LOCUS       sequence-81058-e        7398 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses full-length Tumor necrosis factor ligand superfamily 
            member 5 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 7398)
  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 7398)
  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: CD154-His-bio
FEATURES             Location/Qualifiers
     source          1..7398
                     /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    1498..1553
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1552..1571)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3789..3807)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3875..3979
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3980..4048,4049..4840)
                     /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(4198..4217)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5011..5599
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5500..5519
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5753..5770
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5887..5908
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5923..5928,5929..5946,5947..5953)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5961..5977
                     /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     5966..5988
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5985..6001
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5985..6001
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        6272..6651
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6652..6855
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6806..6826
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6852..6876
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     7357..7376
                     /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 ccgcctcaca gacggctgga caagatcgag gacgagcgga acctgcacga ggacttcgtg
      841 ttcatgaaga ccatccagcg gtgcaacacc ggcgagcgga gcctgagcct gctgaactgc
      901 gaggaaatca agagccagtt cgagggcttc gtgaaggaca tcatgctgaa caaagaggaa
      961 actaagaaag aaaacagctt cgagatgcag aagggcgacc agaaccccca gatcgccgcc
     1021 cacgtgatca gcgaggccag cagcaagacc accagcgtgc tgcagtgggc cgagaagggc
     1081 tactacacca tgagcaacaa cctggtcacc ctggaaaacg gcaagcagct gaccgtgaag
     1141 cggcagggcc tgtactacat ctacgcccag gtcacattct gcagcaacag agaggccagc
     1201 tcccaggccc cctttatcgc cagcctgtgc ctgaagtccc ccggcagatt cgagcgcatc
     1261 ctgctgagag ccgccaacac ccacagcagc gccaagcctt gcggccagca gagcatccac
     1321 ctcggaggcg tgttcgagct gcagcctggc gcctctgtgt tcgtgaacgt gaccgacccc
     1381 agccaggtgt cccacggcac cggattcacc agcttcggcc tgctgaagct ctgaggcgcg
     1441 cctgaattct tctagacacg tgattaatta aggatccccc gacctcgacc tctggctaat
     1501 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     1561 ggacatatgg gagggcaaat catttggtcg agatccctcg gagatctcta gctagaggat
     1621 cgatccccgc cccggacgaa ctaaacctga ctacgacatc tctgcccctt cttcgcgggg
     1681 cagtgcatgt aatcccttca gttggttggt acaacttgcc aactgggccc tgttccacat
     1741 gtgacacggg gggggaccaa acacaaaggg gttctctgac tgtagttgac atccttataa
     1801 atggatgtgc acatttgcca acactgagtg gctttcatcc tggagcagac tttgcagtct
     1861 gtggactgca acacaacatt gcctttatgt gtaactcttg gctgaagctc ttacaccaat
     1921 gctgggggac atgtacctcc caggggccca ggaagactac gggaggctac accaacgtca
     1981 atcagagggg cctgtgtagc taccgataag cggaccctca agagggcatt agcaatagtg
     2041 tttataaggc ccccttgtta accctaaacg ggtagcatat gcttcccggg tagtagtata
     2101 tactatccag actaacccta attcaatagc atatgttacc caacgggaag catatgctat
     2161 cgaattaggg ttagtaaaag ggtcctaagg aacagcgata tctcccaccc catgagctgt
     2221 cacggtttta tttacatggg gtcaggattc cacgagggta gtgaaccatt ttagtcacaa
     2281 gggcagtggc tgaagatcaa ggagcgggca gtgaactctc ctgaatcttc gcctgcttct
     2341 tcattctcct tcgtttagct aatagaataa ctgctgagtt gtgaacagta aggtgtatgt
     2401 gaggtgctcg aaaacaaggt ttcaggtgac gcccccagaa taaaatttgg acggggggtt
     2461 cagtggtggc attgtgctat gacaccaata taaccctcac aaaccccttg ggcaataaat
     2521 actagtgtag gaatgaaaca ttctgaatat ctttaacaat agaaatccat ggggtgggga
     2581 caagccgtaa agactggatg tccatctcac acgaatttat ggctatgggc aacacataat
     2641 cctagtgcaa tatgatactg gggttattaa gatgtgtccc aggcagggac caagacaggt
     2701 gaaccatgtt gttacactct atttgtaaca aggggaaaga gagtggacgc cgacagcagc
     2761 ggactccact ggttgtctct aacacccccg aaaattaaac ggggctccac gccaatgggg
     2821 cccataaaca aagacaagtg gccactcttt tttttgaaat tgtggagtgg gggcacgcgt
     2881 cagcccccac acgccgccct gcggttttgg actgtaaaat aagggtgtaa taacttggct
     2941 gattgtaacc ccgctaacca ctgcggtcaa accacttgcc cacaaaacca ctaatggcac
     3001 cccggggaat acctgcataa gtaggtgggc gggccaagat aggggcgcga ttgctgcgat
