                         SEQUENCE LISTING

<110>  University of Maryland Baltimore County
       Ostrand-Rosenberg, Suzanne
       Carter, Darryl L.
 
<120>  A RECOMBINANT BI-SPECIFIC POLYPEPTIDE FOR COORDINATELY ACTIVATING
        TUMOR-REACTIVE T CELLS AND NEUTRALIZING IMMUNE SUPPRESSION

<130>  026228.392PCT

<150>  62/268,682
<151>  2015-12-17

<150>  62/355,422
<151>  2016-06-28

<160>  16    

<170>  PatentIn version 3.5

<210>  1
<211>  318
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic construct

<400>  1
gaaattgtgt tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc       60

ctctcctgca gggccagtca gagtgttagc agctacttag cctggtacca acagaaacct      120

ggccaggctc ccaggctcct catctatgat gcatccaaca gggccactgg catcccagcc      180

aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag cctagagcct      240

gaagattttg cagtttatta ctgtcagcag cgtagcaact ggccgacgtt cggccaaggg      300

accaaggtgg aaatcaaa                                                    318


<210>  2
<211>  369
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic construct

<400>  2
caggtccaac tggtgcagtc tggggctgag gtcaagaagc ctgggtcgtc ggtgaaggtc       60

tcctgcaaga cttctggaga caccttcagc acctatgcta tcagctgggt gcgacaggcc      120

cctggacaag ggcttgagtg gatgggaggg atcatcccta tatttggtaa agcacactac      180

gcacagaagt tccagggcag agtcacgatt accgcggacg aatccacgag cacagcctac      240

atggagctga gcagcctgag atctgaggac acggccgtgt atttttgtgc gagaaagttt      300

cactttgttt cggggagccc cttcggtatg gacgtctggg gccaagggac cacggtcacc      360

gtctcctca                                                              369


<210>  3
<211>  318
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic construct

<400>  3
gacattcagc tgacccagtc tccagcaatc atgtctgcat ctccagggga gaaggtcacc       60

atgacctgca gagccagttc aagtgtaagt tacatgaact ggtaccagca gaagtcaggc      120

acctccccca aaagatggat ttatgacaca tccaaagtgg cttctggagt cccttatcgc      180

ttcagtggca gtgggtctgg gacctcatac tctctcacaa tcagcagcat ggaggctgaa      240

gatgctgcca cttattactg ccaacagtgg agtagtaacc cgctcacgtt cggtgctggg      300

accaagctgg agctgaaa                                                    318


<210>  4
<211>  363
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic construct

<400>  4
gatatcaaac tgcagcagtc aggggctgaa ctggcaagac ctggggcctc agtgaagatg       60

tcctgcaaga cttctggcta cacctttact aggtacacga tgcactgggt aaaacagagg      120

cctggacagg gtctggaatg gattggatac attaatccta gccgtggtta tactaattac      180

aatcagaagt tcaaggacaa ggccacattg actacagaca aatcctccag cacagcctac      240

atgcaactga gcagcctgac atctgaggac tctgcagtct attactgtgc aagatattat      300

gatgatcatt actgccttga ctactggggc caaggcacca ctctcacagt ctcctcagtc      360

gaa                                                                    363


<210>  5
<211>  106
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic construct

<400>  5

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 
1               5                   10                  15      


Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr 
            20                  25                  30          


Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 
        35                  40                  45              


Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 
    50                  55                  60                  


Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro 
65                  70                  75                  80  


Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Thr 
                85                  90                  95      


Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 
            100                 105     


<210>  6
<211>  123
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic construct

<400>  6

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 
1               5                   10                  15      


Ser Val Lys Val Ser Cys Lys Thr Ser Gly Asp Thr Phe Ser Thr Tyr 
            20                  25                  30          


Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 
        35                  40                  45              


Gly Gly Ile Ile Pro Ile Phe Gly Lys Ala His Tyr Ala Gln Lys Phe 
    50                  55                  60                  


Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr 
65                  70                  75                  80  


Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Phe Cys 
                85                  90                  95      


Ala Arg Lys Phe His Phe Val Ser Gly Ser Pro Phe Gly Met Asp Val 
            100                 105                 110         


Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 
        115                 120             


