LOCUS       sequence-240347-        8249 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral overexpression of Lrrc40 - truncated version with 
            seemingly same DE genes in Th2 as MSCV-Lrrc40-IRES-BFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8249)
  AUTHORS   Henriksson J, Chen X, Gomes T, Ullah U, Meyer KB, Miragaia R, Duddy 
            G, Pramanik J, Yusa K, Lahesmaa R, Teichmann SA
  TITLE     Genome-wide CRISPR Screens in T Helper Cells Reveal Pervasive 
            Crosstalk between Activation and Differentiation.
  JOURNAL   Cell. 2019 Jan 7. pii: S0092-8674(18)31569-1. doi: 
            10.1016/j.cell.2018.11.044.
  PUBMED    30639098
REFERENCE   2  (bases 1 to 8249)
  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. 2019 
            Jan 7. pii: S0092-8674(18)31569-1. doi: 10.1016/j.cell.2018.11.044."
COMMENT     Sequence Label: MSCV-Lrrc40-trunc-IRES-BFP
FEATURES             Location/Qualifiers
     source          1..8249
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..517
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    581..922
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             989..1405
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1335..1357
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1373..1389
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    3212..3761
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3356..3373)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3583..3602
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3764..3766,3767..3769,3770..4483)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(3809..3830)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     LTR             4644..5158
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5320..5336
                     /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)."
     promoter        complement(join(5344..5350,5351..5368,5369..5374))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5389..5410
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5527..5544)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5698..6286)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5778..5797)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6457..7248,7249..7317))
                     /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     7080..7099
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7318..7422)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7490..7508
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7546..7568)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7668..7687
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7881..7903
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8031..8050
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
      121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
      241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
      301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
      361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
      421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
      481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
      541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
      601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
      661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
      721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
      781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
      841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
      901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
      961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
     1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
     1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
     1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
     1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
     1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
     1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
     1381 agccctcact ccttctctag gcgccggaat tcagatctta cgtagctagc ggatcttaac
     1441 catgtcgcgc cagctgagag cccctaggtt tgaccctcgt gcgggattcc acgcggaggg
     1501 caaggacaga ggcccctctg ttccccaggg cctgctgaag gcagcgagaa gcagcggcca
     1561 actcaacttg gcgggaagga acctcgggga agtccctcag tgtgtttgga gaataaatgt
     1621 ggacattcct gaagaggcta atcagaatct ttcattcagt tctactgaac gatggtggga
     1681 tcagacagat ctgaccaaac tcatcatctc cagcaataaa cttcagtctc tctctgatga
     1741 cctccgactc ttgcctgccc ttactgttct tgatccataa caaactgaaa atactgcctg
     1801 aagaaattac aagcttaaaa aacctgagga cgctgcacct ccagcacaat gagctgactt
     1861 gcatccccga gggatttgag catctttcct gcttagaaga tttagatctt tcaagcaatc
     1921 gtcttgcaac tgtgcctgct gattttgctt tgctgtccag tctgttgcga ctcaatcttt
     1981 ccagtaacca gctgaagaat ttgccagcag aaataagcag aatgaaaaga ttaaagcact
     2041 tggattgtga tgccaatctc ctggagactg tgcctcctga tgtgggtagc atggaatcac
     2101 tggaattgct ttatttgcgg aggaataaat tgcgtgttct acctgaattt ccttcttgta
     2161 ggcagctaaa ggaattacac ctagctgaaa accagataga gaagttaggg gcagagcatc
     2221 ttcagcacct gcaggccatc cttgtcctgg atctaagggg caataagctg cggtctgtcc
     2281 ctgaggaaat ggccctgctg cagtctttgg aaaggcttga cctgagcaac aatgatatta
     2341 gtagtctgcc ctgctcattg ggaaatcttc atttgaaatt cctggcacta gaaggaaatc
     2401 ctctaagaac cattcgaaga gaaattatag ctaaaggaac tcaagaagtt ctaaaatacc
     2461 tgcgcagcaa gatcaaagat gatagaacca atcaaaatga ttctgttcct gagactgcca
     2521 tgacactacc aagtgaagcc agagtcaata tccatgccat agccacacta aagctgctag
     2581 attatagtga taaacaagca accttgattc ctgatgacct atttgatgcc acaaaaacga
     2641 ccctcatcac ttccatcaac ttcagcaaaa atcagctgtg cgagattcca caaaggattg
     2701 ttgagctgaa ggaaatggtg ttggatatca acctcagttt taacaagctt tcatttatat
     2761 cccacgagtt atgcttgctt cagaaattga catttttaga tttaaggaat aattttttaa
