               SEQUENCE LISTING

<110> Academisch Medisch Centrum

<120> Improved HIV envelope glycoprotein immunogens

<130> 2016-138EP-PR

<140> EP17188050
<141> 2017-08-26

<150> EP17188050.3
<151> 2017-08-26

<160> 18

<170> BiSSAP 1.3.6

<210> 1
<211> 664
<212> PRT
<213> Artificial Sequence


<220> 
<223> HxB2

<400> 1
Met Arg Val Lys Glu Lys Tyr Gln His Leu Trp Arg Trp Gly Trp Arg 
1               5                   10                  15      
Trp Gly Thr Met Leu Leu Gly Met Leu Met Ile Cys Ser Ala Thr Glu 
            20                  25                  30          
Lys Leu Trp Val Thr Val Tyr Tyr Gly Val Pro Val Trp Lys Glu Ala 
        35                  40                  45              
Thr Thr Thr Leu Phe Cys Ala Ser Asp Ala Lys Ala Tyr Asp Thr Glu 
    50                  55                  60                  
Val His Asn Val Trp Ala Thr His Ala Cys Val Pro Thr Asp Pro Asn 
65                  70                  75                  80  
Pro Gln Glu Val Val Leu Val Asn Val Thr Glu Asn Phe Asn Met Trp 
                85                  90                  95      
Lys Asn Asp Met Val Glu Gln Met His Glu Asp Ile Ile Ser Leu Trp 
            100                 105                 110         
Asp Gln Ser Leu Lys Pro Cys Val Lys Leu Thr Pro Leu Cys Val Ser 
        115                 120                 125             
Leu Lys Cys Thr Asp Leu Lys Asn Asp Thr Asn Thr Asn Ser Ser Ser 
    130                 135                 140                 
Gly Arg Met Ile Met Glu Lys Gly Glu Ile Lys Asn Cys Ser Phe Asn 
145                 150                 155                 160 
Ile Ser Thr Ser Ile Arg Gly Lys Val Gln Lys Glu Tyr Ala Phe Phe 
                165                 170                 175     
Tyr Lys Leu Asp Ile Ile Pro Ile Asp Asn Asp Thr Thr Ser Tyr Lys 
            180                 185                 190         
Leu Thr Ser Cys Asn Thr Ser Val Ile Thr Gln Ala Cys Pro Lys Val 
        195                 200                 205             
Ser Phe Glu Pro Ile Pro Ile His Tyr Cys Ala Pro Ala Gly Phe Ala 
    210                 215                 220                 
Ile Leu Lys Cys Asn Asn Lys Thr Phe Asn Gly Thr Gly Pro Cys Thr 
225                 230                 235                 240 
Asn Val Ser Thr Val Gln Cys Thr His Gly Ile Arg Pro Val Val Ser 
                245                 250                 255     
Thr Gln Leu Leu Leu Asn Gly Ser Leu Ala Glu Glu Glu Val Val Ile 
            260                 265                 270         
Arg Ser Val Asn Phe Thr Asp Asn Ala Lys Thr Ile Ile Val Gln Leu 
        275                 280                 285             
Asn Thr Ser Val Glu Ile Asn Cys Thr Arg Pro Asn Asn Asn Thr Arg 
    290                 295                 300                 
Lys Arg Ile Arg Ile Gln Arg Gly Pro Gly Arg Ala Phe Val Thr Ile 
