EG10810

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Author Information

Sharman Tan

Basic Information

  • ID: EG10810
  • Name: pyrG
  • Organism: E. coli
  • Description: Catalyzes the ATP-dependent amination of UTP to CTP with either L-glutamine or ammonia as the source of nitrogen. Regulates intracellular CTP levels through interactions with the four ribonucleotide triphosphates [1]. This protein is involved in step 2 of the subpathway that synthesizes CTP from UDP [2].
  • DNA Length: 1638 base pairs.
  • DNA sequence:

ATG ACT ACG AAT TAC ATC TTT GTT ACT GGT GGC GTG GTC TCA TCC TTA GGG AAA GGT ATT GCA GCA GCT TCG TTA GCC GCA ATT CTT GAA GCG CGC GGG TTG AAT GTA ACA ATT ATG AAA CTG GAC CCT TAC ATC AAC GTG GAT CCA GGC ACT ATG TCG CCA ATT CAA CAC GGC GAA GTT TTT GTG ACG GAA GAT GGC GCC GAA ACA GAC TTA GAC CTG GGC CAC TAT GAA CGC TTC ATC CGC ACT AAG ATG AGT CGT CGT AAC AAC TTC ACC ACT GGG CGT ATT TAC TCG GAC GTG TTA CGT AAA GAA CGT CGT GGG GAC TAT TTG GGT GCA ACT GTT CAG GTG ATC CCA CAT ATC ACA AAC GCG ATC AAA GAG CGT GTA CTG GAG GGG GGG GAA GGC CAT GAC GTA GTT CTT GTT GAG ATT GGA GGA ACA GTG GGT GAT ATC GAG AGT TTA CCC TTT TTA GAA GCT ATT CGC CAA ATG GCT GTT GAA ATC GGA CGC GAG CAT ACG TTA TTT ATG CAT CTG ACA TTG GTC CCC TAC ATG GCC GCC AGT GGT GAG GTG AAA ACT AAA CCG ACT CAG CAT TCA GTA AAG GAA CTT TTA TCC ATC GGC ATT CAG CCT GAC ATC CTT ATT TGC CGC TCG GAT CGC GCA GTC CCA GCT AAC GAG CGC GCT AAA ATT GCA CTT TTC TGT AAC GTG CCT GAG AAG GCG GTC ATT TCG TTA AAG GAC GTT GAT TCG ATC TAC AAA ATT CCA GGG TTA CTG AAA AGT CAA GGA CTT GAC GAT TAC ATC TGC AAG CGC TTT AGC TTG AAT TGC CCA GAA GCC AAC CTG TCG GAA TGG GAG CAA GTT ATT TTC GAG GAG GCG AAC CCG GTG TCC GAA GTC ACG ATT GGA ATG GTA GGA AAG TAT ATC GAA TTG CCA GAC GCA TAC AAG AGC GTT ATC GAG GCC TTA AAG CAT GGG GGC CTT AAA AAC CGT GTT AGT GTA AAC ATC AAA TTG ATC GAT TCA CAA GAC GTG GAA ACT CGC GGG GTT GAA ATT CTG AAG GGT TTG GAT GCT ATC CTG GTT CCC GGG GGG TTC GGA TAC CGC GGG GTA GAG GGA ATG ATT ACC ACC GCA CGT TTT GCC CGT GAG AAC AAT ATC CCG TAC TTG GGT ATT TGT CTT GGA ATG CAA GTT GCA CTG ATC GAT TAC GCC CGC CAC GTC GCA AAC ATG GAG AAC GCT AAT TCT ACG GAA TTT GTC CCG GAC TGT AAA TAT CCC GTA GTT GCT CTG ATC ACT GAG TGG CGT GAC GAG AAT GGA AAT GTG GAA GTT CGT TCT GAG AAG AGC GAC CTG GGC GGT ACC ATG CGC CTG GGC GCC CAG CAA TGC CAA TTG GTT GAC GAT TCC CTT GTT CGT CAA TTG TAC AAT GCC CCG ACT ATC GTG GAA CGT CAC CGT CAC CGT TAT GAA GTA AAT AAC ATG TTG TTA AAG CAG ATC GAA GAC GCC GGA CTT CGC GTA GCT GGA CGT TCT GGT GAT GAT CAA CTG GTG GAA ATC ATC GAG GTG CCT AAC CAC CCG TGG TTC GTA GCG TGC CAA TTT CAC CCC GAG TTT ACT TCG ACA CCC CGT GAT GGT CAT CCC TTA TTC GCT GGA TTC GTG AAA GCA GCT TCG GAG TTT CAA AAG CGC CAA GCG AAA TGA