<210>  7
<211>  106
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic construct

<400>  7

Asp Ile Gln Leu Thr Gln Ser Pro Ala Ile Met Ser Ala Ser Pro Gly 
1               5                   10                  15      


Glu Lys Val Thr Met Thr Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 
            20                  25                  30          


Asn Trp Tyr Gln Gln Lys Ser Gly Thr Ser Pro Lys Arg Trp Ile Tyr 
        35                  40                  45              


Asp Thr Ser Lys Val Ala Ser Gly Val Pro Tyr Arg Phe Ser Gly Ser 
    50                  55                  60                  


Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Glu Ala Glu 
65                  70                  75                  80  


Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Leu Thr 
                85                  90                  95      


Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 
            100                 105     


<210>  8
<211>  121
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic construct

<400>  8

Asp Ile Lys Leu Gln Gln Ser Gly Ala Glu Leu Ala Arg Pro Gly Ala 
1               5                   10                  15      


Ser Val Lys Met Ser Cys Lys Thr Ser Gly Tyr Thr Phe Thr Arg Tyr 
            20                  25                  30          


Thr Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 
        35                  40                  45              


Gly Tyr Ile Asn Pro Ser Arg Gly Tyr Thr Asn Tyr Asn Gln Lys Phe 
    50                  55                  60                  


Lys Asp Lys Ala Thr Leu Thr Thr Asp Lys Ser Ser Ser Thr Ala Tyr 
65                  70                  75                  80  


Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 
                85                  90                  95      


Ala Arg Tyr Tyr Asp Asp His Tyr Cys Leu Asp Tyr Trp Gly Gln Gly 
            100                 105                 110         


Thr Thr Leu Thr Val Ser Ser Val Glu 
        115                 120     


<210>  9
<211>  1533
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic construct

<400>  9
atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt ccactccgaa       60

attgtgttga cacagtctcc agccaccctg tctttgtctc caggggaaag agccaccctc      120

tcctgcaggg ccagtcagag tgttagcagc tacttagcct ggtaccaaca gaaacctggc      180

caggctccca ggctcctcat ctatgatgca tccaacaggg ccactggcat cccagccagg      240

ttcagtggca gtgggtctgg gacagacttc actctcacca tcagcagcct agagcctgaa      300

gattttgcag tttattactg tcagcagcgt agcaactggc cgacgttcgg ccaagggacc      360

aaggtggaaa tcaaaggtgg aggtggatct gggggaggtg gatctggggg tgggggatcg      420

caggtccaac tggtgcagtc tggggctgag gtcaagaagc ctgggtcgtc ggtgaaggtc      480

tcctgcaaga cttctggaga caccttcagc acctatgcta tcagctgggt gcgacaggcc      540

cctggacaag ggcttgagtg gatgggaggg atcatcccta tatttggtaa agcacactac      600

gcacagaagt tccagggcag agtcacgatt accgcggacg aatccacgag cacagcctac      660

atggagctga gcagcctgag atctgaggac acggccgtgt atttttgtgc gagaaagttt      720

cactttgttt cggggagccc cttcggtatg gacgtctggg gccaagggac cacggtcacc      780

gtctcctcag gaggtggtgg atccgatatc aaactgcagc agtcaggggc tgaactggca      840

agacctgggg cctcagtgaa gatgtcctgc aagacttctg gctacacctt tactaggtac      900

acgatgcact gggtaaaaca gaggcctgga cagggtctgg aatggattgg atacattaat      960

cctagccgtg gttatactaa ttacaatcag aagttcaagg acaaggccac attgactaca     1020

gacaaatcct ccagcacagc ctacatgcaa ctgagcagcc tgacatctga ggactctgca     1080

gtctattact gtgcaagata ttatgatgat cattactgcc ttgactactg gggccaaggc     1140

accactctca cagtctcctc agtcgaaggt ggaagtggag gttctggtgg aagtggaggt     1200

tcaggtggag tcgacgacat tcagctgacc cagtctccag caatcatgtc tgcatctcca     1260

ggggagaagg tcaccatgac ctgcagagcc agttcaagtg taagttacat gaactggtac     1320

cagcagaagt caggcacctc ccccaaaaga tggatttatg acacatccaa agtggcttct     1380

ggagtccctt atcgcttcag tggcagtggg tctgggacct catactctct cacaatcagc     1440

agcatggagg ctgaagatgc tgccacttat tactgccaac agtggagtag taacccgctc     1500

acgttcggtg ctgggaccaa gctggagctg aaa                                  1533


<210>  10
<211>  492
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic construct