305                 310                 315                 320 
Gly Lys Ile Gly Asn Met Arg Gln Ala His Cys Asn Ile Ser Arg Ala 
                325                 330                 335     
Lys Trp Asn Asn Thr Leu Lys Gln Ile Ala Ser Lys Leu Arg Glu Gln 
            340                 345                 350         
Phe Gly Asn Asn Lys Thr Ile Ile Phe Lys Gln Ser Ser Gly Gly Asp 
        355                 360                 365             
Pro Glu Ile Val Thr His Ser Phe Asn Cys Gly Gly Glu Phe Phe Tyr 
    370                 375                 380                 
Cys Asn Ser Thr Gln Leu Phe Asn Ser Thr Trp Phe Asn Ser Thr Trp 
385                 390                 395                 400 
Ser Thr Glu Gly Ser Asn Asn Thr Glu Gly Ser Asp Thr Ile Thr Leu 
                405                 410                 415     
Pro Cys Arg Ile Lys Gln Ile Ile Asn Met Trp Gln Lys Val Gly Lys 
            420                 425                 430         
Ala Met Tyr Ala Pro Pro Ile Ser Gly Gln Ile Arg Cys Ser Ser Asn 
        435                 440                 445             
Ile Thr Gly Leu Leu Leu Thr Arg Asp Gly Gly Asn Ser Asn Asn Glu 
    450                 455                 460                 
Ser Glu Ile Phe Arg Pro Gly Gly Gly Asp Met Arg Asp Asn Trp Arg 
465                 470                 475                 480 
Ser Glu Leu Tyr Lys Tyr Lys Val Val Lys Ile Glu Pro Leu Gly Val 
                485                 490                 495     
Ala Pro Thr Lys Ala Lys Arg Arg Val Val Gln Arg Glu Lys Arg Ala 
            500                 505                 510         
Val Gly Ile Gly Ala Leu Phe Leu Gly Phe Leu Gly Ala Ala Gly Ser 
        515                 520                 525             
Thr Met Gly Ala Ala Ser Met Thr Leu Thr Val Gln Ala Arg Gln Leu 
    530                 535                 540                 
Leu Ser Gly Ile Val Gln Gln Gln Asn Asn Leu Leu Arg Ala Ile Glu 
545                 550                 555                 560 
Ala Gln Gln His Leu Leu Gln Leu Thr Val Trp Gly Ile Lys Gln Leu 
                565                 570                 575     
Gln Ala Arg Ile Leu Ala Val Glu Arg Tyr Leu Lys Asp Gln Gln Leu 
            580                 585                 590         
Leu Gly Ile Trp Gly Cys Ser Gly Lys Leu Ile Cys Thr Thr Ala Val 
        595                 600                 605             
Pro Trp Asn Ala Ser Trp Ser Asn Lys Ser Leu Glu Gln Ile Trp Asn 
    610                 615                 620                 
His Thr Thr Trp Met Glu Trp Asp Arg Glu Ile Asn Asn Tyr Thr Ser 
625                 630                 635                 640 
Leu Ile His Ser Leu Ile Glu Glu Ser Gln Asn Gln Gln Glu Lys Asn 
                645                 650                 655     
Glu Gln Glu Leu Leu Glu Leu Asp 
            660                 