     2821 gttctttgcc tgaagagatg tcatcactga caaaactgca aacgatcaat ctttccttta
     2881 acaggttcaa agtgtttcct gaagttctgt atcgaatctc cacactggaa gcggttctga
     2941 tcagtaacaa tcaggttggc tctgtggacc ctcagaagat gaagctgatg gaaaatctga
     3001 atactttaga ccttcaaaac aatgaccttt tgcagattcc cccagagctt gggaactgtg
     3061 tgcagctacg gacactactg ctagatggaa atccattccg agttcctcga gcggccatat
     3121 taatgaaagg cacagctgct gtgctggagt atttgagaga tcgaatccct gcctgatagg
     3181 ctcgagcggg atcaattccg ccccccccct aacgttactg gccgaagccg cttggaataa
     3241 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg
     3301 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc
     3361 gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct
     3421 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac
     3481 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc
     3541 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta
     3601 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg
     3661 cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg
     3721 aaccacgggg acgtggtttt cctttgaaaa acacgataat accatggtga gcaagggcga
     3781 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     3841 caagttcagc gtgaggggcg agggcgaggg cgatgccacc aacggcaagc tgaccctgaa
     3901 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgag
     3961 ccacggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
     4021 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcac
     4081 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     4141 gaagggcgtc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaactt
     4201 caacagccac aacatctata tcatggccgt caagcagaag aacggcatca aggtgaactt
     4261 caagatccgc cacaacgtgg aggacggcag cgtgcagctc gccgaccact accagcagaa
     4321 cacccccatc ggcgacggcc ccgtgctgct gcccgacagc cactacctga gcacccagtc
     4381 cgtgctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttccgcac
     4441 cgccgccggg atcactctcg gcatggacga gctgtacaag taagcggccg cgactctaga
     4501 gtcgacctgc aggcatgcaa gcttcaggta gccggctaac gttaacaacc ggtacctcta
     4561 gaactatagc tagcatgcgc aaatttaaag cgctgatatc gataaaataa aagattttat
     4621 ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt
     4681 aagtaacgcc attttgcaag gcatggaaaa tacataactg agaatagaga agttcagatc
     4741 aaggttagga acagagagac agcagaatat gggccaaaca ggatatctgt ggtaagcagt
     4801 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtcccg ccctcagcag
     4861 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
     4921 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
     4981 gctcaataaa agagcccaca acccctcact cggcgcgcca gtcctccgat agactgcgtc
     5041 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
     5101 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcatgg
     5161 gtaacagttt cttgaagttg gagaacaaca ttctgagggt aggagtcgaa tattaagtaa
     5221 tcctgactca attagccact gttttgaatc cacatactcc aatactcctg aaatagttca
     5281 ttatggacag cgcagaaaga gctggggaga attgtgaaat tgttatccgc tcacaattcc
     5341 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5401 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5461 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5521 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5581 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5641 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5701 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5761 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5821 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5881 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5941 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6001 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6061 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6121 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6181 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6241 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6301 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6361 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6421 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6481 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6541 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6601 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6661 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6721 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6781 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6841 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6901 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6961 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7021 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7081 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7141 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7201 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7261 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7321 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7381 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7441 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     7501 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     7561 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     7621 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     7681 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     7741 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     7801 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     7861 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcgc aaggaatggt
     7921 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
     7981 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
     8041 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
     8101 gcgtagaggc gattagtcca atttgttaaa gacaggatat cagtggtcca ggctctagtt
     8161 ttgactcaac aatatcacca gctgaagcct atagagtacg agccatagat aaaataaaag
     8221 attttattta gtctccagaa aaagggggg
//
