LOCUS       sequence-83089-e        8184 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses full-length Leucine rich repeat containing 8 family, 
            member B ectodomain in mammalian cells. N-terminal His tag, 
            biotinylation sequence and rat Cd4d3+4 tag. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8184)
  AUTHORS   Sun Y, Vandenbriele C, Kauskot A, Verhamme P, Hoylaerts MF, Wright 
            GJ
  TITLE     A human platelet receptor protein microarray identifies 
            FcepsilonR1alpha as an activating PEAR1 ligand.
  JOURNAL   Mol Cell Proteomics. 2015 Feb 23. pii: mcp.M114.046946.
  PUBMED    25713122
REFERENCE   2  (bases 1 to 8184)
  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"; journalName: "Mol Cell 
            Proteomics. 2015 Feb 23. pii: mcp.M114.046946."
COMMENT     Sequence Label: LRRC8b-His-bio
FEATURES             Location/Qualifiers
     source          1..8184
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..10
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             70..87
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2284..2339
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2338..2357)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4575..4593)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4661..4765
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4766..4834,4835..5626)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4984..5003)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5797..6385
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6286..6305
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6539..6556
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6673..6694
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6709..6714,6715..6732,6733..6739)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6747..6763
                     /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     6752..6774
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6771..6787
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6771..6787
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7058..7437
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7438..7641
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7592..7612
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7638..7662
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     8143..8162
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 gccaccatgg agtttcagac ccaggtactc atgtccctgc tgctctgcat gtctggtctg
       61 aattccggcc accaccatca ccaccatagc cccgacagga acctgcctcc tctggcccct
      121 ctgggcccca acagcggcag cccagacctg catcatattc tggatgcaca gaaaatggtg
      181 tggaatcatc gtgacagaaa cctgcctcca ctagcacctc tgtcgacctc catcacggcc
      241 tataagagtg agggggagtc agcggagttc tccttcccac tcaaccttgg agaggaaagc
      301 ctgcagggag agttgagatg gaaggcagag aaggctcctt cttcccagtc ctggatcacc
      361 ttctccctaa agaaccaaaa ggtgtctgtg cagaagtcta ctagcaaccc caagttccag
      421 ctgtccgaaa cgctcccact cacccttcag ataccccagg tctcccttca gtttgctggt
      481 tctggcaacc tgaccctgac tctggacaga gggatactgt atcaggaagt gaacctggtg
      541 gtgatgaaag tgactcagcc cgacagcaac actttgacct gtgaggtgat gggacccacc
      601 tcacccaaga tgagactgat cttgaagcag gagaatcagg aggccagggt ctccaggcag
      661 gagaaagtga ttcaagtgca ggcccctgaa gcaggggtgt ggcaatgtct actgagtgaa
      721 ggtgaagagg tcaagatgga ctccaagatc caggttttat ccaaagggtt gaattcgcgg
      781 ccgcctggcc tgacaagcag ctacagcctg tggtggatgc tgagatcttc cctgaaacag
      841 tacagcttcg aggccctgag agagaagtcc aactacagcg acatccccga cgtgaagaac
      901 gacttcgcct tcatcctgca cctggccgac cagtacgacc ccctgtacag caagcggttc
      961 agcatcttcc tgagcgaggt gtccgagaac aagctgaagc agatcaacct gaacaacgag
     1021 tggaccgtcg agaagctgaa gtccaagctg gtcaagaacg cccaggacaa gatcgagctg
     1081 cacctgttca tgctgaacgg cctgcccgac aacgtgttcg agctgaccga gatggaagtg
     1141 ctgagcctgg aactgatccc tgaagtgaag ctgcccagcg ccgtgtccca gctcgtgaac
     1201 ctgaaagaac tgcgggtgta ccacagctcc ctggtggtgg accaccccgc cctggccttt
