LOCUS       sequence-457149-        8118 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express N-terminal three tandem FLAG epitope tags, followed by an 
            enterokinase cleavage site and human DGKD gene deleted SAM domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8118)
  AUTHORS   Murakami C, Hoshino F, Sakai H, Hayashi Y, Yamashita A, Sakane F
  TITLE     Diacylglycerol kinase delta and sphingomyelin synthase-related 
            protein functionally interact via their sterile alpha motif domains.
  JOURNAL   J Biol Chem. 2020 Mar 6;295(10):2932-2947. doi: 
            10.1074/jbc.RA119.012369. Epub 2020 Jan 24.
  PUBMED    31980461
REFERENCE   2  (bases 1 to 8118)
  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: 
            "10.1074/jbc.RA119.012369"; journalName: "J Biol Chem"; date: 
            "2020-03-6- 6"; volume: "295"; issue: "10"; pages: "2932-2947"
COMMENT     Sequence Label: 3xFLAG-DGK-delta-2-Delta-SAMD
FEATURES             Location/Qualifiers
     source          1..8118
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    2579..3201
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2778..2797)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3225..3245)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3230..3559
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3410..3545
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3597..3616)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3598..3616)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3630..3646)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3630..3646)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3643..3665)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3654..3670
                     /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(3678..3684,3685..3702,3703..3708))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3723..3744
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3861..3878)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4032..4620)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4112..4131)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4791..5582,5583..5651))
                     /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     5414..5433
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5652..5756)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5783..6238
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5870..5889)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6080..6101
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6364..6386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6378..6395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6379..6395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        6556..6935
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6936..7139
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7090..7110
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             7169..7234
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
ORIGIN
        1 cctgtgcaaa gtgtcagtca tcccacccac ggctctcaac agcatcgact ccgatgggtt
       61 ctggaaggcc agctgtcctc cttcttgcac aagcccactg ttggtcttcg tcaattcaaa
      121 aagtggggac aaccagggtg tgaagttcct cagaagattc aaacagctac taaaccccgc
      181 ccaggtcttc gacctcatga acggaggccc acacctcggc ttacggttat tccagaagtt
      241 tgacacattc cggattctgg tttgtggcgg ggatggaagt gttggctggg tcctctccga
      301 aatcgacagc ctcaaccttc ataaacagtg tcagctggga gtgctgccgc tcggcacagg
      361 gaacgacttg gcccgagtac tgggctgggg ctcagcctgc gatgacgaca cccagctccc
      421 ccagatcttg gagaagttgg agagagccag caccaagatg ctggacaggt ggagcgtcat
      481 ggcatacgag gccaagctcc cccggcaggc ctcctcctct accgtcaccg aagacttcag
      541 cgaggattcc gaggtacagc agattctctt ctatgaagac tcggttgcag cccacctttc
      601 taaaatcctc acctcggacc agcactcggt ggtcatctcc tcggccaaag tgctctgtga
      661 gacggtgaag gacttcgtgg cacgggtggg gaaggcctat gagaagacga ccgagagctc
      721 ggaggagtca gaggtcatgg ccaagaagtg ctctgtcctg aaagagaagc tggattccct
      781 tctcaagacc ttggacgatg agtcccaggc ctcgtcctct ctgcccaacc cgccccccac
      841 cattgccgag gaggctgaag atggagatgg gtcgggcagc atctgcggtt ccaccggaga
      901 ccgcttggtg gcatcagctt gcccggcccg gccgcagata ttccggcctc gagaacagct
      961 catgctgaga gccaacagcc tgaagaaagc aattcgtcag atcatagaac acacagaaaa
     1021 agctgtcgat gagcagaatg cccagaccca ggagcaggag ggcttcgtcc tgggcctctc
     1081 tgagtcagag gagaagatgg accacagagt gtgcccacca ctgtcccaca gcgagagctt
     1141 cggggtcccc aaggggagga gccagcgcaa agtgtcgaaa tctccgtgtg aaaagctgat
     1201 cagcaaaggg agtctgtccc taggcagttc tgcttccctt ccgccccagc cgggaagccg
     1261 ggacggcctg cctgcgctca acaccaagat cctgtaccca aatgtccggg ctggaatgtc
     1321 tggttcctta cccggtggct cagtcatcag tcgcctgtta attaatgctg atcccttcaa
     1381 ctctgaacca gaaaccctag agtattacac ggagaaatgt gtcatgaaca actattttgg
     1441 cattggcctg gatgcgaaga tatccctgga ctttaacaac aagcgcgatg agcacccaga
     1501 gaagtgcagg agccgaacca agaacatgat gtggtatgga gttcttggaa ccaaagagtt
     1561 gctgcacaga acctacaaga acctggagca aaaggtcttg ctggagtgtg acgggcgacc
     1621 catcccactc cccagtcttc agggaattgc tgtccttaac attcccagct atgccggagg
     1681 aaccaacttc tgggggggta ccaaggaaga tgatactttc gcagctccat cattcgatga
     1741 caagattctg gaggtggtcg ccgtgttcgg cagcatgcag atggccgtct ctcgagtcat
     1801 caggctacag catcatcgga tcgcccagtg tcgcacggtg aagatctcca tccttgggga
     1861 tgagggcgtg cctgtgcagg tggacggaga ggcctgggtc cagccgccag ggtacattcg
     1921 gattgtccac aagaaccggg cacagacact gaccagagac agggcatttg agagcaccct
     1981 gaagtcctgg gaagacaagc agaagtgcga gctgccccgc cctccatcct gttccctgca
     2041 cccggagatg ctgtccgagg aggaggccac ccagatggac cagtttgggc aggcagcagg
     2101 ggtcctcatt cacagtatcc gagaaatagc tcagtctcac cgggacatgg agcaggaact
     2161 ggcccacgcc gtcaatgcca gctccaagtc catggaccgt gtgtatggca agcccagaac
     2221 cacagagggg ctcaactgca gcttcgtcct ggaaatggtg aataacttca gagctctgcg
     2281 cagtgagacg gagctgctgc tgtctgggaa gatggccctg cagctggatc cgcctcagaa
     2341 ggagcagctg gggagtgctc ttgccgagat ggaccgacag ctcaggaggc tggcagacac
     2401 cccgtggctc tgccagtccg cagagcccgg cgacgaagag agtgtgatgc tggatcttgc
