Part:BBa_K1080000:Design
ChlI1
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 231
Illegal BglII site found at 887
Illegal BglII site found at 1082
Illegal BamHI site found at 279 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
Incorporated sequence overlap for Gibson assembly and no GC rich region or restriction site in sequence
ChlI1 Clone: DNA sequence from translation start site: Regions in BOLD are the sequence of the leader region in the pET100 plasmid. Translated the DNA sequence into a protein sequence using "Translate" at http://au.expasy.org/tools Then used the translated protein sequence to analyse the protein using "ProtParam" at http://au.expasy.org/tools
No Biobrick restriction sites (EcoRI, XbaI, SpeI or PstI). Note This won’t work with the highlighted tag in the magnesium chelatase reaction.
ATG CGG GGT TCT CAT CAT CAT CAT CAT CAT GGT ATG GCT AGC ATG ACT GGT GGA
CAG CAA ATG GGT CGG GAT CTG TAC GAC GAT GAC GAT AAG GAT CAT CCC TTC ACC
GCTGCCGCTACTGAG
GTCAAGGCTGCTGAGGGCCGCACTGAGAAGGAGCTGGGCCAGGCCCGCCCCATCTTCCCC
TTCACCGCCATCGTGGGCCAGGATGAGATGAAGCTGGCGCTGATTCTGAACGTGATCGAC
CCCAAGATCGGTGGTGTCATGATCATGGGCGACCGTGGCACTGGCAAGTCCACCACCATT
CGTGCCCTGGCGGATCTGCTGCCCGAGATGCAGGTGGTTGCCAACGACCCCTTTAACTCG
GACCCCACCGACCCCGAGCTGATGAGCGAGGAGGTGCGCAACCGCGTCAAGGCCGGCGAG
CAGCTGCCCGTGTCTTCCAAGAAGATTCCCATGGTGGACCTGCCCCTGGGCGCCACTGAG
GACCGCGTGTGCGGCACCATCGACATCGAGAAGGCGCTGACCGAGGGTGTCAAGGCGTTC
GAGCCCGGCCTGCTGGCCAAGGCCAACCGCGGCATCCTGTACGTGGATGAGGTCAACCTG
CTGGACGACCACCTGGTCGATGTGCTGCTGGACTCGGCCGCCTCCGGCTGGAACACCGTG
GAGCGCGAGGGTATCTCCATCAGCCACCCCGCCCGCTTCATCCTGGTCGGCTCGGGCAAC
CCCGAGGAGGGTGAGCTGCGCCCCCAGCTGCTGGATCGCTTCGGCATGCACGCCCAGATC
GGCACCGTCAAGGACCCCCGCCTGCGTGTGCAGATCGTGTCGCAGCGCTCGACCTTCGAC
GAGAACCCCGCCGCCTTCCGCAAGGACTACGAGGCCGGCCAGATGGCGCTGACCCAGCGC
ATCGTGGACGCGCGCAAGCTGCTGAAGCAGGGCGAGGTCAACTACGACTTCCGCGTCAAG
ATCAGCCAGATCTGCTCGGACCTGAACGTGGACGGCATCCGCGGCGACATCGTGACCAAC
CGCGCCGCCAAGGCCCTGGCCGCCTTCGAGGGCCGCACCGAGGTGACCCCCGAGGACATC
TACCGTGTCATTCCCCTGTGCCTGCGCCACCGCCTCCGGAAAGACCCCCTGGCTGAGATC
GACGACGGTGACCGCGTGCGTGAGATCTTCAAGCAGGTGTTCGGCATGGAGTAAGCGGCA
Amino Acid sequence:
MRGSHHHHHHGMASMTGGQQMGRDLYDDDDKDHPFTAAATEVKAAEGRTEKELGQARPIF PFTAIVGQDEMKLALILNVIDPKIGGVMIMGDRGTGKSTTIRALADLLPEMQVVANDPFN SDPTDPELMSEEVRNRVKAGEQLPVSSKKIPMVDLPLGATEDRVCGTIDIEKALTEGVKA FEPGLLAKANRGILYVDEVNLLDDHLVDVLLDSAASGWNTVEREGISISHPARFILVGSG NPEEGELRPQLLDRFGMHAQIGTVKDPRLRVQIVSQRSTFDENPAAFRKDYEAGQMALTQ RIVDARKLLKQGEVNYDFRVKISQICSDLNVDGIRGDIVTNRAAKALAAFEGRTEVTPED IYRVIPLCLRHRLRKDPLAEIDDGDRVREIFKQVFGME
Number of amino acids: 398
Molecular weight: 44018.0
Theoretical pI: 5.35
Amino acid composition:
Ala (A) 33 8.3% Arg (R) 30 7.5% Asn (N) 12 3.0% Asp (D) 35 8.8% Cys (C) 3 0.8% Gln (Q) 15 3.8% Glu (E) 29 7.3% Gly (G) 33 8.3% His (H) 11 2.8% Ile (I) 25 6.3% Leu (L) 35 8.8% Lys (K) 20 5.0% Met (M) 13 3.3% Phe (F) 13 3.3% Pro (P) 20 5.0% Ser (S) 16 4.0% Thr (T) 19 4.8% Trp (W) 1 0.3% Tyr (Y) 5 1.3% Val (V) 30 7.5%
Asx (B) 0 0.0% Glx (Z) 0 0.0% Xaa (X) 0 0.0%
Total number of negatively charged residues (Asp + Glu): 64 Total number of positively charged residues (Arg + Lys): 50
Atomic composition:
Carbon C 1920 Hydrogen H 3100 Nitrogen N 558 Oxygen O 594 Sulfur S 16
Formula: C1920H3100N558O594S16 Total number of atoms: 6188
Extinction coefficients:
Extinction coefficients are in units of M-1 cm-1, at 280 nm.
Ext. coefficient 13075 Abs 0.1% (=1 g/l) 0.297, assuming ALL Cys residues appear as half cystines
Ext. coefficient 12950 Abs 0.1% (=1 g/l) 0.294, assuming NO Cys residues appear as half cystines
Estimated half-life:
The N-terminal of the sequence considered is M (Met).
The estimated half-life is:
30 hours (mammalian reticulocytes, in vitro). >20 hours (yeast, in vivo). >10 hours (Escherichia coli, in vivo).
Instability index:
The instability index (II) is computed to be 28.58 This classifies the protein as stable.
Aliphatic index: 88.94
Grand average of hydropathicity (GRAVY): -0.367
Source
Chlamydomonas reinhardtii