     1261 ctggaagaga acctgaagat cctgcggctg aagttcaccg agatgggcaa gatccccaga
     1321 tgggtgttcc acctgaagaa tctgaaagag ctgtacctga gcggctgcgt gctgcccgag
     1381 cagctgagca ccatgcagct cgaaggcttt caggacctga aaaacctgcg gaccctgtac
     1441 ctgaagtcca gcctgagccg gatcccccag gtggtcaccg acctgctgcc cagcctgcag
     1501 aagctgtccc tggacaacga gggcagcaag ctggtggtgc tgaacaacct gaagaagatg
     1561 gtcaacctca agagcctcga gctgattagc tgcgacctgg aaagaatccc ccacagcatc
     1621 ttcagcctca acaatctcca cgagctggat ctgcgcgaga acaatctcaa gaccgtggaa
     1681 gagatcatca gcttccagca tctgcagaac ctgtcctgcc tgaagctgtg gcacaacaat
     1741 atcgcctaca tccccgccca gatcggcgcc ctgagcaacc tggaacagct ctctctggat
     1801 cacaacaaca tcgagaacct gcccctgcag ctgttcctgt gcaccaagct gcactacctg
     1861 gacctgagct acaaccacct gaccttcatc cccgaggaaa tccagtacct gtccaacctg
     1921 cagtacttcg ccgtgaccaa caacaatatt gagatgctgc ccgacggcct gttccagtgc
     1981 aagaaactgc agtgcctgct gctgggcaag aacagcctga tgaacctgag cccccacgtc
     2041 ggcgagctga gcaatctgac ccacctggaa ctcatcggca actacctgga aaccctgccc
     2101 cccgagctgg aaggatgcca gagcctgaag cggaactgcc tgatcgtgga agaaaacctg
     2161 ctgaacaccc tgcctctgcc cgtgaccgaa cggctccaga catgcctgga caagtgctga
     2221 ggcgcgcctg aattcttcta gacacgtgat taattaagga tcccccgacc tcgacctctg
     2281 gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac
     2341 tcggaaggac atatgggagg gcaaatcatt tggtcgagat ccctcggaga tctctagcta
     2401 gaggatcgat ccccgccccg gacgaactaa acctgactac gacatctctg ccccttcttc
     2461 gcggggcagt gcatgtaatc ccttcagttg gttggtacaa cttgccaact gggccctgtt
     2521 ccacatgtga cacggggggg gaccaaacac aaaggggttc tctgactgta gttgacatcc
     2581 ttataaatgg atgtgcacat ttgccaacac tgagtggctt tcatcctgga gcagactttg
     2641 cagtctgtgg actgcaacac aacattgcct ttatgtgtaa ctcttggctg aagctcttac
     2701 accaatgctg ggggacatgt acctcccagg ggcccaggaa gactacggga ggctacacca
     2761 acgtcaatca gaggggcctg tgtagctacc gataagcgga ccctcaagag ggcattagca
     2821 atagtgttta taaggccccc ttgttaaccc taaacgggta gcatatgctt cccgggtagt
     2881 agtatatact atccagacta accctaattc aatagcatat gttacccaac gggaagcata
     2941 tgctatcgaa ttagggttag taaaagggtc ctaaggaaca gcgatatctc ccaccccatg
     3001 agctgtcacg gttttattta catggggtca ggattccacg agggtagtga accattttag
     3061 tcacaagggc agtggctgaa gatcaaggag cgggcagtga actctcctga atcttcgcct
     3121 gcttcttcat tctccttcgt ttagctaata gaataactgc tgagttgtga acagtaaggt
     3181 gtatgtgagg tgctcgaaaa caaggtttca ggtgacgccc ccagaataaa atttggacgg
     3241 ggggttcagt ggtggcattg tgctatgaca ccaatataac cctcacaaac cccttgggca
     3301 ataaatacta gtgtaggaat gaaacattct gaatatcttt aacaatagaa atccatgggg
     3361 tggggacaag ccgtaaagac tggatgtcca tctcacacga atttatggct atgggcaaca
     3421 cataatccta gtgcaatatg atactggggt tattaagatg tgtcccaggc agggaccaag
     3481 acaggtgaac catgttgtta cactctattt gtaacaaggg gaaagagagt ggacgccgac
     3541 agcagcggac tccactggtt gtctctaaca cccccgaaaa ttaaacgggg ctccacgcca
     3601 atggggccca taaacaaaga caagtggcca ctcttttttt tgaaattgtg gagtgggggc
     3661 acgcgtcagc ccccacacgc cgccctgcgg ttttggactg taaaataagg gtgtaataac
     3721 ttggctgatt gtaaccccgc taaccactgc ggtcaaacca cttgcccaca aaaccactaa
     3781 tggcaccccg gggaatacct gcataagtag gtgggcgggc caagataggg gcgcgattgc
     3841 tgcgatctgg aggacaaatt acacacactt gcgcctgagc gccaagcaca gggttgttgg
     3901 tcctcatatt cacgaggtcg ctgagagcac ggtgggctaa tgttgccatg ggtagcatat
     3961 actacccaaa tatctggata gcatatgcta tcctaatcta tatctgggta gcataggcta
     4021 tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta
     4081 tcctaattta tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta
     4141 tcctaatcta tatctgggta gtatatgcta tcctaatctg tatccgggta gcatatgcta
     4201 tcctaataga gattagggta gtatatgcta tcctaattta tatctgggta gcatatacta
     4261 cccaaatatc tggatagcat atgctatcct aatctatatc tgggtagcat atgctatcct
     4321 aatctatatc tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct
     4381 aatctatatc tgggtagtat atgctatcct aatttatatc tgggtagcat aggctatcct
