LOCUS       sequence-304631-        8932 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing a miR-146a decoy sponge.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8932)
  AUTHORS   Emming S, Bianchi N, Polletti S, Balestrieri C, Leoni C, Montagner 
            S, Chirichella M, Delaleu N, Natoli G, Monticelli S
  TITLE     A molecular network regulating the proinflammatory phenotype of 
            human memory T lymphocytes.
  JOURNAL   Nat Immunol. 2020 Apr;21(4):388-399. doi: 10.1038/s41590-020-0622-8.
            Epub 2020 Mar 16.
  PUBMED    32205878
REFERENCE   2  (bases 1 to 8932)
  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"; doi: 
            "10.1038/s41590-020-0622-8"; journalName: "Nat Immunol"; date: 
            "2020-04"; volume: "21"; issue: "4"; pages: "388-399"
COMMENT     Sequence Label: pSCALPS_miR-146aSponge_mCherry_GFP
FEATURES             Location/Qualifiers
     source          1..8932
                     /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"
     primer_bind     complement(240..260)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        245..574
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      425..560
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     487..506
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        586..965
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        967..1165
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1120..1140
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             1183..1363
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1410..1535
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2032..2265
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2449..2493
                     /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    2791..2908
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3045..3452
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      3487..3496
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3493..4203
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3638..3656)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3906..3925
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     regulatory      5112..5121
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5118..5813
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     misc_feature    5823..6411
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5876..5896)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6294..6305)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6929..7109
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7180..7768)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7260..7279)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7939..8730,8731..8799))
                     /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     8562..8581
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8800..8904)
                     /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 acgcgctgtg
      241 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
      301 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
      361 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
      421 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
      481 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
      541 aggaggcttt tttggaggcc taggcttttg caaaaagctc gcgttgacat tgattattga
      601 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
      661 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      721 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      781 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      841 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      901 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      961 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     1021 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     1081 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     1141 tacggtggga ggtctatata agcagcgcgt tttgcctgta ctgggtctct ctggttagac
     1201 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     1261 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     1321 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg
     1381 acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag
     1441 cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg
     1501 gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat
     1561 cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata
     1621 tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat
     1681 cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag
     1741 aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga
     1801 taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca
     1861 ccgcacagca agcggccggc cgctgatctt cagacctgga ggaggagata tgagggacaa
     1921 ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc
     1981 caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt
     2041 gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac
     2101 ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc
     2161 tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc
     2221 aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg
     2281 ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc
     2341 tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaat taacaattac
     2401 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
     2461 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
     2521 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
     2581 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
     2641 acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga
     2701 gagagagaca gagacagatc cattcgatta gtgaacggat cggcactgcg tgcgccaatt
     2761 ctgcagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta
     2821 cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca
     2881 aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt
     2941 ttggttaatt aactgcagcc ccgataaaat aaaagatttt atttagtctc cagaaaaagg
     3001 ggggaatgaa agaccccacc tgtaggtttg gcaagctagc tgcagtaacg ccattttgca
     3061 aggcatggaa aaataccaaa ccaagaatag agaagttcag atcaagggcg ggtacatgaa
     3121 aatagctaac gttgggccaa acaggatatc tgcggtgagc agtttcggcc ccggcccggg
     3181 gccaagaaca gatggtcacc gcagtttcgg ccccggcccg aggccaagaa cagatggtcc
