                         SEQUENCE LISTING

<110>  The United States of America, as represented by The 
       Secretary, Department of Health and Human Services
 
<120>  HUMAN INSULIN C-ALPHA-PEPTIDES AND METHODS OF USE

<130>  4239-105675-02

<150>  US 63/178,083
<151>  2021-04-22

<160>  71    

<170>  PatentIn version 3.5

<210>  1
<211>  204
<212>  PRT
<213>  Homo sapiens

<400>  1

Met Gly Ser Glu Thr Ile Lys Pro Ala Gly Ala Gln Gln Pro Ser Ala 
1               5                   10                  15      


Leu Gln Asp Arg Leu His Gln Lys Arg Pro Ser Ser Arg Ser Val Pro 
            20                  25                  30          


Arg Ala Phe Ala Ser Gly Gly Leu Arg Ile Pro Gly Trp Leu Asp Pro 
        35                  40                  45              


Arg Pro Gln Leu Cys Ser Arg Glu Asp Val Ala Gly Leu Val Lys His 
    50                  55                  60                  


Val Gly Val Ser Pro Gly Ala Pro Arg Gln Gly Thr Trp Pro Ser Ala 
65                  70                  75                  80  


Cys Leu Ser Pro Ala Cys Leu Pro Asp His Cys Pro Ser Ala Met Ala 
                85                  90                  95      


Leu Trp Met Arg Leu Leu Pro Leu Leu Ala Leu Leu Ala Leu Trp Gly 
            100                 105                 110         


Pro Asp Pro Ala Ala Ala Phe Val Asn Gln His Leu Cys Gly Ser His 
        115                 120                 125             


Leu Val Glu Ala Leu Tyr Leu Val Cys Gly Glu Arg Gly Phe Phe Tyr 
    130                 135                 140                 


Thr Pro Lys Thr Arg Arg Glu Ala Glu Asp Leu Gln Val Gly Gln Val 
145                 150                 155                 160 


Glu Leu Gly Gly Gly Pro Gly Ala Gly Ser Leu Gln Pro Leu Ala Leu 
                165                 170                 175     


Glu Gly Ser Leu Gln Lys Arg Gly Ile Val Glu Gln Cys Cys Thr Ser 
            180                 185                 190         


Ile Cys Ser Leu Tyr Gln Leu Glu Asn Tyr Cys Asn 
        195                 200                 


<210>  2
<211>  198
<212>  PRT
<213>  Homo sapiens

<400>  2

Met Gly Ser Glu Thr Ile Lys Pro Ala Gly Ala Gln Gln Pro Ser Ala 
1               5                   10                  15      


Leu Gln Asp Arg Leu His Gln Lys Arg Pro Ser Ser Arg Ser Val Pro 
            20                  25                  30          


Arg Ala Phe Ala Ser Gly Gly Leu Arg Ile Pro Gly Trp Leu Asp Pro 
        35                  40                  45              


Arg Pro Gln Leu Cys Ser Arg Glu Asp Val Ala Gly Leu Val Lys His 
    50                  55                  60                  


Val Gly Val Ser Pro Gly Ala Pro Arg Gln Gly Thr Trp Pro Ser Ala 
65                  70                  75                  80  


Cys Leu Ser Pro Ala Cys Leu Pro Asp His Cys Pro Ser Ala Met Ala 
                85                  90                  95      


Leu Trp Met Arg Leu Leu Pro Leu Leu Ala Leu Leu Ala Leu Trp Gly 
            100                 105                 110         


Pro Asp Pro Ala Ala Ala Phe Val Asn Gln His Leu Cys Gly Ser His 
        115                 120                 125             


Leu Val Glu Ala Leu Tyr Leu Val Cys Gly Glu Arg Gly Phe Phe Tyr 
    130                 135                 140                 


Thr Pro Lys Thr Arg Arg Glu Ala Glu Asp Leu Gln Gly Glu Pro Thr 
145                 150                 155                 160 


Ala His Cys Cys Pro Trp Pro Pro Pro Ala Thr Pro Cys Ser Trp Arg 
                165                 170                 175     


Ser His Pro Ala Trp Ala Glu Gly Gly Arg Arg Leu Pro Pro Ser Arg 
            180                 185                 190         


Gly Ser Gly Ala Leu Phe 
        195             


<210>  3
<211>  53
<212>  PRT
<213>  Homo sapiens

<400>  3

Met Gly Ser Glu Thr Ile Lys Pro Ala Gly Ala Gln Gln Pro Ser Ala 
1               5                   10                  15      


