LOCUS       sequence-297111-        8943 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector genome to ectopically express human Ace2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8943)
  AUTHORS   Rebendenne A, Valadao ALC, Tauziet M, Maarifi G, Bonaventure B, 
            McKellar J, Planes R, Nisole S, Arnaud-Arnould M, Moncorge O, Goujon
            C
  TITLE     SARS-CoV-2 triggers an MDA-5-dependent interferon response which is 
            unable to control replication in lung epithelial cells.
  JOURNAL   J Virol. 2021 Jan 29. pii: JVI.02415-20. doi: 10.1128/JVI.02415-20.
  PUBMED    33514628
REFERENCE   2  (bases 1 to 8943)
  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. 
            2021 Jan 29. pii: JVI.02415-20. doi: 10.1128/JVI.02415-20."
COMMENT     Sequence Label: RRL.sin.cPPT.SFFV/Ace2.WPRE (MT136)
FEATURES             Location/Qualifiers
     source          1..8943
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        26..130
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(131..199,200..991)
                     /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(349..368)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1162..1750
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1651..1670
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1904..1921
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2038..2059
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2074..2079,2080..2097,2098..2104)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2112..2128
                     /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     2117..2139
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2136..2152
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2136..2152
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2171..2191
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2173..2191
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2219..2445
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2446..2626
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2673..2798
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3291..3524
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3709..3753
                     /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    4020..4137
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4184..4591
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     misc_feature    7026..7614
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7079..7099)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7497..7508)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7701..7934
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8006..8127
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8043..8062)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8097..8116
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8146..8281
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8301..8320)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8302..8320)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8330..8347)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8330..8346)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8339..8361)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8488..8916
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8575..8594)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8785..8806
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
       61 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
      121 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
      181 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
      241 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
      301 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
      361 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
      421 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
      481 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
      541 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
      601 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
      661 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
      721 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
      781 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
      841 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
      901 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
      961 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     1021 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     1081 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     1141 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     1201 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     1261 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     1321 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     1381 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     1441 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     1501 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     1561 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     1621 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     1681 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     1741 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     1801 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     1861 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     1921 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     1981 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     2041 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     2101 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     2161 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg
     2221 tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc
     2281 ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt
     2341 gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg
     2401 cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta
     2461 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     2521 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     2581 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
     2641 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
     2701 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     2761 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     2821 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     2881 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
     2941 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     3001 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     3061 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     3121 ccaccgcaca gcaaggggcc gctgatcttc agacctggag gaggagatat gagggacaat
