LOCUS       sequence-419957-        5865 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses directed activation strategy TMPRSS2 in mammalian cells, 
            such as HEK293F- or CHO-cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5865)
  AUTHORS   Verhulst E, De Bruyn M, Berckmans P, Sim Y, Augustyns K, Pintelon I,
            Berg M, Van Wielendaele P, Lambeir AM, Sterckx YG, Nelissen I, De 
            Meester I
  TITLE     Human Transmembrane Serine Protease 2 (TMPRSS2) on Human Seminal 
            Fluid Extracellular Vesicles Is Proteolytically Active.
  JOURNAL   J Extracell Vesicles. 2025 Mar;14(3):e70061. doi: 
            10.1002/jev2.70061.
  PUBMED    40091430
REFERENCE   2  (bases 1 to 5865)
  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: "J Extracell
            Vesicles."; date: "2025-03"; volume: "14(3):e70061. doi"; pages: " 
            10.1002/jev2.70061"
COMMENT     Sequence Label: pHLsec-dasTMPRSS2
FEATURES             Location/Qualifiers
     source          1..5865
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..662
                     /label=chicken beta-actin promoter
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1687..1706
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1761..1770
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2295..2309
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             3027..3071
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3084..3107
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(3219..3238)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3284..3339
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3338..3357)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3700..3716)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3700..3716)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3713..3735)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3724..3740
                     /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(3748..3754,3755..3772,3773..3778))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3793..3814
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3943..3960)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4114..4702)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4194..4213)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4873..5664,5665..5733))
                     /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     5496..5515
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5734..5838)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg ggggggcgcg cgccgggggg gggggggsgg ggsgaggggc
      541 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttgcggccgt
     1741 ctcaggccga attcaagctt gccaccatgg ggatccttcc cagccctggg atgcctgcgc
     1801 tgctctccct cgtgagcctt ctctccgtgc tgctgatggg ttgcgtagct gaaaccggtg
     1861 tgaagttcat gggcagcaag tgctccaact ctgggataga gtgcgactcc tcaggtacct
     1921 gcatcaaccc ctctaactgg tgtgatggcg tgtcacactg ccccggcggg gaggacgaga
     1981 atcggtgtgt tcgcctctac ggaccaaact tcatccttca ggtgtactca tctcagagga
     2041 agtcctggca ccctgtgtgc caagacgact ggaacgagaa ctacgggcgg gcggcctgca
     2101 gggacatggg ctataagaat aatttttact ctagccaagg aatagtggat gacagcggat
     2161 ccaccagctt tatgaaactg aacacaagtg ccggcaatgt cgatatctat aaaaaactgt
     2221 accacagtga tgcctgttct tcaaaagcag tggtttcttt acgctgtata gcctgcgggg
     2281 tcaacttgaa ctcagacgac gatgacaaga ttgtgggcgg cgagagcgcg ctcccggggg
     2341 cctggccctg gcaggtcagc ctgcacgtcc agaacgtcca cgtgtgcgga ggctccatca
     2401 tcacccccga gtggatcgtg acagccgccc actgcgtgga aaaacctctt aacaatccat
     2461 ggcattggac ggcatttgcg gggattttga gacaatcttt catgttctat ggagccggat
     2521 accaagtaga aaaagtgatt tctcatccaa attatgactc caagaccaag aacaatgaca
     2581 ttgcgctgat gaagctgcag aagcctctga ctttcaacga cctagtgaaa ccagtgtgtc
     2641 tgcccaaccc aggcatgatg ctgcagccag aacagctctg ctggatttcc gggtgggggg
     2701 ccaccgagga gaaagggaag acctcagaag tgctgaacgc tgccaaggtg cttctcattg
     2761 agacacagag atgcaacagc agatatgtct atgacaacct gatcacacca gccatgatct
     2821 gtgccggctt cctgcagggg aacgtcgatt cttgccaggg tgacagtgga gggcctctgg
     2881 tcacttcgaa gaacaatatc tggtggctga taggggatac aagctggggt tctggctgtg
     2941 ccaaagctta cagaccagga gtgtacggga atgtgatggt attcacggac tggatttatc
     3001 gacaaatgag ggcagacggc ggttcgggtc tgaacgacat ctttgaggcg caaaagattg
     3061 aatggcacga aggtcgcacc aaacaccacc atcaccatca ccaccactaa tagtgactcg
     3121 agactagtat cgcgataatt cactcctcag gtgcaggctg cctatcagaa ggtggtggct
     3181 ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat
     3241 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
     3301 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
     3361 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat
     3421 atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc
     3481 cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata
     3541 ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta
     3601 gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg
     3661 agatccctcg acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     3721 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     3781 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     3841 ccagtcggga aacctgtcgt gccagcggat cgatccgctg cattaatgaa tcggccaacg
     3901 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3961 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4021 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4081 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4141 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4201 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4261 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4321 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4381 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4441 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4501 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4561 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4621 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4681 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4741 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4801 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4861 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4921 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4981 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5041 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5101 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5161 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5221 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5281 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5341 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5401 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5461 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5521 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5581 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5641 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5701 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5761 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5821 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgg
//
