LOCUS       sequence-83086-e        8802 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses full-length Endothelin converting enzyme 1 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 8802)
  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 8802)
  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: ECE1-His-bio
FEATURES             Location/Qualifiers
     source          1..8802
                     /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    2902..2957
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2956..2975)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5193..5211)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5279..5383
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5384..5452,5453..6244)
                     /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(5602..5621)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6415..7003
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6904..6923
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7157..7174
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7291..7312
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7327..7332,7333..7350,7351..7357)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7365..7381
                     /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     7370..7392
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7389..7405
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7389..7405
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7676..8055
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8056..8259
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8210..8230
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8256..8280
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     8761..8780
                     /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 ccgcccggaa tccagtacca gaccagatcc cccagcgtgt gcctgagcga ggcctgtgtg
      841 tccgtgacca gcagcatcct gagcagcatg gaccccaccg tggacccctg ccacgacttc
      901 ttcagctacg cctgcggcgg ctggatcaag gccaaccctg tgcccgacgg ccacagcaga
      961 tggggcacct tcagcaacct gtgggagcac aaccaggcca tcatcaaaca tctgctggaa
     1021 aacagcaccg ccagcgtgtc cgaggccgag agaaaggccc aggtgtacta ccgggcctgc
     1081 atgaacgaga cacggatcga ggaactgcgg gccaagcccc tgatggaact gatcgagcgg
     1141 ctgggcggct ggaacatcac aggcccctgg gccaaggaca acttccagga caccctgcag
     1201 gtcgtgaccg cccactaccg gaccagccca ttcttcagcg tgtacgtgtc cgccgacagc
     1261 aagaacagca acagcaacgt gatccaggtg gaccagagcg gcctgggcct gcccagcaga
     1321 gactactacc tgaacaagac cgagaacgag aaggtgctga ccggctacct gaactacatg
     1381 gtgcagctgg gcaagctgct gggcggaggc gacgaggaag ccatcagacc ccagatgcag
     1441 cagatcctgg acttcgagac agccctggcc aacatcacca tcccccagga aaagcggcgg
     1501 gacgaggaac tgatctacca caaagtgaca gccgccgagc tgcagaccct ggcccctgct
     1561 atcaactggc tgcccttcct gaacaccatc ttctaccccg tggaaatcaa cgagagcgag
     1621 cccatcgtgg tgtacgacaa agagtacctg gaacagatca gcaccctgat caacaccacc
     1681 gaccggtgcc tgctgaacaa ctacatgatc tggaacctcg tgcgcaagac ctccagcttt
     1741 ctggaccagc ggtttcagga cgccgacgag aagttcatgg aagtgatgta cggcaccaag
     1801 aaaacctgcc tgccccggtg gaagttctgc gtgtccgaca ccgagaacaa cctgggcttc
     1861 gccctgggcc ccatgttcgt gaaggccacc ttcgccgagg acagcaagtc tatcgccacc
     1921 gagatcatcc tcgagatcaa gaaggccttc gaggaaagcc tgagcaccct gaagtggatg
     1981 gacgaggaaa ccagaaagag cgccaaagag aaggccgacg ccatctacaa catgatcggc
     2041 taccccaact tcatcatgga ccctaaagaa ctggacaagg tgttcaacga ctacaccgcc
     2101 gtgcccgacc tgtacttcga gaacgccatg cggttcttca acttctcctg gcgcgtgacc
     2161 gccgaccagc tgaggaaggc ccccaacagg gaccagtggt ccatgacccc ccccatggtc
     2221 aacgcctact acagccccac caagaacgag atcgtgttcc ctgccggcat cctgcaggcc
     2281 ccattctaca ccagaagcag ccccaaggcc ctgaacttcg gcggcatcgg cgtggtcgtg
     2341 ggccacgaac tgacccacgc cttcgacgac cagggcagag agtacgacaa ggacggcaac
     2401 ctgcggccct ggtggaagaa cagcagcgtg gaagccttca agcggcagac cgagtgcatg
     2461 gtggaacagt acagcaacta cagcgtgaac ggcgagcccg tgaacggcag acacaccctg
     2521 ggcgagaata tcgccgacaa tggcggactg aaggccgcct acagagccta ccagaactgg
     2581 gtcaagaaga acggcgccga gcacagcctg cccaccctgg gcctgaccaa caaccagctg
     2641 ttcttcctgg gattcgcaca ggtctggtgc agcgtgcgga cccctgagtc tagccacgag
     2701 ggcctgatca ccgaccccca cagccccagc cggttcagag tgatcggcag cctgagcaac
     2761 tccaaagagt tcagcgagca cttccggtgc ccccctggca gccccatgaa ccctccccac
     2821 aagtgcgaag tgtggtgagg cgcgcctgaa ttcttctaga cacgtgatta attaaggatc
     2881 ccccgacctc gacctctggc taataaagga aatttatttt cattgcaata gtgtgttgga
     2941 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcatttg gtcgagatcc
     3001 ctcggagatc tctagctaga ggatcgatcc ccgccccgga cgaactaaac ctgactacga
     3061 catctctgcc ccttcttcgc ggggcagtgc atgtaatccc ttcagttggt tggtacaact
     3121 tgccaactgg gccctgttcc acatgtgaca cgggggggga ccaaacacaa aggggttctc
     3181 tgactgtagt tgacatcctt ataaatggat gtgcacattt gccaacactg agtggctttc
     3241 atcctggagc agactttgca gtctgtggac tgcaacacaa cattgccttt atgtgtaact
     3301 cttggctgaa gctcttacac caatgctggg ggacatgtac ctcccagggg cccaggaaga
     3361 ctacgggagg ctacaccaac gtcaatcaga ggggcctgtg tagctaccga taagcggacc
     3421 ctcaagaggg cattagcaat agtgtttata aggccccctt gttaacccta aacgggtagc
     3481 atatgcttcc cgggtagtag tatatactat ccagactaac cctaattcaa tagcatatgt
     3541 tacccaacgg gaagcatatg ctatcgaatt agggttagta aaagggtcct aaggaacagc
     3601 gatatctccc accccatgag ctgtcacggt tttatttaca tggggtcagg attccacgag
     3661 ggtagtgaac cattttagtc acaagggcag tggctgaaga tcaaggagcg ggcagtgaac
     3721 tctcctgaat cttcgcctgc ttcttcattc tccttcgttt agctaataga ataactgctg