<210> 2
<211> 634
<212> PRT
<213> Artificial Sequence


<220> 
<223> BG505 SOSIP.v4.1

<400> 2
Ala Glu Asn Leu Trp Val Thr Val Tyr Tyr Gly Val Pro Val Trp Lys 
1               5                   10                  15      
Asp Ala Glu Thr Thr Leu Phe Cys Ala Ser Asp Ala Lys Ala Tyr Glu 
            20                  25                  30          
Thr Lys Lys His Asn Val Trp Ala Thr His Ala Cys Val Pro Thr Asp 
        35                  40                  45              
Pro Asn Pro Gln Glu Ile His Leu Glu Asn Val Thr Glu Glu Phe Asn 
    50                  55                  60                  
Met Trp Lys Asn Asn Met Val Glu Gln Met His Thr Asp Ile Ile Ser 
65                  70                  75                  80  
Leu Trp Asp Gln Ser Leu Lys Pro Cys Val Lys Leu Thr Pro Leu Cys 
                85                  90                  95      
Val Thr Leu Gln Cys Thr Asn Val Thr Asn Asn Ile Thr Asp Asp Met 
            100                 105                 110         
Arg Gly Glu Leu Lys Asn Cys Ser Phe Asn Met Thr Thr Glu Leu Arg 
        115                 120                 125             
Asp Lys Lys Gln Lys Val Tyr Ser Leu Phe Tyr Arg Leu Asp Val Val 
    130                 135                 140                 
Gln Ile Asn Glu Asn Gln Gly Asn Arg Ser Asn Asn Ser Asn Lys Glu 
145                 150                 155                 160 
Tyr Arg Leu Ile Asn Cys Asn Thr Ser Ala Ile Thr Gln Ala Cys Pro 
                165                 170                 175     
Lys Val Ser Phe Glu Pro Ile Pro Ile His Tyr Cys Ala Pro Ala Gly 
            180                 185                 190         
Phe Ala Ile Leu Lys Cys Lys Asp Lys Lys Phe Asn Gly Thr Gly Pro 
        195                 200                 205             
Cys Pro Ser Val Ser Thr Val Gln Cys Thr His Gly Ile Lys Pro Val 
    210                 215                 220                 
Val Ser Thr Gln Leu Leu Leu Asn Gly Ser Leu Ala Glu Glu Glu Val 
225                 230                 235                 240 
Met Ile Arg Ser Glu Asn Ile Thr Asn Asn Ala Lys Asn Ile Leu Val 
                245                 250                 255     
Gln Phe Asn Thr Pro Val Gln Ile Asn Cys Thr Arg Pro Asn Asn Asn 
            260                 265                 270         
Thr Arg Lys Ser Ile Arg Ile Gly Pro Gly Gln Trp Phe Tyr Ala Thr 
        275                 280                 285             
Gly Asp Ile Ile Gly Asp Ile Arg Gln Ala His Cys Asn Val Ser Lys 
    290                 295                 300                 
Ala Thr Trp Asn Glu Thr Leu Gly Lys Val Val Lys Gln Leu Arg Lys 
305                 310                 315                 320 
His Phe Gly Asn Asn Thr Ile Ile Arg Phe Ala Asn Ser Ser Gly Gly 
                325                 330                 335     
Asp Leu Glu Val Thr Thr His Ser Phe Asn Cys Gly Gly Glu Phe Phe 
            340                 345                 350         
Tyr Cys Asn Thr Ser Gly Leu Phe Asn Ser Thr Trp Ile Ser Asn Thr 
        355                 360                 365             
Ser Val Gln Gly Ser Asn Ser Thr Gly Ser Asn Asp Ser Ile Thr Leu 
    370                 375                 380                 
Pro Cys Arg Ile Lys Gln Ile Ile Asn Met Trp Gln Arg Ile Gly Gln 
385                 390                 395                 400 
Ala Met Tyr Ala Pro Pro Ile Gln Gly Val Ile Arg Cys Val Ser Asn 
                405                 410                 415     
Ile Thr Gly Leu Ile Leu Thr Arg Asp Gly Gly Ser Thr Asn Ser Thr 
            420                 425                 430         
Thr Glu Thr Phe Arg Pro Gly Gly Gly Asp Met Arg Asp Asn Trp Arg 
        435                 440                 445             
Ser Glu Leu Tyr Lys Tyr Lys Val Val Lys Ile Glu Pro Leu Gly Val 
    450                 455                 460                 
Ala Pro Thr Arg Cys Lys Arg Arg Val Val Gly Arg Arg Arg Arg Arg 
465                 470                 475                 480 
Arg Ala Val Gly Ile Gly Ala Val Phe Leu Gly Phe Leu Gly Ala Ala 
                485                 490                 495     
Gly Ser Thr Met Gly Ala Ala Ser Met Thr Leu Thr Val Gln Ala Arg 
            500                 505                 510         
Asn Leu Leu Ser Gly Ile Val Gln Gln Gln Ser Asn Leu Leu Arg Ala 
        515                 520                 525             
Pro Glu Ala Gln Gln His Leu Leu Lys Leu Thr Val Trp Gly Ile Lys 
    530                 535                 540                 
Gln Leu Gln Ala Arg Val Leu Ala Val Glu Arg Tyr Leu Arg Asp Gln 
545                 550                 555                 560 
Gln Leu Leu Gly Ile Trp Gly Cys Ser Gly Lys Leu Ile Cys Cys Thr 
                565                 570                 575     
Asn Val Pro Trp Asn Ser Ser Trp Ser Asn Arg Asn Leu Ser Glu Ile 
            580                 585                 590         
Trp Asp Asn Met Thr Trp Leu Gln Trp Asp Lys Glu Ile Ser Asn Tyr 
        595                 600                 605             
Thr Gln Ile Ile Tyr Gly Leu Leu Glu Glu Ser Gln Asn Gln Gln Glu 
    610                 615                 620                 
Lys Asn Glu Gln Asp Leu Leu Ala Leu Asp 
625                 630                 

