LOCUS       sequence-410663-        2702 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses Spycatcher003 on the cell surface using mouse CD80 (short)
            hinge, transmembrane, and cytoplasmic domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2702)
  AUTHORS   Patel A, Andre V, Eguiguren SB, Barton MI, Burton J, Denham EM, 
            Pettmann J, Morch AM, Kutuzov MA, Siller-Farfan JA, Dustin ML, van 
            der Merwe PA, Dushek O
  TITLE     Using CombiCells, a platform for titration and combinatorial display
            of cell surface ligands, to study T-cell antigen sensitivity 
            modulation by accessory receptors.
  JOURNAL   EMBO J. 2024 Jan;43(1):132-150. doi: 10.1038/s44318-023-00012-1. 
            Epub 2023 Dec 18.
  PUBMED    38177315
REFERENCE   2  (bases 1 to 2702)
  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/s44318-023-00012-1"; journalName: "EMBO J"; date: 
            "2024-01"; volume: "43"; issue: "1"; pages: "132-150"
COMMENT     Sequence Label: pHR-SIN-BX SpyCatcher003-(short)mCD80
FEATURES             Location/Qualifiers
     source          1..2702
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        125..531
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      551..560
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     557..616
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             620..958
                     /codon_start=1
                     /product="improved SpyCatcher variant that reacts with 
                     SpyTag003 at a rate approaching the diffusion limit (Keeble
                     et al., 2019)"
                     /label=SpyCatcher003
                     /translation="VTTLSGLSGEQGPSGDMTTEEDSATHIKFSKRDEDGRELAGATME
                     LRDSSGKTISTWISDGHVKDFYLYPGKYTFVETAAPDGYEVATPIEFTVNEDGQVTVDG
                     EATEGDAHT"
     misc_feature    1243..1831
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1296..1316)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1714..1725)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1834..1850)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1835..1851)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             1945..2178
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(2203..2221)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2204..2221)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2511..2530)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 agtttggatc gataagcttg atatcgaatt cctgcagccc cgataaaata aaagatttta
       61 tttagtctcc agaaaaaggg gggaatgaaa gacccccacc tgtaggtttg gcaagctagc
      121 tgcagtaacg ccatttgcaa ggcatggaaa aataccaaac caagaataga gaagttcaga
      181 tcaagggcgg gtacatgaaa atagctaacg ttgggccaaa caggatatct gcggtgagca
      241 gtttcggccc cggcccgggg ccaagaacag atggtcaccg cagtttcggc cccggcccga
      301 ggccaagaac agatggtccc cagatatggc ccaaccctca gcagtttctt aagacccatc
      361 agatgtttcc aggctccccc aaggacctga aatgaccctg cgccttattt gaattaacca
      421 atcagcctgc ttctcgcttc tgttcgcgcg cttctgcttc ccgagctcta taaaagagct
      481 cacaacccct cactcggcgc gccagtcctc cgacagactg agtcgcccgg gggggatccc
      541 ggtactcgag gccaccatgg agacagacac actcctgcta tgggtactgc tgctctgggt
      601 tccaggttcc actggtgacg tgaccacact gtccggactg tctggagagc agggaccatc
      661 cggcgacatg accacagagg aggattctgc cacacacatc aagttcagca agagggacga
      721 ggacggaaga gagctggcag gagcaaccat ggagctgagg gatagctccg gcaagaccat
      781 cagcacatgg atctccgacg gccacgtgaa ggatttctac ctgtatcccg gcaagtacac
      841 ctttgtggag acagcagcac cagacggata tgaggtggca acccctatcg agtttacagt
      901 gaacgaggac ggacaggtga ccgtggatgg agaggcaaca gagggcgatg cacacaccgg
      961 atctagcgga tctggtggaa gccctcctga tagcaagaac acacttgtgc tctttggggc
     1021 aggattcggc gcagtaataa cagtcgtcgt catcgttgtc atcatcaaat gcttctgtaa
     1081 gcacagaagc tgtttcagaa gaaatgaggc aagcagagaa acaaacaaca gccttacctt
     1141 cgggcctgaa gaagcattag ctgaacagac cgtcttcctt taggcggccg cgactctaga
     1201 gtcgacctgc aggcatgcaa gcttgatatc aagcttatcg ataatcaacc tctggattac
     1261 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     1321 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     1381 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     1441 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     1501 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     1561 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     1621 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     1681 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     1741 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     1801 agtcggatct ccctttgggc cgcctccccg catcgatacc gtcgacctcg atcgagggaa
     1861 ttaattcgag ctcggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     1921 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     1981 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     2041 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     2101 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     2161 tgtggaaaat ctctagcagc atctagaatt aattccgtgt attctatagt gtcacctaaa
     2221 tcgtatgtgt atgatacata aggttatgta ttaattgtag ccgcgttcta acgacaatat
     2281 gtacaagcct aattgtgtag catctggctt actgaagcag accctatcat ctctctcgta
     2341 aactgccgtc agagtcggtt tggttggacg aaccttctga gtttctggta acgccgtccc
     2401 gcacccggaa atggtcagcg aaccaatcag cagggtcatc gctagccaga tcctctacgc
     2461 cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc
     2521 cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg
     2581 cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc
     2641 accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat
     2701 gc
//