     2461 caagcgcagt cgcagtggta aattccgcct cgtgaccaag tttaaaaagg agaaaaacaa
     2521 caagaacaaa gaagctcaca gtagcctggg agccccgtag tcgactctag aggatcccgg
     2581 gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt
     2641 gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt
     2701 ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac
     2761 ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct
     2821 cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt
     2881 gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg
     2941 tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg
     3001 gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat
     3061 tttaaaataa ctataccagc aggaggacgt ccagacacag cataggctac ctggccatgc
     3121 ccaaccggtg ggacatttga gttgcttgct tggcactgtc ctctcatgcg ttgggtccac
     3181 tcagtagatg cctgttgaat tgggtacgcg gccagcttgg ctgtggaatg tgtgtcagtt
     3241 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3301 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3361 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3421 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3481 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     3541 aggcctaggc ttttgcaaaa agctcctcga ggaactgaaa aaccagaaag ttaattccct
     3601 atagtgagtc gtattaaatt cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat
     3661 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     3721 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     3781 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3841 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3901 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3961 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4021 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4081 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4141 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4201 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4261 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4321 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4381 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4441 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4501 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4561 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4621 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4681 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4741 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4801 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4861 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4921 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4981 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5041 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5101 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5161 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5221 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5281 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5341 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5401 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5461 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5521 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     5581 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5641 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     5701 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5761 tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     5821 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     5881 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     5941 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     6001 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     6061 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     6121 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     6181 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc
     6241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     6301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     6361 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctgatctat acattgaatc
     6421 aatattggca attagccata ttagtcattg gttatatagc ataaatcaat attggctatt
     6481 ggccattgca tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa
     6541 tatgaccgcc atgttgacat tgattattga ctagttatta atagtaatca attacggggt
     6601 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     6661 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     6721 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     6781 acttggcagt acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg
     6841 gtaaatggcc cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc
     6901 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacacca
     6961 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     7021 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aataaccccg
     7081 ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
     7141 gtttagtgaa ccgtcagaat taaccatgga ctacaaagac catgacggtg attataaaga
     7201 tcatgacatc gattacaagg atgacgatga caagcttgcg gccgcgaatt caatggcggc
     7261 ggcggcgggc gcccctccgc cgggtccccc gcaaccgcct ccgccgccgc cgcccgagga
     7321 gtcgtccgac agcgagcccg aggcggagcc cggctcccca cagaagctca tccgcaaggt
     7381 gtccacgtcg ggtcagatcc gacagaagac catcatcaaa gaggggatgc tgaccaaaca
     7441 gaacaattca ttccagcgat caaaaaggag atactttaag cttcgagggc gaacgcttta
     7501 ctatgccaaa acggcaaagt caatcatatt tgatgaggtg gatctgacag atgccagcgt
     7561 agctgaatcc agtaccaaaa acgtcaacaa cagttttacg gtcataactc catgcaggaa
     7621 gctcatcttg tgtgctgata acagaaaaga aatggaagat tggattgcag cattaaagac
     7681 tgtgcagaac agggagcact ttgagcccac ccagtacagc atggaccact tctcagggat
     7741 gcacaattgg tacgcctgtt cccacgcgag gccgacctac tgcaatgtgt gccgtgaggc
     7801 tctgtctggg gtcacgtcgc acgggctgtc ctgcgaggtg tgcaaattta aggcccacaa
     7861 gcgctgtgct gtgcgtgcaa ccaataactg caagtggacc acactggcct cgatcgggaa
     7921 ggacatcatt gaagatgcag atgggattgc aatgccccac cagtggttgg aaggaaacct
     7981 acctgtgagc gccaagtgca ctgtgtgcga caagacctgt ggcagtgtgc tgcgcctgca
     8041 ggactggcgc tgcctctggt gcaaggccat ggttcacaca tcgtgtaaag aatccttgct
     8101 gaccaagtgc ccacttgg
//