Leu Gln Asp Arg Leu His Gln Lys Arg Pro Ser Ser Arg Ser Leu Ser 
            20                  25                  30          


Phe Cys His Gly Pro Val Asp Ala Pro Pro Ala Pro Ala Gly Ala Ala 
        35                  40                  45              


Gly Pro Leu Gly Thr 
    50              


<210>  4
<211>  153
<212>  PRT
<213>  Homo sapiens

<400>  4

Met Gly Ser Glu Thr Ile Lys Pro Ala Gly Ala Gln Gln Pro Ser Ala 
1               5                   10                  15      


Leu Gln Asp Arg Leu His Gln Lys Arg Pro Ser Ser Arg Ser Val Pro 
            20                  25                  30          


Arg Ala Phe Ala Ser Asp His Cys Pro Ser Ala Met Ala Leu Trp Met 
        35                  40                  45              


Arg Leu Leu Pro Leu Leu Ala Leu Leu Ala Leu Trp Gly Pro Asp Pro 
    50                  55                  60                  


Ala Ala Ala Phe Val Asn Gln His Leu Cys Gly Ser His Leu Val Glu 
65                  70                  75                  80  


Ala Leu Tyr Leu Val Cys Gly Glu Arg Gly Phe Phe Tyr Thr Pro Lys 
                85                  90                  95      


Thr Arg Arg Glu Ala Glu Asp Leu Gln Val Gly Gln Val Glu Leu Gly 
            100                 105                 110         


Gly Gly Pro Gly Ala Gly Ser Leu Gln Pro Leu Ala Leu Glu Gly Ser 
        115                 120                 125             


Leu Gln Lys Arg Gly Ile Val Glu Gln Cys Cys Thr Ser Ile Cys Ser 
    130                 135                 140                 


Leu Tyr Gln Leu Glu Asn Tyr Cys Asn 
145                 150             


<210>  5
<211>  73
<212>  PRT
<213>  Homo sapiens

<400>  5

Met Gly Ser Glu Thr Ile Lys Pro Ala Gly Ala Gln Gln Pro Ser Ala 
1               5                   10                  15      


Leu Gln Asp Arg Leu His Gln Lys Arg Pro Ser Ser Arg Ser Val Pro 
            20                  25                  30          


Arg Ala Phe Ala Ser Gly Gly Leu Arg Ile Pro Gly Trp Leu Asp Pro 
        35                  40                  45              


Arg Ser Leu Ser Phe Cys His Gly Pro Val Asp Ala Pro Pro Ala Pro 
    50                  55                  60                  


Ala Gly Ala Ala Gly Pro Leu Gly Thr 
65                  70              


<210>  6
<211>  98
<212>  PRT
<213>  Homo sapiens

<400>  6

Met Ala Leu Trp Met Arg Leu Leu Pro Leu Leu Ala Leu Leu Ala Leu 
1               5                   10                  15      


Trp Gly Pro Asp Pro Ala Ala Ala Phe Val Asn Gln His Leu Cys Gly 
            20                  25                  30          


Ser His Leu Val Glu Ala Leu Tyr Leu Val Cys Gly Glu Arg Gly Phe 
        35                  40                  45              


Phe Tyr Thr Pro Lys Thr Arg Arg Glu Ala Glu Asp Leu Gln Gly Ser 
    50                  55                  60                  


Leu Gln Pro Leu Ala Leu Glu Gly Ser Leu Gln Lys Arg Gly Ile Val 
65                  70                  75                  80  


Glu Gln Cys Cys Thr Ser Ile Cys Ser Leu Tyr Gln Leu Glu Asn Tyr 
                85                  90                  95      


Cys Asn 
        


<210>  7
<211>  19
<212>  PRT
<213>  Homo sapiens

<400>  7

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


Ser Leu Gln 
            


<210>  8
<211>  20
<212>  PRT
<213>  Homo sapiens

<400>  8

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


Ser Leu Gln Lys 
            20  


<210>  9
<211>  18
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS EX2 probe

<400>  9
tgaaccaaca cctgtgcg                                                     18


<210>  10
<211>  16
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS E2 forward primer

<400>  10
cccagccgca gccttt                                                       16


<210>  11
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS EX2 reverse primer

<400>  11
agagagcttc caccaggtgt ga                                                22


<210>  12
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS EX2-12 probe

<400>  12
acgaggcttc ttctacacac                                                   20


