LOCUS       sequence-367374-        8949 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector - TMPRSS2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8949)
  AUTHORS   Kastenhuber ER, Mercadante M, Nilsson-Payant B, Johnson JL, Jaimes 
            JA, Muecksch F, Weisblum Y, Bram Y, Chandar V, Whittaker GR, 
            tenOever BR, Schwartz RE, Cantley L
  TITLE     Coagulation factors directly cleave SARS-CoV-2 spike and enhance 
            viral entry.
  JOURNAL   Elife. 2022 Mar 23;11. pii: 77444. doi: 10.7554/eLife.77444.
  PUBMED    35294338
REFERENCE   2  (bases 1 to 8949)
  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: "Elife."; 
            date: "2022-03-23"; volume: "11. pii: 77444. doi"; pages: " 
            10.7554/eLife.77444"
COMMENT     Sequence Label: pEGPN-TMPRSS2
FEATURES             Location/Qualifiers
     source          1..8949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1684..1917
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2102..2146
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2444..2561
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2657..2868
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2907..2916
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        4445..4944
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4480..4503)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4877..4896
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4981..5775
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4981..5775
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5035..5054)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5645..5664
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    5838..6426
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5891..5911)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6309..6320)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6429..6445)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6430..6446)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6955..7135
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7197..7785)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7277..7296)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7956..8747,8748..8816))
                     /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     8579..8598
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8817..8921)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga
     1561 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     1621 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     1681 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     1741 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     1801 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     1861 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     1921 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     1981 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     2041 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     2101 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     2161 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     2221 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     2281 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
     2341 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact
     2401 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg
     2461 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     2521 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     2581 gcagagatcc agtttggtta gtaccgggcc cggggatcag tgtgagggag tgtaaagctg
     2641 gtttaaaccc tgcagggggc agagcgcaca tcgcccacag tccccgagaa gttgggggga
     2701 ggggtcggca attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat
     2761 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta
     2821 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagcg ccggtgctaa
     2881 ctcgaacgct agctgtgcga tcgtttgcca ccatggcttt gaactcaggg tcaccaccag
     2941 ctattggacc ttactatgaa aaccatggat accaaccgga aaacccctat cccgcacagc
     3001 ccactgtggt ccccactgtc tacgaggtgc atccggctca gtactacccg tcccccgtgc
     3061 cccagtacgc cccgagggtc ctgacgcagg cttccaaccc cgtcgtctgc acgcagccca
     3121 aatccccatc cgggacagtg tgcacctcaa agactaagaa agcactgtgc atcaccttga
     3181 ccctggggac cttcctcgtg ggagctgcgc tggccgctgg cctactctgg aagttcatgg
     3241 gcagcaagtg ctccaactct gggatagagt gcgactcctc aggtacctgc atcaacccct
     3301 ctaactggtg tgatggcgtg tcacactgcc ccggcgggga ggacgagaat cggtgtgttc
     3361 gcctctacgg accaaacttc atccttcagg tgtactcatc tcagaggaag tcctggcacc
     3421 ctgtgtgcca agacgactgg aacgagaact acgggcgggc ggcctgcagg gacatgggct
     3481 ataagaataa tttttactct agccaaggaa tagtggatga cagcggatcc accagcttta
     3541 tgaaactgaa cacaagtgcc ggcaatgtcg atatctataa aaaactgtac cacagtgatg
     3601 cctgttcttc aaaagcagtg gtttctttac gctgtatagc ctgcggggtc aacttgaact
     3661 caagccgcca gagcaggatc gtgggcggcg agagcgcgct cccgggggcc tggccctggc
     3721 aggtcagcct gcacgtccag aacgtccacg tgtgcggagg ctccatcatc acccccgagt
     3781 ggatcgtgac agccgcccac tgcgtggaaa aacctcttaa caatccatgg cattggacgg
     3841 catttgcggg gattttgaga caatctttca tgttctatgg agccggatac caagtagaaa
     3901 aagtgatttc tcatccaaat tatgactcca agaccaagaa caatgacatt gcgctgatga
     3961 agctgcagaa gcctctgact ttcaacgacc tagtgaaacc agtgtgtctg cccaacccag
     4021 gcatgatgct gcagccagaa cagctctgct ggatttccgg gtggggggcc accgaggaga
     4081 aagggaagac ctcagaagtg ctgaacgctg ccaaggtgct tctcattgag acacagagat
     4141 gcaacagcag atatgtctat gacaacctga tcacaccagc catgatctgt gccggcttcc
     4201 tgcaggggaa cgtcgattct tgccagggtg acagtggagg gcctctggtc acttcgaaga
     4261 acaatatctg gtggctgata ggggatacaa gctggggttc tggctgtgcc aaagcttaca
     4321 gaccaggagt gtacgggaat gtgatggtat tcacggactg gatttatcga caaatgaggg
     4381 cagacggcta gaacccagct ttgttgtaca aaccgctgtt cctaggaatc ccgaggcctc
     4441 taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg
     4501 ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt
     4561 aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta
     4621 gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta
     4681 gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc
     4741 ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga
     4801 aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg
     4861 tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc
     4921 tcttcctcat ctccgggcct ttcggacccg acattagcgc tacagcttaa gcgggccacc
     4981 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     5041 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     5101 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     5161 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     5221 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     5281 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     5341 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     5401 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     5461 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     5521 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     5581 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     5641 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5701 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5761 gacgagttct tctgaggatc cgtttaaacg ttgttgtttg gggttgaatt actcttctag
     5821 accgcggcgc gcccgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     5881 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     5941 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     6001 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     6061 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     6121 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     6181 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     6241 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     6301 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     6361 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     6421 ccccgcatcg ataccgtcga cctcgatcga gacctagaaa aacatggagc aatcacaagt
     6481 agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag
     6541 gtgggttttc cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta
     6601 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     6661 agacaagata tccttgatct gtggatctac cacacacaag gctacttccc tgattggcag
     6721 aactacacac cagggccagg gatcagatat ccactgacct ttggatggtg ctacaagcta
     6781 gtaccagttg agcaagagaa ggtagaagaa gccaatgaag gagagaacac ccgcttgtta
     6841 caccctgtga gcctgcatgg gatggatgac ccggagagag aagtattaga gtggaggttt
     6901 gacagccgcc tagcatttca tcacatggcc cgagagctgc atccggactg tactgggtct
     6961 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     7021 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     7081 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagcatg
     7141 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     7201 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     7261 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     7321 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     7381 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     7441 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     7501 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     7561 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     7621 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     7681 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     7741 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     7801 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     7861 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     7921 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     7981 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     8041 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     8101 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     8161 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     8221 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     8281 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     8341 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     8401 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     8461 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     8521 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     8581 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     8641 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     8701 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     8761 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     8821 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     8881 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     8941 ccacctgac
//
