LOCUS       sequence-410660-        2603 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses Spycatcher003 on the cell surface using human CD52 hinge.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2603)
  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 2603)
  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-hCD52
FEATURES             Location/Qualifiers
     source          1..2603
                     /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    1144..1732
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1197..1217)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1615..1626)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1735..1751)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1736..1752)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             1846..2079
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(2104..2122)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2105..2122)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2412..2431)
                     /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 gcaccagcca aaccagcagc ccctcagcat ccagcaacat
     1021 aagcggaggc attttccttt tcttcgtggc caatgccata atccacctct tctgcttcag
     1081 ttaggcggcc gcgactctag agtcgacctg caggcatgca agcttgatat caagcttatc
     1141 gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     1201 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     1261 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     1321 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     1381 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     1441 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     1501 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     1561 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     1621 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     1681 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac
     1741 cgtcgacctc gatcgaggga attaattcga gctcggtacc tttaagacca atgacttaca
     1801 aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc
     1861 actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga
     1921 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     1981 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     2041 ccctcagacc cttttagtca gtgtggaaaa tctctagcag catctagaat taattccgtg
     2101 tattctatag tgtcacctaa atcgtatgtg tatgatacat aaggttatgt attaattgta
     2161 gccgcgttct aacgacaata tgtacaagcc taattgtgta gcatctggct tactgaagca
     2221 gaccctatca tctctctcgt aaactgccgt cagagtcggt ttggttggac gaaccttctg
     2281 agtttctggt aacgccgtcc cgcacccgga aatggtcagc gaaccaatca gcagggtcat
     2341 cgctagccag atcctctacg ccggacgcat cgtggccggc atcaccggcg ccacaggtgc
     2401 ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc gccacttcgg
     2461 gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt
     2521 gggcgccatc tccttgcatg caccattcct tgcggcggcg gtgctcaacg gcctcaacct
     2581 actactgggc tgcttcctaa tgc
//
