CD22
CD22 was amplified from plasmid pRKm22 using the following oligomers: 22COD 5'GTGAGTGAGAATTCATGCATCTCCTCGGCCCCTG-3' and 33NON 5'-TCACTCACTCGCGGCCGCTTCGCCTGCCGATGGTCTC-3'. pRKm22 is a plasmid encoding full-length human CD22 obtained by cloning from a Daudi cDNA Quick clone library (Clontech). The oligomers introduced EcoRI and NotI restriction sites. After digestion with NotI and EcoRI, the PCR product was purified and cloned into vector pCDNA1.1-Fc between NotI and EcoRI sites creating plasmid pCDNA1.1–22-Fc. The 293T cells were transfected with plasmid pCDNA1.1– 22-Fc by standard CaPO4 precipitation
Human Siglec-2 (176-687), His Tag (SI2-H5228) is expressed from human 293 cells (HEK293). It contains AA Trp 176 - Arg 687 (Accession # P20273-1). Predicted N-terminus: Trp 176
Siglec-2(Trp 176 - Arg 687) Poly-his P20273-1
This protein carries a polyhistidine tag at the C-terminus.
The protein has a calculated MW of 57.8 kDa. The protein migrates as 90-100 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.
http://www.uniprot.org/uniprot/P20273 20 – 687 Extracellular 668aa
DSSKWVFEHPETLYAWEGACVWIPCTYRALDGDLESFILFHNPEYNKNTSKFDGTRLYES TKDGKVPSEQKRVQFLGDKNKNCTLSIHPVHLNDSGQLGLRMESKTEKWMERIHLNVSER PFPPHIQLPPEIQESQEVTLTCLLNFSCYGYPIQLQWLLEGVPMRQAAVTSTSLTIKSVF TRSELKFSPQWSHHGKIVTCQLQDADGKFLSNDTVQLNVKHTPKLEIKVTPSDAIVREGD SVTMTCEVSSSNPEYTTVSWLKDGTSLKKQNTFTLNLREVTKDQSGKYCCQVSNDVGPGR SEEVFLQVQYAPEPSTVQILHSPAVEGSQVEFLCMSLANPLPTNYTWYHNGKEMQGRTEE KVHIPKILPWHAGTYSCVAENILGTGQRGPGAELDVQYPPKKVTTVIQNPMPIREGDTVT LSCNYNSSNPSVTRYEWKPHGAWEEPSLGVLKIQNVGWDNTTIACAACNSWCSWASPVAL NVQYAPRDVRVRKIKPLSEIHSGNSVSLQCDFSSSHPKEVQFFWEKNGRLLGKESQLNFD SISPEDAGSYSCWVNNSIGQTASKAWTLEVLYAPRRLRVSMSPGDQVMEGKSATLTCESD ANPPVSHYTWFDWNNQSLPYHSQKLRLEPVKVQHSGAYWCQGTNSVGKGRSPLSTLTVYY SPETIGRR
https://www.ncbi.nlm.nih.gov/nuccore/297374827
ATGCATCTCCTCGGCCCCTGGCTCCTGCTCCTGGTTCTAGAATACTTGGCTTTCTCTGACTCAAGTAAATGGGTTTTTGAGCACCCTGAAACCCTCTACGCCTGGGAGGGGGCCTGCGTCTGGATCCCCTGCACCTACAGAGCCCTAGATGGTGACCTGGAAAGCTTCATCCTGTTCCACAATCCTGAGTATAACAAGAACACCTCGAAGTTTGATGGGACAAGACTCTATGAAAGCACAAAGGATGGGAAGGTTCCTTCTGAGCAGAAAAGGGTGCAATTCCTGGGAGACAAGAATAAGAACTGCACACTGAGTATCCACCCGGTGCACCTCAATGACAGTGGTCAGCTGGGGCTGAGGATGGAGTCCAAGACTGAGAAATGGATGGAACGAATACACCTCAATGTCTCTGAAAGGCCTTTTCCACCTCATATCCAGCTCCCTCCAGAAATTCAAGAGTCCCAGGAAGTCACTCTGACCTGCTTGCTGAATTTCTCCTGCTATGGGTATCCGATCCAATTGCAGTGGCTCCTAGAGGGGGTTCCAATGAGGCAGGCTGCTGTCACCTCGACCTCCTTGACCATCAAGTCTGTCTTCACCCGGAGCGAGCTCAAGTTCTCCCCACAGTGGAGTCACCATGGGAAGATTGTGACCTGCCAGCTTCAGGATGCAGATGGGAAGTTCCTCTCCAATGACACGGTGCAGCTGAACGTGAAGCACACCCCGAAGTTGGAGATCAAGGTCACTCCCAGTGATGCCATAGTGAGGGAGGGGGACTCTGTGACCATGACCTGCGAGGTCAGCAGCAGCAACCCGGAGTACACGACGGTATCCTGGCTCAAGGATGGGACCTCGCTGAAGAAGCAGAATACATTCACGCTAAACCTGCGCGAAGTGACCAAGGACCAGAGTGGGAAGTACTGCTGTCAGGTCTCCAATGACGTGGGCCCGGGAAGGTCGGAAGAAGTGTTCCTGCAAGTGCAGTATGCCCCGGAACCTTCCACGGTTCAGATCCTCCACTCACCGGCTGTGGAGGGAAGTCAAGTCGAGTTTCTTTGCATGTCACTGGCCAATCCTCTTCCAACAAATTACACGTGGTACCACAATGGGAAAGAAATGCAGGGAAGGACAGAGGAGAAAGTCCACATCCCAAAGATCCTCCCCTGGCACGCTGGGACTTATTCCTGTGTGGCAGAAAACATTCTTGGTACTGGACAGAGGGGCCCGGGAGCTGAGCTGGATGTCCAGTATCCTCCCAAGAAGGTGACCACAGTGATTCAAAACCCCATGCCGATTCGAGAAGGAGACACAGTGACCCTTTCCTGTAACTACAATTCCAGTAACCCCAGTGTTACCCGGTATGAATGGAAACCCCATGGCGCCTGGGAGGAGCCATCGCTTGGGGTGCTGAAGATCCAAAACGTTGGCTGGGACAACACAACCATCGCCTGCGCAGCTTGTAATAGTTGGTGCTCGTGGGCCTCCCCTGTCGCCCTGAATGTCCAGTATGCCCCCCGAGACGTGAGGGTCCGGAAAATCAAGCCCCTTTCCGAGATTCACTCTGGAAACTCGGTCAGCCTCCAATGTGACTTCTCAAGCAGCCACCCCAAAGAAGTCCAGTTCTTCTGGGAGAAAAATGGCAGGCTTCTGGGGAAAGAAAGCCAGCTGAATTTTGACTCCATCTCCCCAGAAGATGCTGGGAGTTACAGCTGCTGGGTGAACAACTCCATAGGACAGACAGCGTCCAAGGCCTGGACACTTGAAGTGCTGTATGCACCCAGGAGGCTGCGTGTGTCCATGAGCCCGGGGGACCAAGTGATGGAGGGGAAGAGTGCAACCCTGACCTGTGAGAGCGACGCCAACCCTCCCGTCTCCCACTACACCTGGTTTGACTGGAATAACCAAAGCCTCCCCTACCACAGCCAGAAGCTGAGATTGGAGCCGGTGAAGGTCCAGCACTCGGGTGCCTACTGGTGCCAGGGGACCAACAGTGTGGGCAAGGGCCGTTCGCCTCTCAGCACCCTCACCGTCTACTATAGCCCGGAGACCATCGGCAGGCGA
GTGGCTGTGGGACTCGGGTCCTGCCTCGCCATCCTCATCCTGGCAATCTGTGGGCTCAAGCTCCAGCGACGTTGGAAGAGGACACAGAGCCAGCAGGGGCTTCAGGAGAATTCCAGCGGCCAGAGCTTCTTTGTGAGGAATAAAAAGAGATGCAGAGTCCTCAGAGATGCAGAGACCTCCCCCGGACTGCGATGA
atgcatctcctcggcccctggctcctgctcctggttctagaatacttggctttctctgac M H L L G P W L L L L V L E Y L A F S D tcaagtaaatgggtttttgagcaccctgaaaccctctacgcctgggagggggcctgcgtc S S K W V F E H P E T L Y A W E G A C V tggatcccctgcacctacagagccctagatggtgacctggaaagcttcatcctgttccac W I P C T Y R A L D G D L E S F I L F H aatcctgagtataacaagaacacctcgaagtttgatgggacaagactctatgaaagcaca N P E Y N K N T S K F D G T R L Y E S T aaggatgggaaggttccttctgagcagaaaagggtgcaattcctgggagacaagaataag K D G K V P S E Q K R V Q F L G D K N K aactgcacactgagtatccacccggtgcacctcaatgacagtggtcagctggggctgagg N C T L S I H P V H L N D S G Q L G L R atggagtccaagactgagaaatggatggaacgaatacacctcaatgtctctgaaaggcct M E S K T E K W M E R I H L N V S E R P tttccacctcatatccagctccctccagaaattcaagagtcccaggaagtcactctgacc F P P H I Q L P P E I Q E S Q E V T L T tgcttgctgaatttctcctgctatgggtatccgatccaattgcagtggctcctagagggg C L L N F S C Y G Y P I Q L Q W L L E G gttccaatgaggcaggctgctgtcacctcgacctccttgaccatcaagtctgtcttcacc V P M R Q A A V T S T S L T I K S V F T cggagcgagctcaagttctccccacagtggagtcaccatgggaagattgtgacctgccag R S E L K F S P Q W S H H G K I V T C Q cttcaggatgcagatgggaagttcctctccaatgacacggtgcagctgaacgtgaagcac L Q D A D G K F L S N D T V Q L N V K H