<210>  13
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS EX2-12 forward primer

<400>  13
tcacacctgg tggaagctct ct                                                22


<210>  14
<211>  18
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS EX2-12 reverse primer

<400>  14
ggcagcaatg ggcagttg                                                     18


<210>  15
<211>  19
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS INS1A probe

<400>  15
ccatcaagca gatcactgt                                                    19


<210>  16
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS INS1A forward primer

<400>  16
ggacaggctg catcagaaga g                                                 21


<210>  17
<211>  18
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS INS1A reverse primer

<400>  17
acagggccat ggcagaag                                                     18


<210>  18
<211>  16
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS INS1B probe

<400>  18
tttgcgtcag atcact                                                       16


<210>  19
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS INS1B forward  primer

<400>  19
ccatcaagca ggtctgttcc a                                                 21


<210>  20
<211>  17
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS INS1B reverse primer

<400>  20
gggccatggc agaagga                                                      17


<210>  21
<211>  16
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS INS1C probe

<400>  21
accccagatc actgtc                                                       16


<210>  22
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS INS1C forward primer

<400>  22
ccatcaagca ggtctgttcc a                                                 21


<210>  23
<211>  16
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS INS1C reverse primer

<400>  23
gcaggaggcg catcca                                                       16


<210>  24
<211>  16
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS INSI1 priobe

<400>  24
ccatcaagca ggtctg                                                       16


<210>  25
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS INSI1 forward primer

<400>  25
ggacaggctg catcagaaga g                                                 21


<210>  26
<211>  18
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS INSI1 reverse primer

<400>  26
agcccacctg acgcaaag                                                     18


<210>  27
<211>  19
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS INSUA probe

<400>  27
ccatcaagca gatcactgt                                                    19


<210>  28
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS INSUA forward primer

<400>  28
gggagatggg ctctgagact ataa                                              24


<210>  29
<211>  18
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS INSUA reverse primer

<400>  29
acagggccat ggcagaag                                                     18


<210>  30
<211>  16
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS INSUB probe

<400>  30
tttgcgtcag atcact                                                       16


<210>  31
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS INSUB forward primer

<400>  31
gggagatggg ctctgagact ataa                                              24


<210>  32
<211>  17
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS INSUB reverse primer

<400>  32
gggccatggc agaagga                                                      17


<210>  33
<211>  16
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS INSUC probe

<400>  33
accccagatc actgtc                                                       16


<210>  34
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS INSUC forward primer

<400>  34
gggagatggg ctctgagact ataa                                              24


<210>  35
<211>  16
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS INSUC reverse primer

<400>  35
gcaggaggcg catcca                                                       16


<210>  36
<211>  16
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS INSU1 probe

<400>  36
ccatcaagca ggtctg                                                       16


<210>  37
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS INSU1 forward primer

<400>  37
gggagatggg ctctgagact ataa                                              24


<210>  38
<211>  18
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS INSU1 reverse primer

<400>  38
agcccacctg acgcaaag                                                     18


<210>  39
<211>  15
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS INS3B probe

<400>  39
caggctgccc tgcag                                                        15


<210>  40
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS INS3B forward primer

<400>  40
tagagggagc agatgctggt aca                                               23


<210>  41
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  INS INS3B reverse primer

<400>  41
aggcttcttc tacacaccca aga                                               23


<210>  42
<211>  19
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Ins Ins2-V1 probe

<400>  42
caagcaggaa ggttattgt                                                    19


<210>  43
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Ins Ins2-V1 forward primer

<400>  43
ccgctacaat caaaaaccat ca                                                22


<210>  44
<211>  17
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Ins Ins2-V1 reverse primer

<400>  44
catccacagg gccatgt                                                      17


<210>  45
<211>  17
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Ins Ins2-V2 probe

<400>  45
caagcaggaa ggtactc                                                      17


<210>  46
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Ins Ins2-V2 forward primer

<400>  46
ccgctacaat caaaaaccat ca                                                22


<210>  47
<211>  18
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Ins Ins2-V2 reverse primer

<400>  47
gagccaggcc cactgaga                                                     18


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

<220>
<223>  Ins Ins2-V3 probe

<400>  48
atccgctaca atcaa                                                        15


<210>  49
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Ins Ins2-V3 forward primer

<400>  49
ccagtaacca ccagccctaa gt                                                22


<210>  50
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Ins Ins2-V3 reverse primer

<400>  50
agataggctt cctgcttgct gatg                                              24