<400>  10

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 
1               5                   10                  15      


Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr 
            20                  25                  30          


Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 
        35                  40                  45              


Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 
    50                  55                  60                  


Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro 
65                  70                  75                  80  


Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Thr 
                85                  90                  95      


Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser Gly 
            100                 105                 110         


Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Val Gln Ser 
        115                 120                 125             


Gly Ala Glu Val Lys Lys Pro Gly Ser Ser Val Lys Val Ser Cys Lys 
    130                 135                 140                 


Thr Ser Gly Asp Thr Phe Ser Thr Tyr Ala Ile Ser Trp Val Arg Gln 
145                 150                 155                 160 


Ala Pro Gly Gln Gly Leu Glu Trp Met Gly Gly Ile Ile Pro Ile Phe 
                165                 170                 175     


Gly Lys Ala His Tyr Ala Gln Lys Phe Gln Gly Arg Val Thr Ile Thr 
            180                 185                 190         


Ala Asp Glu Ser Thr Ser Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg 
        195                 200                 205             


Ser Glu Asp Thr Ala Val Tyr Phe Cys Ala Arg Lys Phe His Phe Val 
    210                 215                 220                 


Ser Gly Ser Pro Phe Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val 
225                 230                 235                 240 


Thr Val Ser Ser Gly Gly Gly Gly Ser Asp Ile Lys Leu Gln Gln Ser 
                245                 250                 255     


Gly Ala Glu Leu Ala Arg Pro Gly Ala Ser Val Lys Met Ser Cys Lys 
            260                 265                 270         


Thr Ser Gly Tyr Thr Phe Thr Arg Tyr Thr Met His Trp Val Lys Gln 
        275                 280                 285             


Arg Pro Gly Gln Gly Leu Glu Trp Ile Gly Tyr Ile Asn Pro Ser Arg 
    290                 295                 300                 


Gly Tyr Thr Asn Tyr Asn Gln Lys Phe Lys Asp Lys Ala Thr Leu Thr 
305                 310                 315                 320 


Thr Asp Lys Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr 
                325                 330                 335     


Ser Glu Asp Ser Ala Val Tyr Tyr Cys Ala Arg Tyr Tyr Asp Asp His 
            340                 345                 350         


Tyr Cys Leu Asp Tyr Trp Gly Gln Gly Thr Thr Leu Thr Val Ser Ser 
        355                 360                 365             


Val Glu Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly 
    370                 375                 380                 


Val Asp Asp Ile Gln Leu Thr Gln Ser Pro Ala Ile Met Ser Ala Ser 
385                 390                 395                 400 


Pro Gly Glu Lys Val Thr Met Thr Cys Arg Ala Ser Ser Ser Val Ser 
                405                 410                 415     


Tyr Met Asn Trp Tyr Gln Gln Lys Ser Gly Thr Ser Pro Lys Arg Trp 
            420                 425                 430         


Ile Tyr Asp Thr Ser Lys Val Ala Ser Gly Val Pro Tyr Arg Phe Ser 
        435                 440                 445             


Gly Ser Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Glu 
    450                 455                 460                 


Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro 
465                 470                 475                 480 


Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 
                485                 490         


<210>  11
<211>  45
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic construct

<400>  11
ggtggaggtg gatctggggg aggtggatct gggggtgggg gatcg                       45


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

<220>
<223>  Synthetic Construct

<400>  12

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 
1               5                   10                  15  


<210>  13
<211>  15
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic construct

<400>  13
ggaggtggtg gatcc                                                        15


<210>  14
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic construct

<400>  14

Gly Gly Gly Gly Ser 
1               5   


<210>  15
<211>  48
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic construct

<400>  15
ggtggaagtg gaggttctgg tggaagtgga ggttcaggtg gagtcgac                    48


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

<220>
<223>  Synthetic construct

<400>  16

Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly Val Asp 
1               5                   10                  15      


