LOCUS       sequence-334962-        8963 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TMPRSS2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8963)
  AUTHORS   Yurkovetskiy L, Wang X, Pascal KE, Tomkins-Tinch C, Nyalile TP, Wang
            Y, Baum A, Diehl WE, Dauphin A, Carbone C, Veinotte K, Egri SB, 
            Schaffner SF, Lemieux JE, Munro JB, Rafique A, Barve A, Sabeti PC, 
            Kyratsous CA, Dudkina NV, Shen K, Luban J
  TITLE     Structural and Functional Analysis of the D614G SARS-CoV-2 Spike 
            Protein Variant.
  JOURNAL   Cell. 2020 Sep 15. pii: S0092-8674(20)31229-0. doi: 
            10.1016/j.cell.2020.09.032.
  PUBMED    32991842
REFERENCE   2  (bases 1 to 8963)
  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: "Cell. 2020 
            Sep 15. pii: S0092-8674(20)31229-0. doi: 
            10.1016/j.cell.2020.09.032."
COMMENT     Sequence Label: pscALPSblasti-TMPRSS2 Blasti
FEATURES             Location/Qualifiers
     source          1..8963
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    162..279
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        416..823
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      832..841
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2811..2820
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3001..3012
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     misc_feature    3225..3813
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3278..3298)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3696..3707)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4331..4511
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4582..5170)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4662..4681)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5341..6132,6133..6201))
                     /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     5964..5983
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6202..6306)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6381..6400)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6574..6594)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6579..6908
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6759..6894
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6821..6840
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        6920..7299
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7301..7499
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7454..7474
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             7517..7697
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7744..7869
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8366..8599
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8783..8827
                     /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"
ORIGIN
        1 tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag
       61 aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga tcggcactgc
      121 gtgcgccaat tctgcagaca aatggcagta ttcatccaca attttaaaag aaaagggggg
      181 attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact
      241 aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc
      301 agagatccag tttggttaat taactgcagc cccgataaaa taaaagattt tatttagtct
      361 ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag ctgcagtaac
      421 gccattttgc aaggcatgga aaaataccaa accaagaata gagaagttca gatcaagggc
      481 gggtacatga aaatagctaa cgttgggcca aacaggatat ctgcggtgag cagtttcggc
      541 cccggcccgg ggccaagaac agatggtcac cgcagtttcg gccccggccc gaggccaaga
      601 acagatggtc cccagatatg gcccaaccct cagcagtttc ttaagaccca tcagatgttt
      661 ccaggctccc ccaaggacct gaaatgaccc tgcgccttat ttgaattaac caatcagcct
      721 gcttctcgct tctgttcgcg cgcttctgct tcccgagctc tataaaagag ctcacaaccc
      781 ctcactcggc gcgccagtcc tccgacagac tgagtcgccc gggggtctag agccaccatg
      841 gctttgaact cagggtcacc accagctatt ggaccttact atgaaaacca tggataccaa
      901 ccggaaaacc cctatcccgc acagcccact gtggtcccca ctgtctacga ggtgcatccg
      961 gctcagtact acccgtcccc cgtgccccag tacgccccga gggtcctgac gcaggcttcc
     1021 aaccccgtcg tctgcacgca gcccaaatcc ccatccggga cagtgtgcac ctcaaagact
     1081 aagaaagcac tgtgcatcac cttgaccctg gggaccttcc tcgtgggagc tgcgctggcc
     1141 gctggcctac tctggaagtt catgggcagc aagtgctcca actctgggat agagtgcgac
     1201 tcctcaggta cctgcatcaa cccctctaac tggtgtgatg gcgtgtcaca ctgccccggc
     1261 ggggaggacg agaatcggtg tgttcgcctc tacggaccaa acttcatcct tcaggtgtac
     1321 tcatctcaga ggaagtcctg gcaccctgtg tgccaagacg actggaacga gaactacggg
     1381 cgggcggcct gcagggacat gggctataag aataattttt actctagcca aggaatagtg
     1441 gatgacagcg gatccaccag ctttatgaaa ctgaacacaa gtgccggcaa tgtcgatatc
     1501 tataaaaaac tgtaccacag tgatgcctgt tcttcaaaag cagtggtttc tttacgctgt
     1561 atagcctgcg gggtcaactt gaactcaagc cgccagagca ggatcgtggg cggcgagagc
     1621 gcgctcccgg gggcctggcc ctggcaggtc agcctgcacg tccagaacgt ccacgtgtgc
     1681 ggaggctcca tcatcacccc cgagtggatc gtgacagccg cccactgcgt ggaaaaacct
     1741 cttaacaatc catggcattg gacggcattt gcggggattt tgagacaatc tttcatgttc
     1801 tatggagccg gataccaagt agaaaaagtg atttctcatc caaattatga ctccaagacc
     1861 aagaacaatg acattgcgct gatgaagctg cagaagcctc tgactttcaa cgacctagtg
     1921 aaaccagtgt gtctgcccaa cccaggcatg atgctgcagc cagaacagct ctgctggatt
     1981 tccgggtggg gggccaccga ggagaaaggg aagacctcag aagtgctgaa cgctgccaag
     2041 gtgcttctca ttgagacaca gagatgcaac agcagatatg tctatgacaa cctgatcaca
     2101 ccagccatga tctgtgccgg cttcctgcag gggaacgtcg attcttgcca gggtgacagt
     2161 ggagggcctc tggtcacttc gaagaacaat atctggtggc tgatagggga tacaagctgg
     2221 ggttctggct gtgccaaagc ttacagacca ggagtgtacg ggaatgtgat ggtattcacg
     2281 gactggattt atcgacaaat gagggcagac ggctaggaat tctcgacctt gactggcggc
     2341 gcgaccttga ggcctgcgtt cgcctcagtt gccccctctg tgcaatgggg agacgcgcct
     2401 catcgcttga caacggccga agagccgccg cgcttccgtc tcccgcgtgc gcgcgccatg
     2461 ctgcccaccc ccgttccgca ctgaccctcc cccgtgcccc gcgtcccgta ctgccgcccc
     2521 gccccgagtc ccatgccgca gccaccgcga cggagcccgc aggcgggaac ctgcctccgc
     2581 gcgttagcgc gcacgcgcgc ctcatgtgtc gtccccatca gcgccggctt ccgtctatag
     2641 gccagatgca ctgtcactct ggcgaagtcg cagacccgat tggccgggac ggaggcgcga
     2701 gaccgggttg cgggcggggc cgaacgtggt ataaaagggg cgggaggcca ggctcgtgcc
     2761 gttttgcaga cgccaccgcc gaggaaaacc gtgtactatt agccacgcgt gccgccatgg
     2821 ctaaaccact gtcccaggaa gaatccacac tgatcgaacg agcaaccgct actatcaact
     2881 caatcccaat ctccgaagac tactctgtgg cctccgccgc cctgagcagc gacggacgaa