     4441 aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct
     4501 aatctgtatc cgggtagcat atgctatcct catgataagc tgtcaaacat gagaattttc
     4561 ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     4621 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4681 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4741 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4801 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4861 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4921 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4981 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
     5041 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5101 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5161 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5221 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5281 aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt
     5341 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5401 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5461 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5521 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5581 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     5641 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5701 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5761 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5821 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5881 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5941 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6001 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6061 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6121 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6181 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6241 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6301 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6361 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6421 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     6481 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     6541 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     6601 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     6661 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     6721 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     6781 ctatgaccat gattacgcca agctctagct agaggtcgac caattctcat gtttgacagc
     6841 ttatcatcgc agatccgggc aacgttgttg ccattgctgc aggcgcagaa ctggtaggta
     6901 tggaagatct atacattgaa tcaatattgg caattagcca tattagtcat tggttatata
     6961 gcataaatca atattggcta ttggccattg catacgttgt atctatatca taatatgtac
     7021 atttatattg gctcatgtcc aatatgaccg ccatgttgac attgattatt gactagttat
     7081 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     7141 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     7201 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     7261 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtccg
     7321 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     7381 ttacgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
     7441 atgcggtttt ggcagtacac caatgggcgt ggatagcggt ttgactcacg gggatttcca
     7501 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
     7561 ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg
     7621 gaggtctata taagcagagc tcgtttagtg aaccgtcaga tcctcactct cttccgcatc
     7681 gctgtctgcg agggccagct gttgggctcg cggttgagga caaactcttc gcggtctttc
     7741 cagtactctt ggatcggaaa cccgtcggcc tccgaacggt actccgccac cgagggacct
     7801 gagcgagtcc gcatcgaccg gatcggaaaa cctctcgaga aaggcgtcta accagtcaca
     7861 gtcgcaaggt aggctgagca ccgtggcggg cggcagcggg tggcggtcgg ggttgtttct
     7921 ggcggaggtg ctgctgatga tgtaattaaa gtaggcggtc ttgagacggc ggatggtcga
     7981 ggtgaggtgt ggcaggcttg agatccagct gttggggtga gtactccctc tcaaaagcgg
     8041 gcattacttc tgcgctaaga ttgtcagttt ccaaaaacga ggaggatttg atattcacct
     8101 ggcccgatct ggccatacac ttgagtgaca atgacatcca ctttgccttt ctctccacag
     8161 gtgtccactc ccaggtccaa gttt
//
