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Supplemental Figure 6. Comparative organization of the reconstructed cetartiodactyl and pecoran ancestral chromosomes with the genome as a reference. and orientation of syntenic fragments are visualized using the Evolution Highway comparative chromosome browser. Blue and pink colors represent orientation of blocks relative to the reference, with blue indicating the same orientation, and pink indicating the opposite orientation. Pink does not always indicate an inversion because the orientation of RACFs is randomly chosen during the reconstruction. The number within each block represents a chromosome number for a reconstructed ancestor, and a lower-case letter indicates a fragment of the chromosome. Light grey blocks indicate mammalian multispecies homologous synteny blocks (msHSBs). Diamonds indicate the position of EBRs, with light and dark green showing EBRs classified as or pecoran, respectively, using FISH data. Heatmaps show the density of ruminant, cetartiodactyl, and mammalian enhancers, and ruminant gene family expansions in windows of 100 Kbp along cattle chromosomes.

Cattle chr1 DESCHR_Cetartio DESCHR_Pecora 0 Cattle Ruminant Cetartiodactyl Mammal Expansions

1

1

1

1 1b 1a

Bases 60000 1

40000 1

1 4.0e+07 20000 1

1a 1b 1 0

1b 1

1

1

1

1 8.0e+07 1

1

1

1a 1 1c

1

1 1 1 classification 1

1 ? 1.2e+08 1 Ruminant

1

1

1a 1b

1a 1b

1b 1b 1b 1a 1 1b 1a 1b

1b 1 1b

1a Cattle chr2 DESCHR_Cetartio DESCHR_Pecora Mammal 0 Cattle Ruminant Cetartiodactyl Mammal Expansions

6

2

2 2 6 2 Bases

2 20000

2 10000

6 2 6 2 0 2

2 5e+07

2

6

2

2

2

2 2

2

classification 1e+08 6 2 Pecora

Ruminant

2

2

2

3a 2 2 Cattle chr3 DESCHR_Cetartio DESCHR_Pecora Mammal 0 Cattle Ruminant Cetartiodactyl Mammal Expansions

3

3

3 Bases

3 15000

3 10000 3.0e+07 3 5000 3 0

3

3

3a 3 6.0e+07 3

3

3

3

3

3

3

3 classification 9.0e+07

3 Pecora

3

3

3

3

6 1.2e+08 Cattle chr4 DESCHR_Cetartio DESCHR_Pecora Mammal 0 Cattle Ruminant Cetartiodactyl Mammal Expansions

8a

4 4 8a 4

8a 4

8a 4

8a 4

8a 4 8a 4 Bases 4 20000

4 15000 10000 3.0e+07 4 8a 4 5000 0

4

8a 4 8a 4

8a 4 4

8b 4 4 8b 4 8b 4

6.0e+07

8b 4

4

4

8a

8a

4

4 classification 9.0e+07 4 Ruminant 4

4

4

4 8b 4

4

4

1.2e+08 Cattle chr5 DESCHR_Cetartio DESCHR_Pecora Mammal 0 Cattle Ruminant Cetartiodactyl Mammal Expansions

5

5

5 9a

5

5 Bases 80000 60000 5 40000 3.0e+07 20000 5 9a 0

5

9a 5

6 5 6.0e+07

5

5 9a

5

5

5 classification 9.0e+07 Ruminant 9b 5

5

5

9a 5

9a 6

9a 6

9a 6 1.2e+08 9a 6 Cattle chr6 DESCHR_Cetartio DESCHR_Pecora Mammal 0 Cattle Ruminant Cetartiodactyl Mammal Expansions

4b

6

6

6 4a Bases

6 10000

6 3e+07 5000

6

6 0

6

6

5 6

6e+07

4a

6

6

classification

6 9e+07 Pecora

6

6

6

4c

4c 5 4c 5 4c 5

4c 5

6 4a 5 6

4c Cattle chr7 DESCHR_Cetartio DESCHR_Pecora Mammal 0 Cattle Ruminant Cetartiodactyl Mammal Expansions

2a

2c

7

2b

2b Bases 2b 15000

7 10000

3e+07 5000 2a 7 0

2a

2d

2b 7

8 7

7 6e+07

7 2a 7

7

7 classification 7 ?