     3181 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     3241 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     3301 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     3361 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     3421 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     3481 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     3541 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     3601 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
     3661 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
     3721 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
     3781 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
     3841 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
     3901 acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga
     3961 gagagagaca gagacagatc cattcgatta gtgaacggat ctcgacggta tcggttaact
     4021 tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata
     4081 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttatc
     4141 gataatgaaa gaccccacct gtaggtttgg caagctagct gcagtaacgc cattttgcaa
     4201 ggcatggaaa aataccaaac caagaataga gaagttcaga tcaagggcgg gtacatgaaa
     4261 atagctaacg ttgggccaaa caggatatct gcggtgagca gtttcggccc cggcccgggg
     4321 ccaagaacag atggtcaccg cagtttcggc cccggcccga ggccaagaac agatggtccc
     4381 cagatatggc ccaaccctca gcagtttctt aagacccatc agatgtttcc aggctccccc
     4441 aaggacctga aatgaccctg cgccttattt gaattaacca atcagcctgc ttctcgcttc
     4501 tgttcgcgcg cttctgcttc ccgagctcta taaaagagct cacaacccct cactcggcgc
     4561 gccagtcctc cgacagactg agtcgcccgg ggggcggccg catgtcaagc tcttcctggc
     4621 tccttctcag ccttgttgct gtaactgctg ctcagtccac cattgaggaa caggccaaga
     4681 catttttgga caagtttaac cacgaagccg aagacctgtt ctatcaaagt tcacttgctt
     4741 cttggaatta taacaccaat attactgaag agaatgtcca aaacatgaat aatgctgggg
     4801 acaaatggtc tgccttttta aaggaacagt ccacacttgc ccaaatgtat ccactacaag
     4861 aaattcagaa tctcacagtc aagcttcagc tgcaggctct tcagcaaaat gggtcttcag
     4921 tgctctcaga agacaagagc aaacggttga acacaattct aaatacaatg agcaccatct
     4981 acagtactgg aaaagtttgt aacccagata atccacaaga atgcttatta cttgaaccag
     5041 gtttgaatga aataatggca aacagtttag actacaatga gaggctctgg gcttgggaaa
     5101 gctggagatc tgaggtcggc aagcagctga ggccattata tgaagagtat gtggtcttga
     5161 aaaatgagat ggcaagagca aatcattatg aggactatgg ggattattgg agaggagact
     5221 atgaagtaaa tggggtagat ggctatgact acagccgcgg ccagttgatt gaagatgtgg
     5281 aacatacctt tgaagagatt aaaccattat atgaacatct tcatgcctat gtgagggcaa
     5341 agttgatgaa tgcctatcct tcctatatca gtccaattgg atgcctccct gctcatttgc
     5401 ttggtgatat gtggggtaga ttttggacaa atctgtactc tttgacagtt ccctttggac
     5461 agaaaccaaa catagatgtt actgatgcaa tggtggacca ggcctgggat gcacagagaa
     5521 tattcaagga ggccgagaag ttctttgtat ctgttggtct tcctaatatg actcaaggat
     5581 tctgggaaaa ttccatgcta acggacccag gaaatgttca gaaagcagtc tgccatccca
     5641 cagcttggga cctggggaag ggcgacttca ggatccttat gtgcacaaag gtgacaatgg
     5701 acgacttcct gacagctcat catgagatgg ggcatatcca gtatgatatg gcatatgctg
     5761 cacaaccttt tctgctaaga aatggagcta atgaaggatt ccatgaagct gttggggaaa
     5821 tcatgtcact ttctgcagcc acacctaagc atttaaaatc cattggtctt ctgtcacccg
     5881 attttcaaga agacaatgaa acagaaataa acttcctgct caaacaagca ctcacgattg
     5941 ttgggactct gccatttact tacatgttag agaagtggag gtggatggtc tttaaagggg
     6001 aaattcccaa agaccagtgg atgaaaaagt ggtgggagat gaagcgagag atagttgggg
     6061 tggtggaacc tgtgccccat gatgaaacat actgtgaccc cgcatctctg ttccatgttt
     6121 ctaatgatta ctcattcatt cgatattaca caaggaccct ttaccaattc cagtttcaag
     6181 aagcactttg tcaagcagct aaacatgaag gccctctgca caaatgtgac atctcaaact
     6241 ctacagaagc tggacagaaa ctgttcaata tgctgaggct tggaaaatca gaaccctgga
     6301 ccctagcatt ggaaaatgtt gtaggagcaa agaacatgaa tgtaaggcca ctgctcaact
     6361 actttgagcc cttatttacc tggctgaaag accagaacaa gaattctttt gtgggatgga
     6421 gtaccgactg gagtccatat gcagaccaaa gcatcaaagt gaggataagc ctaaaatcag
     6481 ctcttggaga taaagcatat gaatggaacg acaatgaaat gtacctgttc cgatcatctg
     6541 ttgcatatgc tatgaggcag tactttttaa aagtaaaaaa tcagatgatt ctttttgggg
     6601 aggaggatgt gcgagtggct aatttgaaac caagaatctc ctttaatttc tttgtcactg
     6661 cacctaaaaa tgtgtctgat atcattccta gaactgaagt tgaaaaggcc atcaggatgt
     6721 cccggagccg tatcaatgat gctttccgtc tgaatgacaa cagcctagag tttctgggga
     6781 tacagccaac acttggacct cctaaccagc cccctgtttc catatggctg attgtttttg
     6841 gagttgtgat gggagtgata gtggttggca ttgtcatcct gatcttcact gggatcagag
     6901 atcggaagaa gaaaaataaa gcaagaagtg gagaaaatcc ttatgcctcc atcgatatta
     6961 gcaaaggaga aaataatcca ggattccaaa acactgatga tgttcagacc tccttttagc
     7021 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     7081 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     7141 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     7201 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     7261 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     7321 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     7381 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
     7441 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     7501 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     7561 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa
     7621 ttcgagctcg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     7681 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct
     7741 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     7801 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     7861 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     7921 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa
     7981 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata
     8041 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     8101 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact
     8161 ccgcccagty ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     8221 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     8281 ctagggacgt acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg
     8341 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     8401 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     8461 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     8521 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     8581 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     8641 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     8701 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     8761 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     8821 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     8881 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     8941 ttt
//