     3241 ccagatatgg cccaaccctc agcagtttct taagacccat cagatgtttc caggctcccc
     3301 caaggacctg aaatgaccct gcgccttatt tgaattaacc aatcagcctg cttctcgctt
     3361 ctgttcgcgc gcttctgctt cccgagctct ataaaagagc tcacaacccc tcactcggcg
     3421 cgccagtcct ccgacagact gagtcgcccg ggggtctaga agcgctggat ccgtttaaac
     3481 gcggccgcca ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
     3541 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
     3601 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
     3661 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
     3721 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
     3781 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
     3841 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     3901 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     3961 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     4021 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     4081 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     4141 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     4201 taagaattcg tcgcccgggg gtctagaagc aacccatgga cagagttctc accgtaaccc
     4261 atggacagag ttctcacctt aacccatgga cagagttctc accgtaaccc atggacagag
     4321 ttctcaccgt aacccatgga cagagttctc accgtaaccc atggacagag ttctcaccgt
     4381 aacccatgga cagagttctc accgtaaccc atggacagag ttctcaccgt aacccatgga
     4441 cagagttctc accgtaaccc atggacagag ttctcaccgt aacccatgga cagagttctc
     4501 accgtaaccc atggacagag ttctcaccgt aacccatgga cagagttctc accgtaaccc
     4561 atggacagag ttctcaccgt aacccatgga caggctggat ccgtttaaac gcgggaattc
     4621 tcgaccttga ctggcggcgc gaccttgagg cctgcgttcg cctcagttgc cccctctgtg
     4681 caatggggag acgcgcctca tcgcttgaca acggccgaag agccgccgcg cttccgtctc
     4741 ccgcgtgcgc gcgccatgct gcccaccccc gttccgcact gaccctcccc cgtgccccgc
     4801 gtcccgtact gccgccccgc cccgagtccc atgccgcagc caccgcgacg gagcccgcag
     4861 gcgggaacct gcctccgcgc gttagcgcgc acgcgcgcct catgtgtcgt ccccatcagc
     4921 gccggcttcc gtctataggc cagatgcact gtcactctgg cgaagtcgca gacccgattg
     4981 gccgggacgg aggcgcgaga ccgggttgcg ggcggggccg aacgtggtat aaaaggggcg
     5041 ggaggccagg ctcgtgccgt tttgcagacg ccaccgccga ggaaaaccgt gtactattag
     5101 ccacgcgtag agccaccatg gcccagtcca agcacggcct gaccaaggag atgaccatga
     5161 agtaccgcat ggagggctgc gtggacggcc acaagttcgt gatcaccggc gagggcatcg
     5221 gctacccctt caagggcaag caggccatca acctgtgcgt ggtggagggc ggccccttgc
     5281 ccttcgccga ggacatcttg tccgccgcct tcatgtacgg caaccgcgtg ttcaccgagt
     5341 acccccagga catcgtcgac tacttcaaga actcctgccc cgccggctac acctgggacc
     5401 gctccttcct gttcgaggac ggcgccgtgt gcatctgcaa cgccgacatc accgtgagcg
     5461 tggaggagaa ctgcatgtac cacgagtcca agttctacgg cgtgaacttc cccgccgacg
     5521 gccccgtgat gaagaagatg accgacaact gggagccctc ctgcgagaag atcatccccg
     5581 tgcccaagca gggcatcttg aagggcgacg tgagcatgta cctgctgctg aaggacggtg
     5641 gccgcttgcg ctgccagttc gacaccgtgt acaaggccaa gtccgtgccc cgcaagatgc
     5701 ccgactggca cttcatccag cacaagctga cccgcgagga ccgcagcgac gccaagaacc
     5761 agaagtggca cctgaccgag cacgccatcg cctccggctc cgccttgccc tgagcgatcg
     5821 ctaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     5881 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     5941 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     6001 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     6061 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     6121 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     6181 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc
     6241 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     6301 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     6361 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cttaatcgcg
     6421 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga
     6481 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt
     6541 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     6601 ggggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat
     6661 ctaccacaca caaggctact tccctgattg gcagaactac acaccagggc cagggatcag
     6721 atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag agaaggtaga
     6781 agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc atgggatgga
     6841 tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat ttcatcacat
     6901 ggcccgagag ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg
     6961 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     7021 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     7081 cttttagtca gtgtggaaaa tctctagcag ggcccgtttc atgtgagcaa aaggccagca
     7141 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     7201 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     7261 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     7321 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     7381 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     7441 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     7501 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     7561 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7621 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7681 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7741 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7801 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7861 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7921 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7981 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     8041 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     8101 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     8161 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     8221 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     8281 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     8341 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     8401 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     8461 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     8521 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     8581 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     8641 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8701 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8761 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8821 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8881 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg ac
//