     3061 ctggaggaca aattacacac acttgcgcct gagcgccaag cacagggttg ttggtcctca
     3121 tattcacgag gtcgctgaga gcacggtggg ctaatgttgc catgggtagc atatactacc
     3181 caaatatctg gatagcatat gctatcctaa tctatatctg ggtagcatag gctatcctaa
     3241 tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa
     3301 tttatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa
     3361 tctatatctg ggtagtatat gctatcctaa tctgtatccg ggtagcatat gctatcctaa
     3421 tagagattag ggtagtatat gctatcctaa tttatatctg ggtagcatat actacccaaa
     3481 tatctggata gcatatgcta tcctaatcta tatctgggta gcatatgcta tcctaatcta
     3541 tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta
     3601 tatctgggta gtatatgcta tcctaattta tatctgggta gcataggcta tcctaatcta
     3661 tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaatctg
     3721 tatccgggta gcatatgcta tcctcatgat aagctgtcaa acatgagaat tttcttgaag
     3781 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     3841 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3901 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3961 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4021 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4081 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4141 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4201 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
     4261 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4321 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4381 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4441 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4501 cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg
     4561 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     4621 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4681 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4741 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     4801 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     4861 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     4921 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     4981 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5041 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5101 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5161 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5221 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5281 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5341 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5401 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5461 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5521 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5581 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5641 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     5701 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     5761 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     5821 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     5881 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     5941 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     6001 ccatgattac gccaagctct agctagaggt cgaccaattc tcatgtttga cagcttatca
     6061 tcgcagatcc gggcaacgtt gttgccattg ctgcaggcgc agaactggta ggtatggaag
     6121 atctatacat tgaatcaata ttggcaatta gccatattag tcattggtta tatagcataa
     6181 atcaatattg gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat
     6241 attggctcat gtccaatatg accgccatgt tgacattgat tattgactag ttattaatag
     6301 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     6361 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     6421 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     6481 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct
     6541 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg
     6601 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     6661 ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     6721 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     6781 tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     6841 tatataagca gagctcgttt agtgaaccgt cagatcctca ctctcttccg catcgctgtc
     6901 tgcgagggcc agctgttggg ctcgcggttg aggacaaact cttcgcggtc tttccagtac
     6961 tcttggatcg gaaacccgtc ggcctccgaa cggtactccg ccaccgaggg acctgagcga
     7021 gtccgcatcg accggatcgg aaaacctctc gagaaaggcg tctaaccagt cacagtcgca
     7081 aggtaggctg agcaccgtgg cgggcggcag cgggtggcgg tcggggttgt ttctggcgga
     7141 ggtgctgctg atgatgtaat taaagtaggc ggtcttgaga cggcggatgg tcgaggtgag
     7201 gtgtggcagg cttgagatcc agctgttggg gtgagtactc cctctcaaaa gcgggcatta
     7261 cttctgcgct aagattgtca gtttccaaaa acgaggagga tttgatattc acctggcccg
     7321 atctggccat acacttgagt gacaatgaca tccactttgc ctttctctcc acaggtgtcc
     7381 actcccaggt ccaagttt
//