7 Pecora

7 Ruminant 9e+07 7

7 2a

7

7 Cattle chr8 DESCHR_Cetartio DESCHR_Pecora Mammal 0 Cattle Ruminant Cetartiodactyl Mammal Expansions

25

8

25

25 8

8

8 Bases 8 15000

8 10000

3e+07 5000 8 0

8 11c

8

8

8

8 7 8 6e+07 8

8

25 8

8

8

11c classification

8 Pecora

11e Ruminant 9e+07 11d

8

8

8

11c 8

8

14 Cattle chr9 DESCHR_Cetartio DESCHR_Pecora Mammal 0 Cattle Ruminant Cetartiodactyl Mammal Expansions

9 10

9 Bases 100000

9 75000 2.5e+07 50000 9 25000 9 0

9

9

9

10 9

5.0e+07 10 9

9

9

9

7.5e+07 9 classification Pecora 9

9

10 9

9

9

1.0e+08

9 Cattle chr10 DESCHR_Cetartio DESCHR_Pecora Mammal 0 Cattle Ruminant Cetartiodactyl Mammal Expansions

2a

10

2a

2a 10

5

10

5 10 Bases 10 80000 10 60000 2.5e+07 40000 10 20000 5 0

10

10 5 11 10

10

5.0e+07 10

10 5 10

10

10

10

classification 10 7.5e+07 ? 5

10 Pecora

10 Ruminant

10

2a 8

10

10

5 11 10 1.0e+08

10 Cattle chr11 DESCHR_Cetartio DESCHR_Pecora Mammal 0 Cattle Ruminant Cetartiodactyl Mammal Expansions

14 11

11

14 11

11

11 Bases

11 20000 15000 2.5e+07 11 10000 14 5000 11 0

11

11

14

5.0e+07 11

14 9 11

11

11

14 11

11

11

7.5e+07 classification 11 ?

14 11 Pecora

11 Ruminant

11

11

11c 11

1.0e+08 11

11b 11c 11a 9 11c Cattle chr12 DESCHR_Cetartio DESCHR_Pecora Mammal 0 Cattle Ruminant Cetartiodactyl Mammal Expansions

12

Bases 13a 20000 12 15000 10000 5000 2.5e+07 12 0

12

12

13 12

5.0e+07

13a

12

12

classification 12 ?

7.5e+07 13b

12

12

13a 12 Cattle chr13 DESCHR_Cetartio DESCHR_Pecora Mammal 0 Cattle Ruminant Cetartiodactyl Mammal Expansions

19 17b

13

13 24b 17a Bases 13 15000 24a

2e+07 10000 5000 24a 13 0

24a

4e+07 19

24b 13

19 13 17a 13 19

19 13 6e+07 classification

13 ?

Ruminant

19

8e+07 Cattle chr14 DESCHR_Cetartio DESCHR_Pecora Mammal 0 Cattle Ruminant Cetartiodactyl Mammal Expansions

14

15 14

14

15 Bases 20000 15 14 15000 2e+07 10000

14 5000 0 14

15 14

14 4e+07 14 14

14

14

6e+07 classification Pecora 15 14

14

14

8e+07 14

15 Cattle chr15 DESCHR_Cetartio DESCHR_Pecora Mammal 0 Cattle Ruminant Cetartiodactyl Mammal Expansions

7a

15

7a

15

Bases

15 30000 2e+07 20000 15 10000

0 7b 15

15

15 4e+07 7a 15

7c

7a 15

6e+07 15 classification 15 Pecora

7a

15

15 8e+07

7b Cattle chr16 DESCHR_Cetartio DESCHR_Pecora Mammal 0 Cattle Ruminant Cetartiodactyl Mammal Expansions

16

3a 16

16 Bases

15000 10000 2e+07 3b 16 5000 0

16 16

16 3a 4e+07 16

3a 16

16

3b 16

3a 16

3a

16 classification 6e+07 16 Pecora

16 3a

16

16 16

8e+07 3b Cattle chr17 DESCHR_Cetartio DESCHR_Pecora Mammal 0 Cattle Ruminant Cetartiodactyl Mammal Expansions

17

17 Bases 5000 17 4000 3000 4a 2000 2e+07 1000 17 0

17

17

17

18 4e+07 4b 17

17

17

17

17 classification Ruminant

23a 17 6e+07

17

23b

23b Cattle chr18 DESCHR_Cetartio DESCHR_Pecora Mammal 0 Cattle Ruminant Cetartiodactyl Mammal Expansions

18b

18 18a

Bases 80000 18 60000 40000 20000 0 2e+07 18c 18

18a

18a 22 18

18

4e+07

18

classification ?

