
                                SEQUENCE LISTING

<110> The General Hospital Corporation

<120> Anti-human Chemokine (C-C motif)
  Receptor 4 Immunotoxins

<130> 29539-0132WO1

<140> PCT/US2015/035550   
<141> 2015-06-12  

<150> US 62/049,096       
<151> 2014-09-11  

<150> US 62/011,235       
<151> 2014-06-12  

<160> 16

<170> FastSEQ for Windows Version 4.0

<210> 1
<211> 360
<212> PRT
<213> Homo sapiens

<400> 1
Met Asn Pro Thr Asp Ile Ala Asp Thr Thr Leu Asp Glu Ser Ile Tyr
 1               5                  10                  15      
Ser Asn Tyr Tyr Leu Tyr Glu Ser Ile Pro Lys Pro Cys Thr Lys Glu
            20                  25                  30          
Gly Ile Lys Ala Phe Gly Glu Leu Phe Leu Pro Pro Leu Tyr Ser Leu
        35                  40                  45              
Val Phe Val Phe Gly Leu Leu Gly Asn Ser Val Val Val Leu Val Leu
    50                  55                  60                  
Phe Lys Tyr Lys Arg Leu Arg Ser Met Thr Asp Val Tyr Leu Leu Asn
65                  70                  75                  80  
Leu Ala Ile Ser Asp Leu Leu Phe Val Phe Ser Leu Pro Phe Trp Gly
                85                  90                  95      
Tyr Tyr Ala Ala Asp Gln Trp Val Phe Gly Leu Gly Leu Cys Lys Met
            100                 105                 110         
Ile Ser Trp Met Tyr Leu Val Gly Phe Tyr Ser Gly Ile Phe Phe Val
        115                 120                 125             
Met Leu Met Ser Ile Asp Arg Tyr Leu Ala Ile Val His Ala Val Phe
    130                 135                 140                 
Ser Leu Arg Ala Arg Thr Leu Thr Tyr Gly Val Ile Thr Ser Leu Ala
145                 150                 155                 160 
Thr Trp Ser Val Ala Val Phe Ala Ser Leu Pro Gly Phe Leu Phe Ser
                165                 170                 175     
Thr Cys Tyr Thr Glu Arg Asn His Thr Tyr Cys Lys Thr Lys Tyr Ser
            180                 185                 190         
Leu Asn Ser Thr Thr Trp Lys Val Leu Ser Ser Leu Glu Ile Asn Ile
        195                 200                 205             
Leu Gly Leu Val Ile Pro Leu Gly Ile Met Leu Phe Cys Tyr Ser Met
    210                 215                 220                 
Ile Ile Arg Thr Leu Gln His Cys Lys Asn Glu Lys Lys Asn Lys Ala
225                 230                 235                 240 
Val Lys Met Ile Phe Ala Val Val Val Leu Phe Leu Gly Phe Trp Thr
                245                 250                 255     
Pro Tyr Asn Ile Val Leu Phe Leu Glu Thr Leu Val Glu Leu Glu Val
            260                 265                 270         
Leu Gln Asp Cys Thr Phe Glu Arg Tyr Leu Asp Tyr Ala Ile Gln Ala
        275                 280                 285             
Thr Glu Thr Leu Ala Phe Val His Cys Cys Leu Asn Pro Ile Ile Tyr
    290                 295                 300                 
Phe Phe Leu Gly Glu Lys Phe Arg Lys Tyr Ile Leu Gln Leu Phe Lys
305                 310                 315                 320 
Thr Cys Arg Gly Leu Phe Val Leu Cys Gln Tyr Cys Gly Leu Leu Gln
                325                 330                 335     
Ile Tyr Ser Ala Asp Thr Pro Ser Ser Ser Tyr Thr Gln Ser Thr Met
            340                 345                 350         
Asp His Asp Leu His Asp Ala Leu
        355                 360 


<210> 2
<211> 8
<212> PRT
<213> Artificial Sequence

<220> 
<223> FLAG peptide

<400> 2
Asp Tyr Lys Asp Asp Asp Asp Lys
 1               5              


<210> 3
<211> 6
<212> PRT
<213> Artificial Sequence

<220> 
<223> Hemagglutinin peptide

<400> 3
Tyr Pro Tyr Asp Val Pro
 1               5      


<210> 4
<211> 10
<212> PRT
<213> Artificial Sequence

<220> 
<223> proto-oncoprotein myc tag

<400> 4
Ile Leu Lys Lys Ala Thr Ala Tyr Ile Leu
 1               5                  10  


<210> 5
<211> 10
<212> PRT
<213> Artificial Sequence

<220> 
<223> proto-oncoprotein myc tag

<400> 5
Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu
 1               5                  10  


<210> 6
<211> 735
<212> DNA
<213> Artificial Sequence

<220> 
<223> codon-optimized cDNA encoding an anti-human CCR4
      scFv

<400> 6
gacattgagt tgactcaatc tccatcttcc ttggctgttt ctgctggtga gaaggttact 60
atgtcttgta agtcttccca atctattttg tactcttcca accaaaagaa ctacttggct 120
tggtaccaac aaaagccagg tcaatctcca aagttgttga tttactgggc ttctactaga 180
gagtctggtg ttccagacag attcactggt tctggttctg gtactgactt cactttgact 240
atttcttccg ttcaagctga ggacttggct gtttactact gtcaccaata cttgtcttcc 300
tacactttcg gtggtggtac taagttggag attaagggtg gtggtggttc tggtggtggt 360
ggatctggtg gtggtggttc tcaagttcaa ttgcaacaat ctggtccaga gttggttaga 420
ccaggtgctt ctgttagaat ttcttgtaag gcttctggtt acactttcgc ttcttactac 480
attcaatgga tgaagcaaag accaggtcaa ggtttggagt ggattggttg gattaaccca 540
ggtaacgtta acactaagta caacgagaag ttcaagggta aggctacttt gactgctgac 600
aagtcttcca ctaccgctta catgcaattg tcttccttga cttctgagga ctctgctgtt 660
tacttctgtg ctagatccac ttactacaga ccattggact actggggtca aggtactacc 720
gttactgttt cttcc                                                  735

