LOCUS       sequence-474135-        5771 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of recombinant BMPS C231S.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5771)
  AUTHORS   Medoh UN, Hims A, Chen JY, Ghoochani A, Nyame K, Dong W, 
            Abu-Remaileh M
  TITLE     The Batten disease gene product CLN5 is the lysosomal 
            bis(monoacylglycero)phosphate synthase.
  JOURNAL   Science. 2023 Sep 15;381(6663):1182-1189. doi: 
            10.1126/science.adg9288.
  PUBMED    37708259
REFERENCE   2  (bases 1 to 5771)
  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: "10.1126/science"; 
            journalName: "Science"; date: "2023-09-15- 15"; volume: "381"; 
            issue: "6663"; pages: "1182-1189"
COMMENT     Sequence Label: pRK5 BMPS C231S-6xHis
FEATURES             Location/Qualifiers
     source          1..5771
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             918..935
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      972..981
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    985..1119
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1022..1041)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1076..1095
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        1188..1545
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1188..1379
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1211..1231)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1396..1531
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1565..1582)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1565..1581)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1574..1596)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1794..2249
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1881..1900)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2091..2112
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2266..2285)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2385..2407
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2445..2463)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2531..2635
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2636..2704,2705..3496)
                     /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(2854..2873)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3667..4255
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4156..4175
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4409..4426
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4543..4564
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4579..4584,4585..4602,4603..4609)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4617..4633
                     /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     4622..4644
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4641..4657
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4641..4657
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        4686..5065
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5066..5269
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5220..5240
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5266..5290
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        5500..5518
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5500..5517
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     5531..5550
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 agagatttga cttcagacca aagcccgacc cgtattgcca ggccaagtac acattttgcc
       61 ctactgggtc acccataccc gtcatggagg gagacgatga catcgaagtg tttagattgc
      121 aagcccccgt gtgggagttc aagtacggtg acctgcttgg ccatctgaaa atcatgcatg
      181 acgcaatagg tttccggtct accctcacag gaaagaatta caccatggag tggtatgaac
      241 tcttccagct gggcaattgc acattcccac atctgagacc tgagatggat gcaccattct
      301 ggtgtaacca gggcgctgcc tgcttcttcg aagggatcga cgacgtgcat tggaaagaga
      361 atggcaccct ggtacaggtc gcgacaatta gcggcaatat gtttaaccag atggccaaat
      421 gggtgaagca ggacaacgag acaggcatat actacgaaac atggaacgtg aaggcatcac
      481 ctgagaaggg cgctgaaact tggttcgaca gttatgatag tagcaaattt gtcctgagaa
      541 cttttaataa gctggcagag ttcggagccg aattcaaaaa tattgaaacc aattacacca
      601 ggatctttct ttattccggc gagcccactt acctgggcaa cgaaacctca gtgttcggtc
      661 ctaccgggaa taagacgctc ggtttggcca tcaaacgctt ctactaccct ttcaagccac
      721 atttgcccac caaggagttc ctcctgagtc tcctgcagat ttttgatgct gtgatcgtcc
      781 ataagcagtt ttacttgttt tacaatttcg aatattggtt cctccccatg aaattccctt
      841 tcatcaagat cacatacgag gagatccccc tccccattag gaacaagact ctgtcaggcc
      901 tcgggggcgg cggaagccac catcaccacc atcactaagc ggccgctaag taagtaagga
      961 tccccagctt ggccgccatg gcccaacttg tttattgcag cttataatgg ttacaaataa
     1021 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     1081 ttgtccaaac tcatcaatgt atcttatcat gtctggatcg ggaattaatt cggcgcagca
     1141 ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg aggcggaaag
     1201 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     1261 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     1321 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     1381 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     1441 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     1501 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct gttaacagct
     1561 tggcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     1621 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     1681 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc
     1741 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     1801 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     1861 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     1921 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     1981 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     2041 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     2101 gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat
     2161 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     2221 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     2281 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     2341 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     2401 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     2461 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     2521 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2581 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     2641 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     2701 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     2761 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     2821 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     2881 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     2941 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3001 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     3061 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     3121 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     3181 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     3241 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3301 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3361 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3421 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3481 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     3541 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     3601 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3661 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3721 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3781 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3841 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3901 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3961 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4021 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4081 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4141 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4201 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4261 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4321 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     4381 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     4441 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     4501 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     4561 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     4621 gagcggataa caatttcaca caggaaacag ctatgacatg attacgaatt aattcgagct
     4681 cgcccgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     4741 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     4801 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     4861 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     4921 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     4981 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     5041 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     5101 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     5161 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     5221 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa
     5281 ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccggga
     5341 ccgatccagc ctccgcggcc gggaacggtg cattggaacg cggattcccc gtgccaagag
     5401 tgacgtaagt accgcctata gagtctatag gcccaccccc ttggcttcgt tagaacgcgg
     5461 ctacaattaa tacataacct tatgtatcat acacatacga tttaggtgac actatagaat
     5521 aacatccact ttgcctttct ctccacaggt gtccactccc aggtccaact gcacctcggt
     5581 tctatcgatt gaattcacca tggcccagga ggtcgacaca gcacagggag cagagatgag
     5641 acggggcgcc ggggccgcca gaggacgggc ctcctggtgt tgggcactcg ccctcctgtg
     5701 gctggctgtc gtgcctgggt ggagcagagt ttcaggcatc ccatctcgcc gacactggcc
     5761 agttccttat a
//
