LOCUS       sequence-348086-        7057 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  knockout mouse Olfr2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7057)
  AUTHORS   Wu L, Hollinshead KER, Hao Y, Au C, Kroehling L, Ng C, Lin WY, Li D,
            Silva HM, Shin J, Lafaille JJ, Possemato R, Pacold ME, 
            Papagiannakopoulos T, Kimmelman AC, Satija R, Littman DR
  TITLE     Niche-Selective Inhibition of Pathogenic Th17 Cells by Targeting 
            Metabolic Redundancy.
  JOURNAL   Cell. 2020 Aug 6;182(3):641-654.e20. doi: 
            10.1016/j.cell.2020.06.014. Epub 2020 Jul 1.
  PUBMED    32615085
REFERENCE   2  (bases 1 to 7057)
  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: 
            "10.1016/j.cell.2020.06.014"; journalName: "Cell"; date: "2020-08-6-
            6"; volume: "182"; issue: "3"; pages: "641-654.e20"
COMMENT     Sequence Label: pSIN-Olfr2
FEATURES             Location/Qualifiers
     source          1..7057
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(79..99)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        84..413
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      264..399
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     435..454
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(535..552)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(706..1294)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(786..805)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1465..2256,2257..2325))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2088..2107
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2326..2430)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2498..2516
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        2616..2995
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2996..3199
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3150..3170
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             3200..3375
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    3438..3637
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             3838..4253
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     4183..4205
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     4221..4237
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        4300..4548
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     4300..4320
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4471..4490
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        4570..4645
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4752..4963
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             4983..5468
                     /codon_start=1
                     /product="mouse GPI-anchored cell surface glycoprotein of 
                     the immunoglobulin superfamily"
                     /label=Thy1.1
                     /note="This allele of Thy1 is also known as CD90.1."
                     /translation="MNPAISVALLLSVLQVSRGQKVTSLTACLVNQNLRLDCRHENNTK
                     DNSIQHEFSLTREKRKHVLSGTLGIPEHTYRSRVTLSNQPYIKVLTLANFTTKDEGDYF
                     CELRVSGANPMSSNKSISVYRDKLVKCGGISLLVQNTSWMLLLLLSLSLLQALDFISL"
     CDS             5478..5534
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             5529..6329
                     /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="GPMIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5589..5608)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6199..6218
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             6450..6875
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(6978..7000)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg
       61 gagtgtagat ccggctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
      121 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
      181 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
      241 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
      301 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
      361 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctta
      421 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
      481 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct
      541 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
      601 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
      661 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
      721 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
      781 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
      841 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
      901 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
      961 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     1021 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     1081 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     1141 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     1201 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     1261 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     1321 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     1381 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     1441 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     1501 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     1561 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     1621 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     1681 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     1741 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg
     1801 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     1861 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     1921 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     1981 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     2041 atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc
