LOCUS       sequence-474132-        5771 bp    DNA     circular SYN 30-MAY-2025
DEFINITION  Mammalian expression of recombinant BMPS WT.
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 May 30, 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 WT-6xHis
FEATURES             Location/Qualifiers
     source          1..5771
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     29..49
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     75..99
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        309..327
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     309..326
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     340..359
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1498..1515
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      1552..1561
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1565..1699
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1602..1621)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1656..1675
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        1768..2125
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1768..1959
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1791..1811)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1976..2111
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2145..2162)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2145..2161)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2154..2176)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2374..2829
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2461..2480)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2671..2692
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2846..2865)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2965..2987
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3025..3043)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3111..3215
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3216..3284,3285..4076)
                     /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(3434..3453)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4247..4835
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4736..4755
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4989..5006
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5123..5144
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5159..5164,5165..5182,5183..5189)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5197..5213
                     /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     5202..5224
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5221..5237
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5221..5237
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        5266..5645
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(5646..5771,1..78)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
       61 gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc
      121 tgttttgacc tccatagaag acaccgggac cgatccagcc tccgcggccg ggaacggtgc
      181 attggaacgc ggattccccg tgccaagagt gacgtaagta ccgcctatag agtctatagg
      241 cccaccccct tggcttcgtt agaacgcggc tacaattaat acataacctt atgtatcata
      301 cacatacgat ttaggtgaca ctatagaata acatccactt tgcctttctc tccacaggtg
      361 tccactccca ggtccaactg cacctcggtt ctatcgattg aattcaccat ggcccaggag
      421 gtcgacacag cacagggagc agagatgaga cggggcgccg gggccgccag aggacgggcc
      481 tcctggtgtt gggcactcgc cctcctgtgg ctggctgtcg tgcctgggtg gagcagagtt
      541 tcaggcatcc catctcgccg acactggcca gttccttata agagatttga cttcagacca
      601 aagcccgacc cgtattgcca ggccaagtac acattttgcc ctactgggtc acccataccc
      661 gtcatggagg gagacgatga catcgaagtg tttagattgc aagcccccgt gtgggagttc
      721 aagtacggtg acctgcttgg ccatctgaaa atcatgcatg acgcaatagg tttccggtct
      781 accctcacag gaaagaatta caccatggag tggtatgaac tcttccagct gggcaattgc
      841 acattcccac atctgagacc tgagatggat gcaccattct ggtgtaacca gggcgctgcc
      901 tgcttcttcg aagggatcga cgacgtgcat tggaaagaga atggcaccct ggtacaggtc
      961 gcgacaatta gcggcaatat gtttaaccag atggccaaat gggtgaagca ggacaacgag
     1021 acaggcatat actacgaaac atggaacgtg aaggcatcac ctgagaaggg cgctgaaact
     1081 tggttcgaca gttatgattg cagcaaattt gtcctgagaa cttttaataa gctggcagag
     1141 ttcggagccg aattcaaaaa tattgaaacc aattacacca ggatctttct ttattccggc
     1201 gagcccactt acctgggcaa cgaaacctca gtgttcggtc ctaccgggaa taagacgctc
     1261 ggtttggcca tcaaacgctt ctactaccct ttcaagccac atttgcccac caaggagttc
     1321 ctcctgagtc tcctgcagat ttttgatgct gtgatcgtcc ataagcagtt ttacttgttt
     1381 tacaatttcg aatattggtt cctccccatg aaattccctt tcatcaagat cacatacgag
     1441 gagatccccc tccccattag gaacaagact ctgtcaggcc tcgggggcgg cggaagccac
     1501 catcaccacc atcactaagc ggccgctaag taagtaagga tccccagctt ggccgccatg
     1561 gcccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     1621 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     1681 atcttatcat gtctggatcg ggaattaatt cggcgcagca ccatggcctg aaataacctc
     1741 tgaaagagga acttggttag gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg
     1801 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     1861 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     1921 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     1981 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     2041 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     2101 ttttggaggc ctaggctttt gcaaaaagct gttaacagct tggcactggc cgtcgtttta
     2161 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     2221 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     2281 cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     2341 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     2401 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     2461 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     2521 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     2581 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     2641 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     2701 tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     2761 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     2821 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     2881 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     2941 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     3001 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     3061 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3121 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3181 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     3241 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     3301 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     3361 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     3421 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     3481 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     3541 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3601 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3661 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3721 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     3781 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     3841 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     3901 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     3961 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4021 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     4081 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4141 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4201 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4261 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4321 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4381 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4441 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4501 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4561 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4621 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4681 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4741 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4801 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4861 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     4921 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     4981 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     5041 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     5101 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     5161 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     5221 caggaaacag ctatgacatg attacgaatt aattcgagct cgcccgacat tgattattga
     5281 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     5341 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     5401 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     5461 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     5521 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     5581 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     5641 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     5701 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     5761 gggactttcc a
//
