Primers
Table1 Creatinine degradation
| Primers |
Primer Sequence(5' to 3') |
| crnA |
F:GGGAAAGGTACCATGAGCAAGAGTGTTTTTGTAGGTGAGC |
| R:CATGGTATATCTCCTGGATCCTTAGGTGGGCGGGAACTCCTC |
| creJ |
F:AAGCTAGCAGGAGATATACCATGACTACCGCCAACATCGCCA |
| R:GGTATATCTCCTGCTAGCTTTACGAATCGGTTCCTGCAGGC |
| crnA-creJ-racE over-lap |
F:TTAAAGAGGAGAAAGGTACCATGAGCAAGAGTGTTTTTGTAGGTGAGC |
| R:CCTCTAGCACGCGTACCATGTTACGCGTCGATGATGTTGTTCTCCG |
| pCS backbone |
F：CATGGTACGCGTGCTAGAGGC |
| R:GGTATATCTCCTGCTAGCTTTACGAATCGGTTCCTGCAGGC |
Table2 Urea degradation
| Primers |
Primer Sequence(5' to 3') |
| ureABC |
F:TTAAAGAGGAGAAAGGTACCATGGAATTAACCCCCAGAGAAAAAGATAAATTATTGC |
| R:GCACGCGTACCATGGGATCCTTAGAATAAAAAGTAACGTTGTGCCATCGGTAAC |
| ureEFGD |
F:TTAAAGAGGAGAAAGGTACCATGGAATTAACCCCCAGAGAAAAAGATAAATTATTGC |
| R:GCACGCGTACCATGGGATCCTTAGAATAAAAAGTAACGTTGTGCCATCGGTAAC |
| lac-ureABC |
F:TACTAGCGCAGCTTAATTAAAATTGTGAGCGGATAACAATTGACATTGT |
| R:GCACGCGTACCATGGGATCCTTAGAATAAAAAGTAACGTTGTGCCATCGGTAAC |
| lac-ureEFGD |
F:TACTAGCGCAGCTTAATTAAAATTGTGAGCGGATAACAATTGACATTGT |
| R:GATATAGTTCCTCCTTTCAGACAACAGATAAAACGAAAGGCCCAGT |
| lac-ureABC-lac-ureABC Overlap |
F:TACTAGCGCAGCTTAATTAAAATTGTGAGCGGATAACAATTGACATTGT |
| R:GATATAGTTCCTCCTTTCAGACAACAGATAAAACGAAAGGCCCAGT |
Table3 Polyglutamic acid production
| Primers |
Primer Sequence(5' to 3') |
| racE |
F:AAGGATCCAGGAGATATACCATGGAACAGCCGATTGGTGTTATTGATAG |
| R:GGTATATCTCCTGCTAGCTTTACGAATCGGTTCCTGCAGGC |
| pgsBCAE |
F:GGGAAAGGTACCATGTGGTTACTCATTATAGCCTGTGCT |
| R:CAATAAATAAGGATCCCATGGTACGCGTGCTAGAGGCAT |
Table4 FULL ACCESS
| Primers |
Primer Sequence(5' to 3') |
| lac-ureABC-T1 |
F:CTTAATTAACCTAGGCTGCTGCCACCGCTGAATTGTGAGCGGATAACAATTGACATTGT |
| R:GGGTTATGCTAGTTATTGCTACAACAGATAAAACGAAAGGCCCAGT |
| plpp1.2-ureEFGD |
F:GCCTAGGTTAATTAAGCTGCGCTAGTAGACGAGTCCATCAAAAAAATATTGACAACATAAAAAACTTTGTGTGAA |
| R:CTCGAGTCTGGTAAAGAAACCGCTGCTGCGAAATTTGAACGCCAGCACATATTTGTCCTACTCAGGAGAGCGTTC |
| lac-pgsBCAE-T1 |
F:CTTAATTAACCTAGGCTGCTGCCACCGCTGAATTGTGAGCGGATAACAATTGACATTGT |
| R:GGGTTATGCTAGTTATTGCTACAACAGATAAAACGAAAGGCCCAGT |
| pETlac backbone |
F:GGGTTATGCTAGTTATTGCTACAACAGATAAAACGAAAGGCCCAGT |
| R:GGGAAAGGATCCTTATTTAGATTTTAGTTTATCGCTATGATCAACATCAAACGTC |
Table5 Kill switch
| Primers |
Primer Sequence(5' to 3') |
| TtgR |
F:AATTCATTAAAGAGGAGAAAGGTACCATGGTCCGTCGAACCAAAGAAGA |
| R:TTGGTATATCTCCTAAGCTTTCATTTGCGCAGAGCCGG |
| asrG(SR4) |
F:AAGCTTAGGAGATATACCAATGATACAATTATATAATTATTTTTTGCATTTTGTTTAATTAAATGCAT |
| R:TTTTATTTGATGCCTCTAGAAAATAGACCTTCCCTTGAGTTTGGACAC |
| Pttg |
F:GGATTTTGGTCATGACTAGTCAGCAGTATTTACAAACAACCATGAATGT |
| R:GGTACCTTTCTCCTCTTTAATGAATTCAAATAGCT |
| bsrG |
F:AATTCATTAAAGAGGAGAAAGGTACCATGGTCCGTCGAACCAAAGAAGA |
| R:GCACGCGTACCATGGGATCCCTATTTTTTCTTTTGGCTTGACGTTACAATC |
| pZe12-glnAp2 backbone |
F:GGATCCCATGGTACGCGTGC |
| R:TACATTCATGGTTGTTTGTAAATACTGCTGCTCGAGGTGAAGACGAAAGGGC |
Methods
Medium formulation
Table6 LB culture and substrate formulation
| Component |
Content(g/L) |
| yeast extraction |
5 |
| Trptone |
10 |
| NaCl |
10 |
Table7 M9 culture and substrate formulation
| Component |
Content(g/L) |
| Na2HPO4 |
6.78 |
| KH2PO4 |
3 |
| NaCl |
1 |
| NH4Cl |
1 |
| yeast extraction |
5 |
| Mops |
2 |
Add glc before use.
If there is no nitrogen source, remove NH4Cl and yeast extraction