1.

Supplementary Data

Table 1: Results of BLAST analysis.

For each XS candidate the suggested ortholog is listed, including the query cover and identity in percentage, as
well as the estimated E-Value on the day of the BLAST search (02.11.2020).

Nr. Candidate Ortholog Ortholog Ortholog E- Query Identity

Accession nr. Accession nr. Organism Annotation Value Cover [%]
[%]
1 ADI63839.1 WP_016951675.1 Anabaena sp. hypothetical 1e-55 100.0 81.98
PCC 7108 protein
2 BAL35925.1 NJK47103.1 Candidatus hypothetical 1e-65 97.0 80.00
Gracilibacteria protein
bacterium HC931_01810
3 ALJ68705.1 WP_162329005.1 Synechocystis hypothetical 8e-84 100.0 98.41
sp. CACIAM 05 protein
4 ACK73106.1 WP_190489364.1 Microcoleus sp. DUF3007 5e-49 96.0 72.00
FACHB-831 family protein
5 ABA24694.1 WP_158647785.1 Trichormus hypothetical 3e-53 72.0 100.00
variabilis protein
6 ALF53325.1 OCQ89361.1 Nostoc sp. hypothetical 3e-69 100.0 96.26
MBR 210 protein
BCD64_26765
7 BAK50588.1 NJK47103.1 Candidatus hypothetical 1e-65 97.0 80.00
Gracilibacteria protein
bacterium HC931_01810
8 AFY49197.1 WP_096552636.1 Nostoc sp. hypothetical 4e-73 100.0 90.00
NIES-4103 protein
Figure 1: Alignment of cyanobacterial XS candidates with Clustal Omega
A: PCR fragments of the candidates sll0911 and slr0582 were amplified from genomic Synechocystis DNA and
are 407 bp and 401 bp in size, respectively.
B: Fragments are shown at ca. 5711 bp for pET16b and 5493 bp for pET22b(+).
All samples shown were digested with the restriction enzymes NdeI and XhoI then run on a 1% agarose gel
with a 1 kb plus DNA Marker.
Table 2: General and bioinformatical information about the eight cyano-XS candidates selected for further analyses. Listed are the protein and DNA lengths of each
candidate, the start-codons, which information can be found up- and downstream of the target sequence, as well as which known domain motifs were found. Upstream
genes are toward the 5’ – end and downstream genes are toward the 3’ – end.