<210>  51
<211>  19
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Ins Ins2-V4 probe

<400>  51
ctggaggtta ttgtttcaa                                                    19


<210>  52
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Ins Ins2-V4 forward primer

<400>  52
gctcctacgc tgaaattcca a                                                 21


<210>  53
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Ins Ins2-V4 reverse primer

<400>  53
aaggtgctgc ttgacaaaag c                                                 21


<210>  54
<211>  22
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Insulin tryptic peptide pep-B

<400>  54

Phe Val Asn Gln His Leu Cys Gly Ser His Leu Val Glu Ala Leu Tyr 
1               5                   10                  15      


Leu Val Cys Gly Glu Arg 
            20          


<210>  55
<211>  21
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Insulin tryptic peptide pep-A

<400>  55

Gly Ile Val Glu Gln Cys Cys Thr Ser Ile Cys Ser Leu Tyr Gln Leu 
1               5                   10                  15      


Glu Asn Tyr Cys Asn 
            20      


<210>  56
<211>  19
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Insulin tryptic peptide pep-Calpha

<400>  56

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


Ser Leu Gln 
            


<210>  57
<211>  20
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Insulin tryptic peptide pep-CalphaK

<400>  57

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


Ser Leu Gln Lys 
            20  


<210>  58
<211>  13
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Insulin tryptic peptide pep-U1

<400>  58

Pro Ala Gly Ala Gln Gln Pro Ser Ala Leu Gln Asp Arg 
1               5                   10              


<210>  59
<211>  8
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Insulin tryptic peptide pep-U2

<400>  59

Ala Phe Ala Ser Gly Gly Leu Arg 
1               5               


<210>  60
<211>  8
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Insulin tryptic peptide pep-U3

<400>  60

Ile Pro Gly Trp Leu Asp Pro Arg 
1               5               


<210>  61
<211>  8
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Insulin tryptic peptide pep-U4

<400>  61

Glu Asp Val Ala Gly Leu Val Lys 
1               5               


<210>  62
<211>  10
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Insulin tryptic peptide pep-U5

<400>  62

His Val Gly Val Ser Pro Gly Ala Pro Arg 
1               5                   10  


<210>  63
<211>  16
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Insulin tryptic peptide pep-US

<400>  63

Ala Phe Ala Ser Asp His Cys Pro Ser Ala Met Ala Leu Trp Met Arg 
1               5                   10                  15      


<210>  64
<211>  10
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Insulin tryptic peptide pep-UF

<400>  64

Ser His Pro Ala Trp Ala Glu Gly Gly Arg 
1               5                   10  


<210>  65
<211>  12
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Alternative consensus Kozak ribosomal binding site

<400>  65
tgggagatgg gc                                                           12


<210>  66
<211>  19
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  C-alpha peptide

<400>  66

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


Ser Leu Gln 
            


<210>  67
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Beta sheet forming motif

<400>  67

Gly Gln Val Glu Leu 
1               5   


<210>  68
<211>  14
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Instrinsically disordered protein region sequence

<400>  68

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


<210>  69
<211>  110
<212>  PRT
<213>  Homo sapiens

<400>  69

Met Ala Leu Trp Met Arg Leu Leu Pro Leu Leu Ala Leu Leu Ala Leu 
1               5                   10                  15      


Trp Gly Pro Asp Pro Ala Ala Ala Phe Val Asn Gln His Leu Cys Gly 
            20                  25                  30          


Ser His Leu Val Glu Ala Leu Tyr Leu Val Cys Gly Glu Arg Gly Phe 
        35                  40                  45              


Phe Tyr Thr Pro Lys Thr Arg Arg Glu Ala Glu Asp Leu Gln Val Gly 
    50                  55                  60                  


Gln Val Glu Leu Gly Gly Gly Pro Gly Ala Gly Ser Leu Gln Pro Leu 
65                  70                  75                  80  


Ala Leu Glu Gly Ser Leu Gln Lys Arg Gly Ile Val Glu Gln Cys Cys 
                85                  90                  95      


Thr Ser Ile Cys Ser Leu Tyr Gln Leu Glu Asn Tyr Cys Asn 
            100                 105                 110 


<210>  70
<211>  10
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Human serum albumin tryptic peptide

<400>  70

Leu Val Asn Glu Val Thr Glu Phe Ala Lys 
1               5                   10  


<210>  71
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Insulin tryptic peptide

<400>  71

Gly Phe Phe Tyr Thr Pro Lys 
1               5           