     2941 tcttcaccgg cgtgaacgtc taccacttta ccggcggccc ctgcgctgag ctggtggtcc
     3001 tgggaactgc agccgctgca gccgctggaa atctgacctg tatcgtggct atcgggaacg
     3061 aaaatagggg aatcctgtct ccatgcggcc ggtgtagaca ggtcctgctg gatctgcatc
     3121 ccgggatcaa ggcaattgtg aaagactcag atggacagcc cacagccgtc ggcattcgcg
     3181 agctgctgcc tagcggctat gtgtgggaag ggtgagcgat cgctaatcaa cctctggatt
     3241 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     3301 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     3361 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     3421 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     3481 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     3541 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     3601 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct
     3661 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     3721 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     3781 cgagtcggat ctccctttgg gccgcctccc cgcttaatcg cgtcgagacc tagaaaaaca
     3841 tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc
     3901 acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac
     3961 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     4021 aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta
     4081 cttccctgat tggcagaact acacaccagg gccagggatc agatatccac tgacctttgg
     4141 atggtgctac aagctagtac cagttgagca agagaaggta gaagaagcca atgaaggaga
     4201 gaacacccgc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt
     4261 attagagtgg aggtttgaca gccgcctagc atttcatcac atggcccgag agctgcatcc
     4321 ggactgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     4381 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     4441 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     4501 aatctctagc agggcccgtt tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4561 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4621 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4681 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4741 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4801 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4861 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4921 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4981 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5041 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5101 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5161 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5221 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5281 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5341 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5401 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5461 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5521 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5581 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5641 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5701 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5761 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5821 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5881 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5941 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6001 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6061 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6121 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6181 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6241 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6301 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     6361 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     6421 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     6481 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
     6541 cttcgcgatg tacgggccag atatacgcgc tgtggaatgt gtgtcagtta gggtgtggaa
     6601 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     6661 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     6721 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     6781 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     6841 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     6901 tttgcaaaaa gctcgcgttg acattgatta ttgactagtt attaatagta atcaattacg
     6961 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     7021 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     7081 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     7141 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     7201 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     7261 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     7321 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     7381 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     7441 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcagc
     7501 gcgttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     7561 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     7621 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     7681 gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag
     7741 gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg
     7801 cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc
     7861 gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg
     7921 ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa
     7981 cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga
     8041 cagctacaac catcccttca gacaggatca gaagaactta gatcattata taatacagta
     8101 gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac
     8161 aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc cggccgctga
     8221 tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata
     8281 aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc
     8341 agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag
     8401 gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg
     8461 gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc
     8521 aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc
     8581 taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg
     8641 ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga
     8701 cctggatgga gtgggacaga gaattaacaa ttacacaagc ttaatacact ccttaattga
     8761 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
     8821 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
     8881 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
     8941 agttaggcag ggatattcac cat
//
