Editing 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 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 https://www.ncbi.nlm.nih.gov/nuccore/297374827]

ATGCATCTCCTCGGCCCCTGGCTCCTGCTCCTGGTTCTAGAATACTTGGCTTTCTCTGACTCAAGTAAATGGGTTTTTGAGCACCCTGAAACCCTCTACGCCTGGGAGGGGGCCTGCGTCTGGATCCCCTGCACCTACAGAGCCCTAGATGGTGACCTGGAAAGCTTCATCCTGTTCCACAATCCTGAGTATAACAAGAACACCTCGAAGTTTGATGGGACAAGACTCTATGAAAGCACAAAGGATGGGAAGGTTCCTTCTGAGCAGAAAAGGGTGCAATTCCTGGGAGACAAGAATAAGAACTGCACACTGAGTATCCACCCGGTGCACCTCAATGACAGTGGTCAGCTGGGGCTGAGGATGGAGTCCAAGACTGAGAAATGGATGGAACGAATACACCTCAATGTCTCTGAAAGGCCTTTTCCACCTCATATCCAGCTCCCTCCAGAAATTCAAGAGTCCCAGGAAGTCACTCTGACCTGCTTGCTGAATTTCTCCTGCTATGGGTATCCGATCCAATTGCAGTGGCTCCTAGAGGGGGTTCCAATGAGGCAGGCTGCTGTCACCTCGACCTCCTTGACCATCAAGTCTGTCTTCACCCGGAGCGAGCTCAAGTTCTCCCCACAGTGGAGTCACCATGGGAAGATTGTGACCTGCCAGCTTCAGGATGCAGATGGGAAGTTCCTCTCCAATGACACGGTGCAGCTGAACGTGAAGCACACCCCGAAGTTGGAGATCAAGGTCACTCCCAGTGATGCCATAGTGAGGGAGGGGGACTCTGTGACCATGACCTGCGAGGTCAGCAGCAGCAACCCGGAGTACACGACGGTATCCTGGCTCAAGGATGGGACCTCGCTGAAGAAGCAGAATACATTCACGCTAAACCTGCGCGAAGTGACCAAGGACCAGAGTGGGAAGTACTGCTGTCAGGTCTCCAATGACGTGGGCCCGGGAAGGTCGGAAGAAGTGTTCCTGCAAGTGCAGTATGCCCCGGAACCTTCCACGGTTCAGATCCTCCACTCACCGGCTGTGGAGGGAAGTCAAGTCGAGTTTCTTTGCATGTCACTGGCCAATCCTCTTCCAACAAATTACACGTGGTACCACAATGGGAAAGAAATGCAGGGAAGGACAGAGGAGAAAGTCCACATCCCAAAGATCCTCCCCTGGCACGCTGGGACTTATTCCTGTGTGGCAGAAAACATTCTTGGTACTGGACAGAGGGGCCCGGGAGCTGAGCTGGATGTCCAGTATCCTCCCAAGAAGGTGACCACAGTGATTCAAAACCCCATGCCGATTCGAGAAGGAGACACAGTGACCCTTTCCTGTAACTACAATTCCAGTAACCCCAGTGTTACCCGGTATGAATGGAAACCCCATGGCGCCTGGGAGGAGCCATCGCTTGGGGTGCTGAAGATCCAAAACGTTGGCTGGGACAACACAACCATCGCCTGCGCAGCTTGTAATAGTTGGTGCTCGTGGGCCTCCCCTGTCGCCCTGAATGTCCAGTATGCCCCCCGAGACGTGAGGGTCCGGAAAATCAAGCCCCTTTCCGAGATTCACTCTGGAAACTCGGTCAGCCTCCAATGTGACTTCTCAAGCAGCCACCCCAAAGAAGTCCAGTTCTTCTGGGAGAAAAATGGCAGGCTTCTGGGGAAAGAAAGCCAGCTGAATTTTGACTCCATCTCCCCAGAAGATGCTGGGAGTTACAGCTGCTGGGTGAACAACTCCATAGGACAGACAGCGTCCAAGGCCTGGACACTTGAAGTGCTGTATGCACCCAGGAGGCTGCGTGTGTCCATGAGCCCGGGGGACCAAGTGATGGAGGGGAAGAGTGCAACCCTGACCTGTGAGAGCGACGCCAACCCTCCCGTCTCCCACTACACCTGGTTTGACTGGAATAACCAAAGCCTCCCCTACCACAGCCAGAAGCTGAGATTGGAGCCGGTGAAGGTCCAGCACTCGGGTGCCTACTGGTGCCAGGGGACCAACAGTGTGGGCAAGGGCCGTTCGCCTCTCAGCACCCTCACCGTCTACTATAGCCCGGAGACCATCGGCAGGCGA



GTGGCTGTGGGACTCGGGTCCTGCCTCGCCATCCTCATCCTGGCAATCTGTGGGCTCAAGCTCCAGCGACGTTGGAAGAGGACACAGAGCCAGCAGGGGCTTCAGGAGAATTCCAGCGGCCAGAGCTTCTTTGTGAGGAATAAAAAGAGATGCAGAGTCCTCAGAGATGCAGAGACCTCCCCCGGACTGCGATGA
<pre>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  -  
</pre>









688 – 706 Transmembrane Helical 19aa 707 – 847 Cytoplasmic 141aa
[[Category:Antibodies]]