<210> 7
<211> 245
<212> PRT
<213> Artificial Sequence

<220> 
<223> anti-human CCR4 scFv

<400> 7
Asp Ile Glu Leu Thr Gln Ser Pro Ser Ser Leu Ala Val Ser Ala Gly
 1               5                  10                  15      
Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Ile Leu Tyr Ser
            20                  25                  30          
Ser Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln
        35                  40                  45              
Ser Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val
    50                  55                  60                  
Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
65                  70                  75                  80  
Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys His Gln
                85                  90                  95      
Tyr Leu Ser Ser Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
            100                 105                 110         
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln
        115                 120                 125             
Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Arg Pro Gly Ala Ser
    130                 135                 140                 
Val Arg Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ala Ser Tyr Tyr
145                 150                 155                 160 
Ile Gln Trp Met Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile Gly
                165                 170                 175     
Trp Ile Asn Pro Gly Asn Val Asn Thr Lys Tyr Asn Glu Lys Phe Lys
            180                 185                 190         
Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Thr Thr Ala Tyr Met
        195                 200                 205             
Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala
    210                 215                 220                 
Arg Ser Thr Tyr Tyr Arg Pro Leu Asp Tyr Trp Gly Gln Gly Thr Thr
225                 230                 235                 240 
Val Thr Val Ser Ser
                245 


<210> 8
<211> 705
<212> DNA
<213> Artificial Sequence

<220> 
<223> codon-optimized cDNA encoding an anti-human CCR4
      scFv

<400> 8
gacattgagt tgactcaatc tccatcttcc ttggctgttt ctgctggtga gaaggttact 60
atgtcttgta agtcttccca atctattttg tactcttcca accaaaagaa ctacttggct 120
tggtaccaac aaaagccagg tcaatctcca aagttgttga tttactgggc ttctactaga 180
gagtctggtg ttccagacag attcactggt tctggttctg gtactgactt cactttgact 240
atttcttccg ttcaagctga ggacttggct gtttactact gtcaccaata cttgtcttcc 300
tacactttcg gtggtggtac taagttggag attaagggtg gtggtggatc tcaagttcaa 360
ttgcaacaat ctggtccaga gttggttaga ccaggtgctt ctgttagaat ttcttgtaag 420
gcttctggtt acactttcgc ttcttactac attcaatgga tgaagcaaag accaggtcaa 480
ggtttggagt ggattggttg gattaaccca ggtaacgtta acactaagta caacgagaag 540
ttcaagggta aggctacttt gactgctgac aagtcttcca ctaccgctta catgcaattg 600
tcttccttga cttctgagga ctctgctgtt tacttctgtg ctagatccac ttactacaga 660
ccattggact actggggtca aggtactacc gttactgttt cttcc                 705

<210> 9
<211> 235
<212> PRT
<213> Artificial Sequence

<220> 
<223> anti-human CCR4 scFv

<400> 9
Asp Ile Glu Leu Thr Gln Ser Pro Ser Ser Leu Ala Val Ser Ala Gly
 1               5                  10                  15      
Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Ile Leu Tyr Ser
            20                  25                  30          
Ser Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln
        35                  40                  45              
Ser Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val
    50                  55                  60                  
Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
65                  70                  75                  80  
Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys His Gln
                85                  90                  95      
Tyr Leu Ser Ser Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
            100                 105                 110         
Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Gln Ser Gly Pro Glu Leu
        115                 120                 125             
Val Arg Pro Gly Ala Ser Val Arg Ile Ser Cys Lys Ala Ser Gly Tyr
    130                 135                 140                 
Thr Phe Ala Ser Tyr Tyr Ile Gln Trp Met Lys Gln Arg Pro Gly Gln
145                 150                 155                 160 
Gly Leu Glu Trp Ile Gly Trp Ile Asn Pro Gly Asn Val Asn Thr Lys
                165                 170                 175     
Tyr Asn Glu Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser
            180                 185                 190         
Ser Thr Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser
        195                 200                 205             
Ala Val Tyr Phe Cys Ala Arg Ser Thr Tyr Tyr Arg Pro Leu Asp Tyr
    210                 215                 220                 
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser
225                 230                 235 


<210> 10
<211> 54
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetically generated primer

<400> 10
ccgctcgagc catggggtgg tggtggttct gacattgagt tgactcaatc tcca       54

<210> 11
<211> 66
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetically generated primer

<400> 11
ccggaattcc gccgcggatc caccaccacc agaaccacca ccaccggaag aaacagtaac 60
ggtagt                                                            66

<210> 12
<211> 54
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetically generated primer

<400> 12
ccgctcgagg gatccggtgg tggtggttct gacattgagt tgactcaatc tcca       54

<210> 13
<211> 51
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetically generated primer

<400> 13
ccggaattct tagtggtggt ggtggtggtg ggaagaaaca gtaacggtag t          51

<210> 14
<211> 41
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetically generated primer

<400> 14
cgcggatcca ccaccaccct taatctccaa cttagtacca c                     41

<210> 15
<211> 32
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetically generated primer

<400> 15
ggaagatctc aagttcaatt gcaacaatct gg                               32

<210> 16
<211> 5
<212> PRT
<213> Artificial Sequence

<220> 
<223> linker

<400> 16
Gly Gly Gly Gly Ser
 1               5  

