LOCUS       sequence-305775-        6627 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of human Par2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6627)
  AUTHORS   Bohm SK, Kong W, Bromme D, Smeekens SP, Anderson DC, Connolly A, 
            Kahn M, Nelken NA, Coughlin SR, Payan DG, Bunnett NW
  TITLE     Molecular cloning, expression and potential functions of the human 
            proteinase-activated receptor-2.
  JOURNAL   Biochem J. 1996 Mar 15;314 ( Pt 3)(Pt 3):1009-16. doi: 
            10.1042/bj3141009.
  PUBMED    8615752
REFERENCE   2  (bases 1 to 6627)
  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: "Biochem 
            J."; date: "1996-03-15"; volume: "314 ( Pt 3)(Pt 3):1009-16. doi"; 
            pages: " 10.1042/bj3141009"
COMMENT     Sequence Label: pcDNA3-hPar2
FEATURES             Location/Qualifiers
     source          1..6627
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      661..670
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             670..693
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1882..1908
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(1946..1964)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1947..1964)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1984..2001)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1990..2214
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2260..2688
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2347..2366)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2557..2578
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2697..2717)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2702..3031
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2882..3017
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2944..2963
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3098..3892
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3152..3171)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3762..3781
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4066..4187
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4103..4122)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4157..4176
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4236..4252)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4236..4252)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4249..4271)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4260..4276
                     /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(4284..4290,4291..4308,4309..4314))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4329..4350
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4467..4484)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4638..5226)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4718..4737)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5397..6188,6189..6257))
                     /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     6020..6039
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6258..6362)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6437..6456)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 gccaccatgg actacaagga cgacgacgac aagatgcgga gccccagcgc ggcgtggctg
      721 ctgggggccg ccatcctgct agcagcctct ctctcctgca gtggcaccat ccaaggaacc
      781 aatagatcct ctaaaggaag aagccttatt ggtaaggttg atggcacatc ccacgtcact
      841 ggaaaaggag ttacagttga aacagtcttt tctgtggatg agttttctgc atctgtcctc
      901 actggaaaac tgaccactgt cttccttcca attgtctaca caattgtgtt tgtggtgggt
      961 ttgccaagta acggcatggc cctgtgggtc tttcttttcc gaactaagaa gaagcaccct
     1021 gctgtgattt acatggccaa tctggccttg gctgacctcc tctctgtcat ctggttcccc
     1081 ttgaagattg cctatcacat acatggcaac aactggattt atggggaagc tctttgtaat
     1141 gtgcttattg gctttttcta tggcaacatg tactgttcca ttctcttcat gacctgcctc
     1201 agtgtgcaga ggtattgggt catcgtgaac cccatggggc actccaggaa gaaggcaaac
     1261 attgccattg gcatctccct ggcaatatgg ctgctgattc tgctggtcac catccctttg
     1321 tatgtcgtga agcagaccat cttcattcct gccctgaaca tcacgacctg tcatgatgtt
     1381 ttgcctgagc agctcttggt gggagacatg ttcaattact tcctctctct ggccattggg
     1441 gtctttctgt tcccagcctt cctcacagcc tctgcctatg tgctgatgat cagaatgctg
     1501 cgatcttctg ccatggatga aaactcagag aagaaaagga agagggccat caaactcatt
     1561 gtcactgtcc tggccatgta cctgatctgc ttcactccta gtaaccttct gcttgtggtg
     1621 cattattttc tgattaagag ccagggccag agccatgtct atgccctgta cattgtagcc
     1681 ctctgcctct ctacccttaa cagctgcatc gacccctttg tctattactt tgtttcacat
     1741 gatttcaggg atcatgcaaa gaacgctctc ctttgccgaa gtgtccgcac tgtaaagcag
     1801 atgcaagtat ccctcacctc aaagaaacac tccaggaaat ccagctctta ctcttcaagt
     1861 tcaaccactg ttaagacctc ctatccctat gatgttcccg attatgccta ggcggccgct
     1921 cgagcatgca tctagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat
     1981 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     2041 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     2101 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     2161 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     2221 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     2281 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     2341 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2401 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     2461 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2521 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     2581 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     2641 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     2701 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2761 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     2821 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     2881 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     2941 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     3001 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     3061 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     3121 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3181 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     3241 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     3301 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     3361 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     3421 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     3481 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     3541 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     3601 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     3661 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     3721 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     3781 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     3841 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     3901 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     3961 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4021 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     4081 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4141 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     4201 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4261 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4321 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4381 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4441 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4501 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4561 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4621 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4681 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4741 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4801 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4861 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4921 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4981 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5041 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5101 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5161 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5221 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5281 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5341 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5401 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5461 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5521 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5581 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5641 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5701 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5761 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5821 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5881 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5941 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6001 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6061 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6121 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6181 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6241 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6301 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6361 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     6421 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     6481 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     6541 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc
     6601 gcgatgtacg ggccagatat acgcgtt
//