Nr. Candidates Protein DNA Start Upstream on same strand Downstream on same strand Domain motifs
length [aa] length [bp] codon
1 Nostoc azollae 0708 111 336 GTG - 2 hypothetical proteins - 4 hypothetical proteins none
- ABC transporter related
Protein
2 Synechocystis sp. PCC 128 387 ATG - ABC transporter - 2 hypothetical proteins - Rh5
6803 substr. PCC-P - RBD-FIP
3 Synechocystis sp. PCC 126 381 ATG - Hypothetical protein - GDP-L-fucose synthase - RBP_receptor
6803 - aluminum resistance family protein - 50S ribosomal protein L33 - ABC2_membrane_2
- 30S ribosomal protein S18
- alanine racemase
4 Gloeothece citriformis PCC 104 315 ATG - tryptophan synthase, alpha subunit - NADH dehydrogenase - DUF3007
7424 - ABC-3 protein subunit - Mis12
- class II aldolase/adducin - Tropomodulin
family protein - LapA_dom
- DUF1476
RPAP1_N
- RIC3
- UNC-93
5 Trichormus variabilis 114 345 GTG - hypothetical protein - 6 hypothetical proteins none
ATCC 29413 - Phage integrase
6 Nostoc piscinale CENA21 107 324 ATG - Pseudo gene - Pseudo protein - HTH_38
- chemotaxis protein CheY - ABC transporter substrate-
binding protein
7 Synechocystis sp. PCC 128 387 ATG - ABC-transporter - 2 hypothetical proteins - Rh5
6803 - RBD-FIP
8 Nostoc sp. PCC 7524 120 363 ATG - methylase involved in - Hypothetical protein - Carbam_trans_N
ubiquinone/menaquinone biosynthesis - pseudo gene - CRPA
- ring-hydroxylating dioxygenase, large - dihydroorotase-like cyclic
terminal subunit amidohydrolase
- hypothetical protein
- putative permease
Figure 2: PCR of the putative XS genes from Synechocystis and the pET-vectors
A: PCR fragments of the candidates sll0911 and slr0582 were amplified from genomic Synechocystis DNA and are 407 bp and 401 bp in size, respectively.
B: Fragments are shown at ca. 5711 bp for pET16b and 5493 bp for pET22b(+).
All samples shown were digested with the restriction enzymes NdeI and XhoI then run on a 1% agarose gel with a 1 kb plus DNA Marker.
Figure 3: Plasmid maps of the pET-vectors used for cloning
A: PCR fragments of the candidates sll0911 and slr0582 were amplified from genomic Synechocystis DNA and are 407 bp and 401 bp in size, respectively.
B: Fragments are shown at ca. 5711 bp for pET16b and 5493 bp for pET22b(+).
All samples shown were digested with the restriction enzymes NdeI and XhoI then run on a 1% agarose gel with a 1 kb plus DNA Marker.
Figure 4: PCR of the putative XS genes from Synechocystis and the pET-vectors
A: PCR fragments of the candidates sll0911 and slr0582 were amplified from genomic Synechocystis DNA and
are 407 bp and 401 bp in size, respectively.
B: Fragments are shown at ca. 5711 bp for pET16b and 5493 bp for pET22b(+).
All samples shown were digested with the restriction enzymes NdeI and XhoI then run on a 1% agarose gel
with a 1 kb plus DNA Marker.
Figure 5: PCR of the putative XS genes from Synechocystis and the pET-vectors
A: PCR fragments of the candidates sll0911 and slr0582 were amplified from genomic Synechocystis DNA and
are 407 bp and 401 bp in size, respectively.
B: Fragments are shown at ca. 5711 bp for pET16b and 5493 bp for pET22b(+).
All samples shown were digested with the restriction enzymes NdeI and XhoI then run on a 1% agarose gel
with a 1 kb plus DNA Marker.
Figure 6: PCR of the putative XS genes from Synechocystis and the pET-vectors
A: PCR fragments of the candidates sll0911 and slr0582 were amplified from genomic Synechocystis DNA and
are 407 bp and 401 bp in size, respectively.
B: Fragments are shown at ca. 5711 bp for pET16b and 5493 bp for pET22b(+).
All samples shown were digested with the restriction enzymes NdeI and XhoI then run on a 1% agarose gel
with a 1 kb plus DNA Marker.
Figure 7: PCR of the putative XS genes from Synechocystis and the pET-vectors
A: PCR fragments of the candidates sll0911 and slr0582 were amplified from genomic Synechocystis DNA and
are 407 bp and 401 bp in size, respectively.
B: Fragments are shown at ca. 5711 bp for pET16b and 5493 bp for pET22b(+).
All samples shown were digested with the restriction enzymes NdeI and XhoI then run on a 1% agarose gel
with a 1 kb plus DNA Marker.
Figure 8: PCR of the putative XS genes from Synechocystis and the pET-vectors
A: PCR fragments of the candidates sll0911 and slr0582 were amplified from genomic Synechocystis DNA and
are 407 bp and 401 bp in size, respectively.
B: Fragments are shown at ca. 5711 bp for pET16b and 5493 bp for pET22b(+).
All samples shown were digested with the restriction enzymes NdeI and XhoI then run on a 1% agarose gel
with a 1 kb plus DNA Marker.
Figure 9: PCR of the putative XS genes from Synechocystis and the pET-vectors
A: PCR fragments of the candidates sll0911 and slr0582 were amplified from genomic Synechocystis DNA and
are 407 bp and 401 bp in size, respectively.
B: Fragments are shown at ca. 5711 bp for pET16b and 5493 bp for pET22b(+).
All samples shown were digested with the restriction enzymes NdeI and XhoI then run on a 1% agarose gel
with a 1 kb plus DNA Marker.