     3781 agttgtgaac agtaaggtgt atgtgaggtg ctcgaaaaca aggtttcagg tgacgccccc
     3841 agaataaaat ttggacgggg ggttcagtgg tggcattgtg ctatgacacc aatataaccc
     3901 tcacaaaccc cttgggcaat aaatactagt gtaggaatga aacattctga atatctttaa
     3961 caatagaaat ccatggggtg gggacaagcc gtaaagactg gatgtccatc tcacacgaat
     4021 ttatggctat gggcaacaca taatcctagt gcaatatgat actggggtta ttaagatgtg
     4081 tcccaggcag ggaccaagac aggtgaacca tgttgttaca ctctatttgt aacaagggga
     4141 aagagagtgg acgccgacag cagcggactc cactggttgt ctctaacacc cccgaaaatt
     4201 aaacggggct ccacgccaat ggggcccata aacaaagaca agtggccact cttttttttg
     4261 aaattgtgga gtgggggcac gcgtcagccc ccacacgccg ccctgcggtt ttggactgta
     4321 aaataagggt gtaataactt ggctgattgt aaccccgcta accactgcgg tcaaaccact
     4381 tgcccacaaa accactaatg gcaccccggg gaatacctgc ataagtaggt gggcgggcca
     4441 agataggggc gcgattgctg cgatctggag gacaaattac acacacttgc gcctgagcgc
     4501 caagcacagg gttgttggtc ctcatattca cgaggtcgct gagagcacgg tgggctaatg
     4561 ttgccatggg tagcatatac tacccaaata tctggatagc atatgctatc ctaatctata
     4621 tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata
     4681 tctgggtagt atatgctatc ctaatttata tctgggtagc ataggctatc ctaatctata
     4741 tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc ctaatctgta
     4801 tccgggtagc atatgctatc ctaatagaga ttagggtagt atatgctatc ctaatttata
     4861 tctgggtagc atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg
     4921 ggtagcatat gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg
     4981 ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg
     5041 ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg
     5101 ggtagtatat gctatcctaa tctgtatccg ggtagcatat gctatcctca tgataagctg
     5161 tcaaacatga gaattttctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     5221 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     5281 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     5341 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     5401 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     5461 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     5521 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     5581 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     5641 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     5701 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5761 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5821 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5881 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     5941 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     6001 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     6061 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     6121 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     6181 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     6241 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     6301 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6361 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6421 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6481 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6541 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     6601 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6661 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6721 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6781 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6841 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6901 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6961 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     7021 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     7081 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     7141 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     7201 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     7261 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     7321 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     7381 atttcacaca ggaaacagct atgaccatga ttacgccaag ctctagctag aggtcgacca
     7441 attctcatgt ttgacagctt atcatcgcag atccgggcaa cgttgttgcc attgctgcag
     7501 gcgcagaact ggtaggtatg gaagatctat acattgaatc aatattggca attagccata
     7561 ttagtcattg gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat
     7621 ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc atgttgacat
     7681 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     7741 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     7801 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     7861 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     7921 tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     7981 tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta cgtattagtc
     8041 atcgctatta ccatggtgat gcggttttgg cagtacacca atgggcgtgg atagcggttt
     8101 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     8161 caaaatcaac gggactttcc aaaatgtcgt aataaccccg ccccgttgac gcaaatgggc
     8221 ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc
     8281 ctcactctct tccgcatcgc tgtctgcgag ggccagctgt tgggctcgcg gttgaggaca
     8341 aactcttcgc ggtctttcca gtactcttgg atcggaaacc cgtcggcctc cgaacggtac
     8401 tccgccaccg agggacctga gcgagtccgc atcgaccgga tcggaaaacc tctcgagaaa
     8461 ggcgtctaac cagtcacagt cgcaaggtag gctgagcacc gtggcgggcg gcagcgggtg
     8521 gcggtcgggg ttgtttctgg cggaggtgct gctgatgatg taattaaagt aggcggtctt
     8581 gagacggcgg atggtcgagg tgaggtgtgg caggcttgag atccagctgt tggggtgagt
     8641 actccctctc aaaagcgggc attacttctg cgctaagatt gtcagtttcc aaaaacgagg
     8701 aggatttgat attcacctgg cccgatctgg ccatacactt gagtgacaat gacatccact
     8761 ttgcctttct ctccacaggt gtccactccc aggtccaagt tt
//