<210> 3
<211> 627
<212> PRT
<213> Artificial Sequence


<220> 
<223> BG505 SOSIP.v4.1-GT1

<400> 3
Ala Glu Asn Leu Trp Val Thr Val Tyr Tyr Gly Val Pro Val Trp Lys 
1               5                   10                  15      
Asp Ala Glu Thr Thr Leu Phe Cys Ala Ser Asp Ala Lys Ala Tyr Glu 
            20                  25                  30          
Thr Lys Lys His Asn Val Trp Ala Thr His Ala Cys Val Pro Thr Asp 
        35                  40                  45              
Pro Asn Pro Gln Glu Ile His Leu Glu Asn Val Thr Glu Glu Phe Asn 
    50                  55                  60                  
Met Trp Lys Asn Asn Met Val Glu Gln Met His Thr Asp Ile Ile Ser 
65                  70                  75                  80  
Leu Trp Asp Gln Ser Leu Lys Pro Cys Val Lys Leu Thr Pro Leu Cys 
                85                  90                  95      
Val Thr Leu Gln Cys Thr Asn Val Thr Asn Asn Ile Thr Asp Asp Met 
            100                 105                 110         
Arg Gly Glu Leu Lys Asn Cys Ser Phe Asn Met Thr Thr Glu Leu Arg 
        115                 120                 125             
Asp Lys Arg Gln Lys Val His Ala Leu Phe Tyr Lys Leu Asp Ile Val 
    130                 135                 140                 
Pro Ile Asn Glu Asn Gln Asn Thr Ser Tyr Arg Leu Ile Asn Cys Asn 
145                 150                 155                 160 
Thr Ala Ala Ile Thr Gln Ala Cys Pro Lys Val Ser Phe Glu Pro Ile 
                165                 170                 175     
Pro Ile His Tyr Cys Ala Pro Ala Gly Phe Ala Ile Leu Lys Cys Lys 
            180                 185                 190         
Asp Lys Lys Phe Asn Gly Thr Gly Pro Cys Pro Ser Val Ser Thr Val 
        195                 200                 205             
Gln Cys Thr His Gly Ile Lys Pro Val Val Ser Thr Gln Leu Leu Leu 
    210                 215                 220                 
Asn Gly Ser Leu Ala Glu Glu Glu Val Met Ile Arg Ser Lys Asp Ile 
225                 230                 235                 240 
Arg Asn Asn Ala Lys Asn Ile Leu Val Gln Phe Asn Thr Pro Val Gln 
                245                 250                 255     
Ile Asn Cys Thr Arg Pro Asn Asn Asn Thr Arg Lys Ser Ile Arg Ile 
            260                 265                 270         
Gly Pro Gly Gln Trp Phe Tyr Ala Thr Gly Asp Ile Ile Gly Asp Ile 
        275                 280                 285             
Arg Gln Ala His Cys Asn Val Ser Lys Ala Thr Trp Asn Glu Thr Leu 
    290                 295                 300                 
Gly Lys Val Val Lys Gln Leu Arg Lys His Phe Gly Asn Asn Thr Ile 
305                 310                 315                 320 
Ile Arg Phe Ala Asn Ser Ser Gly Gly Asp Leu Glu Val Thr Thr His 
                325                 330                 335     
Ser Phe Asn Cys Gly Gly Glu Phe Phe Tyr Cys Asp Thr Ser Gly Leu 
            340                 345                 350         
Phe Asn Ser Thr Trp Ile Ser Asn Thr Ser Val Gln Gly Ser Asn Ser 
        355                 360                 365             
Thr Gly Ser Asn Asp Ser Ile Thr Leu Pro Cys Arg Ile Lys Gln Ile 
    370                 375                 380                 
Ile Asn Met Trp Gln Arg Ile Gly Gln Ala Met Tyr Ala Pro Pro Ile 
385                 390                 395                 400 
Gln Gly Val Ile Arg Cys Val Ser Asn Ile Thr Gly Leu Ile Leu Thr 
                405                 410                 415     
Arg Asp Gly Gly Ser Thr Asp Ser Thr Thr Glu Thr Phe Arg Pro Ser 
            420                 425                 430         
Gly Gly Asp Met Arg Asp Asn Trp Arg Ser Glu Leu Tyr Lys Tyr Lys 
        435                 440                 445             
Val Val Lys Ile Glu Pro Leu Gly Val Ala Pro Thr Arg Cys Lys Arg 
    450                 455                 460                 
Arg Val Val Gly Arg Arg Arg Arg Arg Arg Ala Val Gly Ile Gly Ala 
465                 470                 475                 480 
Val Phe Leu Gly Phe Leu Gly Ala Ala Gly Ser Thr Met Gly Ala Ala 
                485                 490                 495     
Ser Met Thr Leu Thr Val Gln Ala Arg Asn Leu Leu Ser Gly Ile Val 
            500                 505                 510         
Gln Gln Gln Ser Asn Leu Leu Arg Ala Pro Glu Ala Gln Gln His Leu 
        515                 520                 525             
Leu Lys Leu Thr Val Trp Gly Ile Lys Gln Leu Gln Ala Arg Val Leu 
    530                 535                 540                 
Ala Val Glu Arg Tyr Leu Arg Asp Gln Gln Leu Leu Gly Ile Trp Gly 
545                 550                 555                 560 
Cys Ser Gly Lys Leu Ile Cys Cys Thr Asn Val Pro Trp Asn Ser Ser 
                565                 570                 575     
Trp Ser Asn Arg Asn Leu Ser Glu Ile Trp Asp Asn Met Thr Trp Leu 
            580                 585                 590         
Gln Trp Asp Lys Glu Ile Ser Asn Tyr Thr Gln Ile Ile Tyr Gly Leu 
        595                 600                 605             
Leu Glu Glu Ser Gln Asn Gln Gln Glu Lys Asn Glu Gln Asp Leu Leu 
    610                 615                 620                 
Ala Leu Asp 
625         

