LOCUS       sequence-173602-        7151 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human DAGLb protein with intracellular V5 tag located on 
            the C-terminal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7151)
  AUTHORS   Singh PK, Markwick R, Lu L, Howell FV, Williams G, Doherty P
  TITLE     Assay and Inhibition of the Purified Catalytic Domain of 
            Diacylglycerol Lipase Beta.
  JOURNAL   Biochemistry. 2016 May 17;55(19):2713-21. doi: 
            10.1021/acs.biochem.6b00221. Epub 2016 May 4.
  PUBMED    27115711
REFERENCE   2  (bases 1 to 7151)
  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: 
            "10.1021/acs.biochem.6b00221"; journalName: "Biochemistry"; date: 
            "2016-05-17- 17"; volume: "55"; issue: "19"; pages: "2713-21"
COMMENT     Sequence Label: hDAGLb-V5
FEATURES             Location/Qualifiers
     source          1..7151
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    911..935
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2958..2982)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3008..3049
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3089..3108)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3133..3181
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3383..3811
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3470..3489)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3680..3701
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3820..3840)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3825..4154
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4005..4140
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4067..4086
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4202..4249
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4268..4666
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4824..4945
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4861..4880)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4915..4934
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4994..5010)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4994..5010)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5007..5029)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5018..5034
                     /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(5042..5048,5049..5066,5067..5072))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5087..5108
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5225..5242)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5396..5984)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5476..5495)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6155..6946,6947..7015))
                     /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     6778..6797
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7016..7120)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagctatca acaagtttgt acaaaaaagc aggcttcacc atgccgggga tggtactctt
      961 cggccggcgc tgggccatcg ccagcgacga cttggtcttc ccagggttct tcgagctggt
     1021 cgtgcgagtg ctgtggtgga ttggcattct gacgttgtat ctcatgcaca gaggaaagct
     1081 ggactgtgct ggtggagcct tgctcagcag ttacttgatc gtcctcatga ttctcctggc
     1141 agttgtcata tgtactgtgt cagccatcat gtgtgtcagc atgagaggaa cgatttgtaa
     1201 ccctggaccg cggaagtcta tgtctaagct gctttacatc cgcctggcgc tgttttttcc
     1261 agagatggtc tgggcctctc tgggggctgc ctgggtggca gatggtgttc agtgcgacag
     1321 gacagttgta aacggcatca tcgcaaccgt cgtggtcagt tggatcatca tcgctgccac
     1381 agtggtttcc attatcattg tctttgaccc tcttgggggg aaaatggctc catattcctc
     1441 tgccggcccc agccacctgg atagtcatga ttcaagccag ttacttaatg gcctcaagac
     1501 agcagctaca agcgtgtggg aaaccagaat caagctcttg tgctgttgca ttgggaaaga
     1561 cgaccatact cgggttgctt tttcgagtac ggcagagctt ttctcaacct acttttcaga
     1621 cacagatctg gtgcccagcg acattgcggc gggcctcgcc ctgcttcatc agcaacagga
     1681 caatatcagg aacaaccaag agcctgccca ggtggtctgc catgccccag ggagctccca
     1741 ggaagctgat ctggatgcag aattagaaaa ctgccatcat tacatgcagt ttgcagcagc
     1801 ggcctatggg tggcccctct acatctacag aaaccccctc acggggctgt gcaggattgg
     1861 tggtgactgc tgcagaagca gaaccacaga ctatgacttg gtcggaggcg atcagctcaa
     1921 ctgtcacttc ggctccatcc tgcacaccac agggctgcag tacagggact tcatccacgt
     1981 cagcttccat gacaaggttt acgagctgcc gtttttagtg gctctggatc acaggaaaga
     2041 gtctgttgtg gtcgctgtga gggggaccat gtctctgcag gatgtcctta cggacctgtc
     2101 agcggagagt gaggtgctgg acgtggagtg tgaggtgcag gaccgcctgg cacacaaggg
     2161 tatttctcaa gctgccagat acgtttacca acgactcatc aacgacggga ttttgagcca
     2221 agccttcagc attgctcctg agtaccggct ggtcatagtg ggccacagcc tcgggggagg
     2281 ggcggccgcc ctgctggcca ccatgctcag agccgcctac ccgcaggtca ggtgctacgc
     2341 cttctcccca ccccgggggc tgtggagcaa agctctgcag gaatattctc agagcttcat
     2401 cgtgtcactc gtcctgggga aggatgtgat tcccaggctc agtgtgacca acttggaaga
     2461 tctgaagaga agaatcttgc gagtggtcgc gcactgcaat aaacccaagt acaagatctt
     2521 gctgcacggt ttgtggtacg aactgtttgg aggaaacccc aacaacttgc ccacggagct
     2581 ggacgggggc gaccaggaag tcctgacaca gcctcttctg ggggagcaga gcctactgac
     2641 gcgctggtcc ccggcctaca gcttctccag cgactcccca ctggactctt ctcccaagta
     2701 cccccctctc taccctcccg gcaggatcat ccacctgcag gaggagggcg cctcggggcg
     2761 gtttggctgc tgctctgctg ctcactatag cgccaagtgg tcacacgaag cggaattcag
     2821 caaaatactc ataggtccga agatgctcac cgaccacatg ccagacatcc tgatgcgggc
     2881 cttggacagc gtggtctccg acagagcggc ctgcgtctcc tgtccagcac gaggggtctc
     2941 cagtgtggac gtggcccacc cagctttctt gtacaaagtg gttgatctag agggcccgcg
     3001 gttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc gtaccggtta
     3061 gtaatgagtt taaacggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     3121 cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt
     3181 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     3241 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     3301 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc
     3361 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3421 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3481 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3541 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3601 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3661 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3721 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3781 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3841 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3901 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3961 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4021 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4081 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4141 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg
     4201 tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa
     4261 ctaaaccatg gccaagcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc
     4321 tacaatcaac agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag
     4381 cgacggccgc atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga
     4441 actcgtggtg ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc
     4501 gatcggaaat gagaacaggg gcatcttgag cccctgcgga cggtgccgac aggtgcttct
     4561 cgatctgcat cctgggatca aagccatagt gaaggacagt gatggacagc cgacggcagt
     4621 tgggattcgt gaattgctgc cctctggtta tgtgtgggag ggctaagcac ttcgtggccg
     4681 aggagcagga ctgacacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt
     4741 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4801 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     4861 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4921 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     4981 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     5041 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     5101 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5161 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5221 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5281 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5341 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5401 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5461 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5521 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5581 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5641 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5701 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5761 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5821 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5881 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     5941 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6001 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6061 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6121 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6181 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6241 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6301 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6361 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6421 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6481 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6541 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6601 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6661 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6721 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6781 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6841 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6901 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     6961 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7021 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7081 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7141 cacctgacgt c
//
