcaaa
     1321 agcagcgtgg gttcctattt ccacaccatg gtggagagcc ttgtgggctg gggctacaca
     1381 cggggtgagg atgtccgagg ggctccctat gactggcgcc gagccccaaa tgaaaacggg
     1441 ccctacttcc tggccctccg cgagatgatc gaggagatgt accagctgta tgggggcccc
     1501 gtggtgctgg ttgcccacag tatgggcaac atgtacacgc tctactttct gcagcggcag
     1561 ccgcaggcct ggaaggacaa gtatatccgg gccttcgtgt cactgggtgc gccctggggg
     1621 ggcgtggcca agaccctgcg cgtcctggct tcaggagaca acaaccggat cccagtcatc
     1681 gggcccctga agatccggga gcagcagcgg tcagctgtct ccaccagctg gctgctgccc
     1741 tacaactaca catggtcacc tgagaaggtg ttcgtgcaga cacccacaat caactacaca
     1801 ctgcgggact accgcaagtt cttccaggac atcggctttg aagatggctg gctcatgcgg
     1861 caggacacag aagggctggt ggaagccacg atgccacctg gcgtgcagct gcactgcctc
     1921 tatggtactg gcgtccccac accagactcc ttctactatg agagcttccc tgaccgtgac
     1981 cctaaaatct gctttggtga cggcgatggt actgtgaact tgaagagtgc cctgcagtgc
     2041 caggcctggc agagccgcca ggagcaccaa gtgttgctgc aggagctgcc aggcagcgag
     2101 cacatcgaga tgctggccaa cgccaccacc ctggcctatc tgaaacgtgt gctccttggg
     2161 cccggaggtg gaggtagtca tcatcaccat caccactagg cggccgctaa gtaagtaagg
     2221 atccccagct tggccgccat ggcccaactt gtttattgca gcttataatg gttacaaata
     2281 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     2341 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc gggaattaat tcggcgcagc
     2401 accatggcct gaaataacct ctgaaagagg aacttggtta ggtaccttct gaggcggaaa
     2461 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     2521 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     2581 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     2641 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     2701 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     2761 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tgttaacagc
     2821 ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     2881 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     2941 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt
     3001 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     3061 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3121 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3181 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3241 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     3301 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3361 tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga
     3421 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3481 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     3541 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3601 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3661 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     3721 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     3781 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3841 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     3901 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3961 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     4021 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4081 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     4141 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4201 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     4261 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     4321 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     4381 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     4441 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     4501 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     4561 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     4621 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4681 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4741 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4801 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4861 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4921 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4981 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5041 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     5101 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5161 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5221 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5281 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5341 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5401 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5461 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5521 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5581 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     5641 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     5701 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     5761 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     5821 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     5881 tgagcggata acaatttcac acaggaaaca gctatgacat gattacgaat taa
//