<210> 4
<211> 41
<212> DNA
<213> Artificial Sequence


<220> 
<223> forward primer

<400> 4
atcaatgaaa accaggggaa tgagtacagg ctgatcaatt g                         41


<210> 5
<211> 41
<212> DNA
<213> Artificial Sequence


<220> 
<223> reverse primer

<400> 5
caattgatca gcctgtactc attcccctgg ttttcattga t                         41


<210> 6
<211> 16
<212> PRT
<213> Artificial Sequence


<220> 
<223> His-tag sequence

<400> 6
Gly Ser Gly Ser Gly Gly Ser Gly His His His His His His His His 
1               5                   10                  15      



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


<220> 
<223> deletion

<400> 7
Arg Ser Asn Asn Ser Asn Lys 
1               5           

<210> 8
<211> 19
<212> PRT
<213> Artificial Sequence


<220> 
<223> example upper

<400> 8
Leu Leu Leu Thr Arg Asp Gly Gly Asn Ser Asn Asn Glu Ser Glu Ile 
1               5                   10                  15      
Phe Arg Pro 
            

<210> 9
<211> 21
<212> PRT
<213> Artificial Sequence


<220> 
<223> example lower

<400> 9
Leu Leu Leu Thr Arg Asp Gly Gly Ser Asn Arg Ser Glu Pro Glu Val 
1               5                   10                  15      
Glu Ile Phe Arg Pro 
            20      

<210> 10
<211> 9
<212> PRT
<213> Artificial Sequence


<220> 
<223> deletion 434-442

<400> 10
Ile Asn Met Trp Gln Lys Val Gly Lys 
1               5                   

<210> 11
<211> 10
<212> PRT
<213> Artificial Sequence


<220> 
<223> deletion 434-443

<400> 11
Ile Asn Met Trp Gln Lys Val Gly Lys Ala 
1               5                   10  

<210> 12
<211> 11
<212> PRT
<213> Artificial Sequence


<220> 
<223> deletion 434-444

<400> 12
Ile Asn Met Trp Gln Lys Val Gly Lys Ala Met 
1               5                   10      

<210> 13
<211> 14
<212> PRT
<213> Artificial Sequence


<220> 
<223> deletion 429-442

<400> 13
Arg Ile Lys Gln Ile Ile Asn Met Trp Gln Lys Val Gly Lys 
1               5                   10                  

<210> 14
<211> 13
<212> PRT
<213> Artificial Sequence


<220> 
<223> deletion 430-442

<400> 14
Ile Lys Gln Ile Ile Asn Met Trp Gln Lys Val Gly Lys 
1               5                   10              

<210> 15
<211> 12
<212> PRT
<213> Artificial Sequence


<220> 
<223> deletion 431-442

<400> 15
Lys Gln Ile Ile Asn Met Trp Gln Lys Val Gly Lys 
1               5                   10          

<210> 16
<211> 11
<212> PRT
<213> Artificial Sequence


<220> 
<223> deletion 422-442

<400> 16
Gln Ile Ile Asn Met Trp Gln Lys Val Gly Lys 
1               5                   10      

<210> 17
<211> 10
<212> PRT
<213> Artificial Sequence


<220> 
<223> deletion 433-442

<400> 17
Ile Ile Asn Met Trp Gln Lys Val Gly Lys 
1               5                   10  

<210> 18
<211> 7
<212> PRT
<213> Artificial Sequence


<220> 
<223> linker

<400> 18
Asn Gly Thr Gly Gly Ser Gly 
1               5           