  • Amino Acid length: 545 amino acids.
  • Amino Acid sequence:

MTTNYIFVTGGVVSSLGKGIAAASLAAILEARGLNVTIMKLDPYINVDPGTMSPIQHGEVFVTEDGAETDLDLGHYERFIRTKMSRRNNFTTGRIYSDVLRKERRGDYLGATVQVIPHITNAIKERVLEGGEGHDVVLVEIGGTVGDIESLPFLEAIRQMAVEIGREHTLFMHLTLVPYMAASGEVKTKPTQHSVKELLSIGIQPDILICRSDRAVPANERAKIALFCNVPEKAVISLKDVDSIYKIPGLLKSQGLDDYICKRFSLNCPEANLSEWEQVIFEEANPVSEVTIGMVGKYIELPDAYKSVIEALKHGGLKNRVSVNIKLIDSQDVETRGVEILKGLDAILVPGGFGYRGVEGMITTARFARENNIPYLGICLGMQVALIDYARHVANMENANSTEFVPDCKYPVVALITEWRDENGNVEVRSEKSDLGGTMRLGAQQCQLVDDSLVRQLYNAPTIVERHRHRYEVNNMLLKQIEDAGLRVAGRSGDDQLVEIIEVPNHPWFVACQFHPEFTSTPRDGHPLFAGFVKAASEFQKRQAK

Function and Homologs

  • Closest homologous proteins: The top (max three) homologous proteins to this protein, as identified by BLAST searches.
    • MULTISPECIES: CTP synthetase [Proteobacteria], Max score: 1125; Query Cover: 100%; E-value: 0.0; Identity: 100%; WP_000210878.1
    • CTP synthase (glutamine hydrolyzing) [Escherichia coli], Max score: 1124, Query Cover: 100%, E-value: 0.0, Identity: 99%, WP_077476003.1
    • CTP synthase (glutamine hydrolyzing) [Shigella sonnei], Max score: 1123, Query Cover: 100%, E-value: 0.0, Identity: 99%, WP_077156148.1
  • Equivalent E. coli / JCVI functional protein: MMSYN1_0129.

Expression

  • Expression Level: High
  • Expression Level Hypothesis: Because CTP is a necessary material in nucleotide synthesis, the expression level for pyrG is most likely high [3]. It is the rate-limiting enzyme for the synthesis of cytosine nucleotides from both the de novo and uridine salvage pathways [4]. Nucleotide synthesis is necessary for cells in the organism to perform DNA and RNA replication, which are necessary processes in order for genes to be expressed. Therefore, it makes sense that pyrG has a high expression level in the organism.
  • Expression Time: Right at the beginning
  • Expression Time Hypothesis: Because CTP is a necessary material in nucleotide synthesis, the expression level for pyrG is most likely high [5]. It is the rate-limiting enzyme for the synthesis of cytosine nucleotides from both the de novo and uridine salvage pathways [6]. Nucleotide synthesis is required for cells to perform RNA and DNA synthesis, which are necessary for the organism to express genes at the beginning of its life cycle. Therefore, it makes sense that the expression time for pyrG is right at the beginning of the organism's life cycle.

Gene Context

  • Other Components: GTPase
  • Possible Dependencies: ATP Synthase
  • Process: ATP-dependent amination of UTP to CTP
    • Inputs: UTP, L-glutamine, ammonia
    • Outputs: CTP
    • Reference: [7]

Construct

  • Synthesis Score: The synthesis score of your construct: 1, 2,3
  • Predicted Translation Rate: Prediction of construct translation rate from the RBS calculator
  • Design Notes and Details: For example, had to use a rare codon to fix folding energy;
  • GenBank File: [8]