accccgaagttggagatcaaggtcactcccagtgatgccatagtgagggagggggactct T P K L E I K V T P S D A I V R E G D S gtgaccatgacctgcgaggtcagcagcagcaacccggagtacacgacggtatcctggctc V T M T C E V S S S N P E Y T T V S W L aaggatgggacctcgctgaagaagcagaatacattcacgctaaacctgcgcgaagtgacc K D G T S L K K Q N T F T L N L R E V T aaggaccagagtgggaagtactgctgtcaggtctccaatgacgtgggcccgggaaggtcg K D Q S G K Y C C Q V S N D V G P G R S gaagaagtgttcctgcaagtgcagtatgccccggaaccttccacggttcagatcctccac E E V F L Q V Q Y A P E P S T V Q I L H tcaccggctgtggagggaagtcaagtcgagtttctttgcatgtcactggccaatcctctt S P A V E G S Q V E F L C M S L A N P L ccaacaaattacacgtggtaccacaatgggaaagaaatgcagggaaggacagaggagaaa P T N Y T W Y H N G K E M Q G R T E E K gtccacatcccaaagatcctcccctggcacgctgggacttattcctgtgtggcagaaaac V H I P K I L P W H A G T Y S C V A E N attcttggtactggacagaggggcccgggagctgagctggatgtccagtatcctcccaag I L G T G Q R G P G A E L D V Q Y P P K aaggtgaccacagtgattcaaaaccccatgccgattcgagaaggagacacagtgaccctt K V T T V I Q N P M P I R E G D T V T L tcctgtaactacaattccagtaaccccagtgttacccggtatgaatggaaaccccatggc S C N Y N S S N P S V T R Y E W K P H G gcctgggaggagccatcgcttggggtgctgaagatccaaaacgttggctgggacaacaca A W E E P S L G V L K I Q N V G W D N T accatcgcctgcgcagcttgtaatagttggtgctcgtgggcctcccctgtcgccctgaat T I A C A A C N S W C S W A S P V A L N gtccagtatgccccccgagacgtgagggtccggaaaatcaagcccctttccgagattcac V Q Y A P R D V R V R K I K P L S E I H tctggaaactcggtcagcctccaatgtgacttctcaagcagccaccccaaagaagtccag S G N S V S L Q C D F S S S H P K E V Q ttcttctgggagaaaaatggcaggcttctggggaaagaaagccagctgaattttgactcc F F W E K N G R L L G K E S Q L N F D S atctccccagaagatgctgggagttacagctgctgggtgaacaactccataggacagaca I S P E D A G S Y S C W V N N S I G Q T gcgtccaaggcctggacacttgaagtgctgtatgcacccaggaggctgcgtgtgtccatg A S K A W T L E V L Y A P R R L R V S M agcccgggggaccaagtgatggaggggaagagtgcaaccctgacctgtgagagcgacgcc S P G D Q V M E G K S A T L T C E S D A aaccctcccgtctcccactacacctggtttgactggaataaccaaagcctcccctaccac N P P V S H Y T W F D W N N Q S L P Y H agccagaagctgagattggagccggtgaaggtccagcactcgggtgcctactggtgccag S Q K L R L E P V K V Q H S G A Y W C Q gggaccaacagtgtgggcaagggccgttcgcctctcagcaccctcaccgtctactatagc G T N S V G K G R S P L S T L T V Y Y S ccggagaccatcggcaggcga P E T I G R R gtggctgtgggactcgggtcctgcctcgccatcctcatc V A V G L G S C L A I L I ctggcaatctgtgggctcaagctccagcgacgttggaagaggacacagagccagcagggg L A I C G L K L Q R R W K R T Q S Q Q G cttcaggagaattccagcggccagagcttctttgtgaggaataaaaagagatgcagagtc L Q E N S S G Q S F F V R N K K R C R V ctcagagatgcagagacctcccccggactgcgatga L R D A E T S P G L R -
688 – 706 Transmembrane Helical 19aa 707 – 847 Cytoplasmic 141aa