Pecora 18b Ruminant

18

6e+07

18d

18d Cattle chr19 DESCHR_Cetartio DESCHR_Pecora Mammal 0 Cattle Ruminant Cetartiodactyl Mammal Expansions

19

Bases 19 50000 40000 30000

19 20000 10000 0

2e+07 19

16

19 21

19

4e+07 19

19

classification

19 ?

19

16 19

6e+07 19 Cattle chr20 DESCHR_Cetartio DESCHR_Pecora Mammal 0 Cattle Ruminant Cetartiodactyl Mammal Expansions

2a

20

20

20 Bases 20000 20 15000 10000 5000 2e+07 20 0

20

20 19

20 4e+07 2a

20

20

classification 20 ?

20

20 6e+07

20

20 Cattle chr21 DESCHR_Cetartio DESCHR_Pecora Mammal 0 Cattle Ruminant Cetartiodactyl Mammal Expansions

5 20

21

21 5 Bases

21 75000 50000

21 25000 2e+07 0

5

5

5

5 20 5 21

4e+07 21

5 21

classification ?

Pecora

5 Ruminant

5

6e+07 22

21

27 5 Cattle chr22 DESCHR_Cetartio DESCHR_Pecora Mammal 0 Cattle Ruminant Cetartiodactyl Mammal Expansions

8a

22

24 1a 22

Bases

15000

1a 24 10000

1a 24

1a 5000 0

2e+07 22

22

22 1a

24 4e+07 22

classification ? 22 Ruminant

22

1a

1a

6e+07 1a Cattle chr23 DESCHR_Cetartio DESCHR_Pecora Mammal 0 Cattle Ruminant Cetartiodactyl Mammal Expansions

20

23

Bases 1e+07 23 30000

20000

10000

0 20 23

2e+07

26

3e+07

23

23

classification 20 ? 4e+07 23

23

5e+07 Cattle chr24 DESCHR_Cetartio DESCHR_Pecora Mammal 0 Cattle Ruminant Cetartiodactyl Mammal Expansions

17b

24

24 Bases

20000 15000 10000 24 5000 0

2e+07

24

24

17a 24 23

24

24

4e+07 24

24 classification Ruminant

24

17b 24

6e+07 Cattle chr25 DESCHR_Cetartio DESCHR_Pecora Mammal 0 Cattle Ruminant Cetartiodactyl Mammal Expansions

25

Bases 12500

25 10000 1e+07 7500 5000 2500 0

25 2e+07 21 28

25

3e+07 classification

25 ?

4e+07 Cattle chr26 DESCHR_Cetartio DESCHR_Pecora Mammal 0 Cattle Ruminant Cetartiodactyl Mammal Expansions

26

Bases 1e+07 50000 12b 12a 40000 30000 26 20000 10000 0

26 2e+07

12b 12a

3e+07

26

12b 12a classification ? 4e+07 26

26

26

5e+07

12b 12a 12b 12a Cattle chr27 DESCHR_Cetartio DESCHR_Pecora Mammal 0 Cattle Ruminant Cetartiodactyl Mammal Expansions

27 Bases

20000 1e+07 15000 27 10000 5000 0

27

27 2e+07 22

27 27

27 3e+07

classification ?

4e+07

1a Cattle chr28 DESCHR_Cetartio DESCHR_Pecora Mammal 0 Cattle Ruminant Cetartiodactyl Mammal Expansions 0e+00

12a 12b

12a 12a

28

Bases 12a 1e+07 28 10000

5000

0

28

2e+07

28 12a

28

3e+07 12b

28

classification Ruminant

28 4e+07 Cattle chr29 DESCHR_Cetartio DESCHR_Pecora Mammal 0 Cattle Ruminant Cetartiodactyl Mammal Expansions

29 Bases 1e+07 7a 80000 60000 29 40000 20000 0

29

2e+07

7a

29 25

3e+07 7b 29

7b 29

classification 7b ?

4e+07 Ruminant

29

7a

29

5e+07 Cattle chrX DESCHR_Cetartio DESCHR_Pecora Mammal 0 Cattle Ruminant Cetartiodactyl Mammal Expansions Xb Xh Xh

Xb

X Xb X Xb X

X Bases Xe X

Xe Xf X 20000 X Xe Xf 10000

Xh X 0 Xd Xe X 5e+07 Xd

Xg Xd X Xd Xg X Xf Xd Xf Xd X

Xb Xb Xi X Xb Xi

Xg Xc Xb Xg Xc X

X

Xc X X

Xa X

1e+08 Xc Xa X

X

Xa X classification ?

X Xa

X

X

Xa Xc Xa Xc Xa Xc Xa Xa Xc

Xa Xc