     2101 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     2161 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     2221 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     2281 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     2341 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     2401 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     2461 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     2521 tcgtgttcaa gaattagctt ggccattgca tacgttgtat ccatatcata atatgtacat
     2581 ttatattggc tcatgtccaa cattaccgcc atgttgacat tgattattga ctagttatta
     2641 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     2701 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     2761 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     2821 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     2881 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     2941 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     3001 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     3061 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     3121 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     3181 ggtctatata agcagagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
     3241 tccgatagac tgcgtcgccc gggtacccgt attcccaata aagcctcttg ctgtttgcat
     3301 ccgaatcgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccacga
     3361 cgggggtctt tcatttgggg gctcgtccgg gatttggaga cccctgccca gggaccaccg
     3421 acccaccacc gggaggtaag ctggccagca acttatctgt gtctgtccga ttgtctagtg
     3481 tctatgtttg atgttatgcg cctgcgtctg tactagttag ctaactagct ctgtatctgg
     3541 cggacccgtg gtggaactga cgagttctga acacccggcc gcaaccctgg gagacgtccc
     3601 agggactttg ggggccgttt ttgtggcccg acctgaggaa gggagtcgat gtggaatccg
     3661 accccgtcag gatatgtggt tctggtagga gacgagaacc taaaacagtt cccgcctccg
     3721 tctgaatttt tgctttcggt ttggaaccga agccgcgcgt cttgtctgct gcagcgctgc
     3781 agcatcgttc tgtgttgtct ctgtctgact gtgtttctgt atttgtctga aaattagggc
     3841 cagactgtta ccactccctt aagtttgacc ttaggtcact ggaaagatgt cgagcggatc
     3901 gctcacaacc agtcggtaga tgtcaagaag agacgttggt taccttctgc tctgcagaat
     3961 ggccaacctt taacgtcgga tggccgcgag acggcacctt taaccgagac ctcatcaccc
     4021 aggttaagat caaggtcttt tcacctggcc cgcatggaca cccagaccag gtcccctaca
     4081 tcgtgacctg ggaagccttg gcttttgacc cccctccctg ggtcaagccc tttgtacacc
     4141 ctaagcctcc gcctcctctt cctccatccg ccccgtctct cccccttgaa cctcctcgtt
     4201 cgaccccgcc tcgatcctcc ctttatccag ccctcactcc ttctctaggc gccggaattg
     4261 aagatcttta attaacgatc tatcgattct agatctagag agggcctatt tcccatgatt
     4321 ccttcatatt tgcatatacg atacaaggct gttagagaga taattagaat taatttgact
     4381 gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag
     4441 tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag
     4501 tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccga aatagtagtg
     4561 cccgcagtgg ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact
     4621 tgaaaaagtg gcaccgagtc ggtgcttttt tgaatttaga tcttgagaca aggtgccggg
     4681 ccgggaattc ggcgaattgg gcccgacgtc gcatgctccc ggcccctcga gtggctccgg
     4741 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt
     4801 cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt
     4861 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc
     4921 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtgtcgt gacgcggcca
     4981 ccatgaaccc agccatcagc gtcgctctcc tgctctcagt cttgcaggtg tcccgagggc
     5041 agaaggtgac cagcctgaca gcctgcctgg tgaaccaaaa ccttcgcctg gactgccgcc
     5101 atgagaataa caccaaggat aactccatcc agcatgagtt cagcctgacc cgagagaaga
     5161 ggaagcacgt gctctcaggc accctcggga tacccgagca cacgtaccgc tcccgcgtca
     5221 ccctctccaa ccagccctat atcaaggtcc ttaccctagc caacttcacc accaaggatg
     5281 agggcgacta cttttgtgag cttcgagtct cgggcgcgaa tcccatgagc tccaataaaa
     5341 gtatcagtgt gtatagagac aaactggtca agtgtggcgg cataagcctg ctggttcaga
     5401 acacatcctg gatgctgctg ctgctgcttt ccctctccct cctccaagcc ctggacttca
     5461 tttctctggg cagcggcgcc accaacttca gcctgctgaa gcaggccggc gacgtggagg
     5521 agaaccccgg ccccatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     5581 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     5641 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     5701 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     5761 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     5821 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     5881 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     5941 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     6001 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     6061 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     6121 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     6181 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     6241 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     6301 tctatcgcct tcttgacgag ttcttctgat tagatatcat cagtggtcca ggctctagtt
     6361 ttgactcaac aatatcacca gctgaagcct atagagtacg agccatagat aaaataaaag
     6421 attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag
     6481 ctagcttaag taacgccatt ttgcaaggca tggaaaaata cataactgag aatagagaag
     6541 ttcagatcaa ggtcaggaac agatggaaca gggtcgaccc tagagaacca tcagatgttt
     6601 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
     6661 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
     6721 ctcactcggg gcgccagtcc tccgattgac tgagtcgccc gggtacccgt gtatccaata
     6781 aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg
     6841 agtgattgac tacccgtcag cgggggtctt tcatttgggg gctcgtccgg gatcgggaga
     6901 cccctgccca gggaccaccg acccaccacc gggaggtaag ctggctgcct cgcgcgtttc
     6961 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     7021 taagcggatg ccgggagcag acaagcccgt cagggcg
//
