LOCUS       sequence-454650-        5771 bp    DNA     circular SYN 14-APR-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 Apr 14, 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 HK318/9EE-6xHis
FEATURES             Location/Qualifiers
     source          1..5771
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    6..140
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(43..62)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     97..116
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        209..566
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        209..400
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(232..252)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      417..552
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(586..603)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(586..602)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(595..617)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      815..1270
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(902..921)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1112..1133
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1287..1306)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1406..1428
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1466..1484)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1552..1656
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1657..1725,1726..2517)
                     /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(1875..1894)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2688..3276
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3177..3196
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3430..3447
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3564..3585
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3600..3605,3606..3623,3624..3630)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3638..3654
                     /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     3643..3665
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3662..3678
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3662..3678
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        3707..4086
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4087..4290
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4241..4261
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4287..4311
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        4521..4539
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4521..4538
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     4552..4571
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             5710..5727
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      join(5764..5771,1..2)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ggcccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
       61 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
      121 tatcttatca tgtctggatc gggaattaat tcggcgcagc accatggcct gaaataacct
      181 ctgaaagagg aacttggtta ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt
      241 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
      301 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
      361 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
      421 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
      481 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
      541 tttttggagg cctaggcttt tgcaaaaagc tgttaacagc ttggcactgg ccgtcgtttt
      601 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
      661 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
      721 gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
      781 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
      841 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
      901 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
      961 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     1021 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     1081 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     1141 ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     1201 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     1261 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     1321 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     1381 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     1441 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     1501 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     1561 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     1621 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     1681 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     1741 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     1801 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     1861 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     1921 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     1981 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     2041 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     2101 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     2161 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     2221 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     2281 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     2341 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     2401 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     2461 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     2521 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     2581 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     2641 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     2701 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     2761 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     2821 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     2881 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     2941 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3001 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3061 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3121 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3181 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3241 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     3301 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     3361 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     3421 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     3481 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     3541 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     3601 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     3661 acaggaaaca gctatgacat gattacgaat taattcgagc tcgcccgaca ttgattattg
     3721 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     3781 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     3841 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     3901 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     3961 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     4021 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     4081 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
     4141 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     4201 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
     4261 gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga
     4321 cgccatccac gctgttttga cctccataga agacaccggg accgatccag cctccgcggc
     4381 cgggaacggt gcattggaac gcggattccc cgtgccaaga gtgacgtaag taccgcctat
     4441 agagtctata ggcccacccc cttggcttcg ttagaacgcg gctacaatta atacataacc
     4501 ttatgtatca tacacatacg atttaggtga cactatagaa taacatccac tttgcctttc
     4561 tctccacagg tgtccactcc caggtccaac tgcacctcgg ttctatctat tgaattcacc
     4621 atggcccagg aggtcgacac agcacaggga gcagagatga gacggggcgc cggggccgcc
     4681 agaggacggg cctcctggtg ttgggcactc gccctcctgt ggctggctgt cgtgcctggg
     4741 tggagcagag tttcaggcat cccatctcgc cgacactggc cagttcctta taagagattt
     4801 gacttcagac caaagcccga cccgtattgc caggccaagt acacattttg ccctactggg
     4861 tcacccatac ccgtcatgga gggagacgat gacatcgaag tgtttagatt gcaagccccc
     4921 gtgtgggagt tcaagtacgg tgacctgctt ggccatctga aaatcatgca tgacgcaata
     4981 ggtttccggt ctaccctcac aggaaagaat tacaccatgg agtggtatga actcttccag
     5041 ctgggcaatt gcacattccc acatctgaga cctgagatgg atgcaccatt ctggtgtaac
     5101 cagggcgctg cctgcttctt cgaagggatc gacgacgtgc attggaaaga gaatggcacc
     5161 ctggtacagg tcgcgacaat tagcggcaat atgtttaacc agatggccaa atgggtgaag
     5221 caggacaacg agacaggcat atactacgaa acatggaacg tgaaggcatc acctgagaag
     5281 ggcgctgaaa cttggttcga cagttatgat tgcagcaaat ttgtcctgag aacttttaat
     5341 aagctggcag agttcggagc cgaattcaaa aatattgaaa ccaattacac caggatcttt
     5401 ctttattccg gcgagcccac ttacctgggc aacgaaacct cagtgttcgg tcctaccggg
     5461 aataagacgc tcggtttggc catcaaacgc ttctactacc ctttcaagcc acatttgccc
     5521 accaaggagt tcctcctgag tctcctgcag atttttgatg ctgtgatcgt cgaagaacag
     5581 ttttacttgt tttacaattt cgaatattgg ttcctcccca tgaaattccc tttcatcaag
     5641 atcacatacg aggagatccc cctccccatt aggaacaaga ctctgtcagg cctcgggggc
     5701 ggcggaagcc accatcacca ccatcactaa gcggccgcta agtaagtaag gatccccagc
     5761 ttggccgcca t
//
