LOCUS       sequence-363827-        4850 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Allows for constituitive mammalian expression of the MERS-CoV 3CL 
            protease via transfection or lentivirus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4850)
  AUTHORS   Resnick SJ, Iketani S, Hong SJ, Zask A, Liu H, Kim S, Melore S, Lin 
            FY, Nair MS, Huang Y, Lee S, Tay NES, Rovis T, Yang HW, Xing L, 
            Stockwell BR, Ho DD, Chavez A
  TITLE     Inhibitors of Coronavirus 3CL Proteases Protect Cells from 
            Protease-Mediated Cytotoxicity.
  JOURNAL   J Virol. 2021 Jun 24;95(14):e0237420. doi: 10.1128/JVI.02374-20. 
            Epub 2021 Jun 24.
  PUBMED    33910954
REFERENCE   2  (bases 1 to 4850)
  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 Virol."; 
            date: "2021-06-24"; volume: "95(14):e0237420. doi"; pages: " 
            10.1128/JVI.02374-20. Epub 2021 Jun 24"
COMMENT     Sequence Label: pLEX307-MERS-CoV-3CL
FEATURES             Location/Qualifiers
     source          1..4850
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    93..114
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(129..134,135..152,153..159)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    167..183
                     /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     172..194
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     191..207
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     191..207
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     226..246
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        228..246
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        274..500
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             501..681
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    728..853
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1346..1579
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1764..1808
                     /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"
     rep_origin      2108..2243
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2263..2282)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2264..2282)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2292..2309)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2292..2308)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2301..2323)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2450..2905
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2537..2556)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2747..2768
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2931..3035
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3036..3104,3105..3896)
                     /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     complement(3254..3273)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4067..4655
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4556..4575
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4809..4826
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
       61 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
      121 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
      181 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac
      241 taaagggaac aaaagctgga gctgcaagct taatgtagtc ttatgcaata ctcttgtagt
      301 cttgcaacat ggtaacgatg agttagcaac atgccttaca aggagagaaa aagcaccgtg
      361 catgccgatt ggtggaagta aggtggtacg atcgtgcctt attaggaagg caacagacgg
      421 gtctgacatg gattggacga accactgaat tgccgcattg cagagatatt gtatttaagt
      481 gcctagctcg atacataaac gggtctctct ggttagacca gatctgagcc tgggagctct
      541 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
      601 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
      661 cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc
      721 agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg
      781 gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg
      841 gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt
      901 aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct
      961 agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact
     1021 gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac
     1081 agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt
     1141 agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga
     1201 tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata
     1261 aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc
     1321 agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag
     1381 gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg
     1441 gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc
     1501 aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc
     1561 taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg
     1621 ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga
     1681 cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg
     1741 aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg
     1801 caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa
     1861 tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata
     1921 gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac
     1981 ccatgcattg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     2041 aggcagaagt atgcaaagca tgcgtctcaa ttagtcagca accatagtcc cgcccctaac
     2101 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     2161 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     2221 gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat
     2281 tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     2341 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     2401 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
     2461 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2521 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2581 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     2641 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     2701 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     2761 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     2821 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     2881 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     2941 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3001 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     3061 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     3121 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     3181 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     3241 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     3301 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     3361 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3421 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3481 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3541 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     3601 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     3661 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     3721 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     3781 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     3841 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     3901 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     3961 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4021 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4081 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4141 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4201 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4261 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4321 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4381 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4441 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4501 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4561 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4621 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4681 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     4741 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     4801 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
//
