Part:BBa_K1080005:Design
CTH1
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 305
Illegal BglII site found at 413 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 103
Illegal BsaI.rc site found at 259
Illegal BsaI.rc site found at 880
Design Notes
Incorporated sequence overlap for Gibson assembly and no GC rich region or restriction site in sequence
CTH1 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
One PstI site at 740. No EcoRi, SpeI or XbaI
DNA-Sequence: ATGCGGGGTTCTCATCATCATCATCATCATGGTATGGCTAGCATGACTGGT
GGACAGCAAATGGGTCGGGATCTGTACGACGATGACGATAAGGATCATCCC TTCACC<b>
CAGGCCAGCGGGAATGAGCAGCAGTCTCGCCGGGTCCGTGTGGCTGCTACCGCTGCTCCCCAGGAGGTTGAGGGCTTCAAGGTTATGCGCGACGGCATCAAGGTTGCTTCCGATGAGACCCTGCTCACTCCTCGCTTCTACACCACCGACTTCGATGAGATGGAGCGCCTGTTCAGCCTGGAGCTGAACAAGAACATGGACATGGAGGAGTTCGAGGCGATGCTGAACGAGTTCAAGCTGGACTACAACCAGCGCCACTTCGTCCGCAACGAGACCTTCAAGGAGGCCGCGGAGAAGATTCAGGGCCCCACCCGCAAGATTTTCATTGAGTTCCTGGAGCGCTCGTGCACTGCTGAGTTCTCGGGCTTCCTGCTCTACAAGGAGCTGGGTCGCCGCCTGAAGGCCACGAACCCTGTGGTCGCGGAGATCTTCACCCTGATGTCCCGTGACGAGGCCCGCCACGCCGGCTTCCTGAACAAGGCCATGTCCGACTTCAACCTGGCTCTGGACCTGGGTTTCCTGACCAAGAACCGCAAGTACACCTTCTTCAAGCCCAAGTTCATCTTCTACGCCACTTACCTGTCGGAGAAGATTGGCTACTGGCGCTACATTTCTATCTACCGCCACCTGCAGCGCAACCCCGACAACCAGCTGTACCCGCTGTTCGAGTACTTCGAGAACTGGTGCCAGGACGAGAACCGCCACGGTGACTTCTTCACCGCCGTGCTGAAGGCGCGGCCGGAGATGGTCAACGACTGGGCTGCCAAGCTGTGGTCGCGCTTTTTCTGCCTGTCGGTGTACATCACCATGTACCTGAACGACCACCAGCGCGACGCCTTCTACAGCTCTCTGGGCCTGAACACCACCCAGTTCAACCAGCACGTGATCATCGAGACCAACAAGTCTACTGAGCGTATCTTCCCCGCTGTGCCGGATGTGGAGAACCCCGAGTTCTTCCGCCGCATGGACCTGCTGGTCAAGTACAACGCCCAGCTGGTGAACATCGGCAGCATGAACCTGCCCAGCCCCATCAAGGCGATCATGAAGGCGCCCATTCTGGAGCGCATGGTCGCTGAGGTGTTCCAGGTGTTCATCATGACCCCTAAGGAGTCGGGCTCGTACGACCTGGATGCCAACAAGACCGCTCTGGTCTACTAAGCGGCTGGCGTACTAGCCTGCGGGAGCTGGGCGGCTGGATGTCGGAGTTTTGAGAGTGCTTTGGAGCTCGGCGGCGAGCAGCAGCTGTGTGTGGCAGCGTGTGTGTGCAGCAGCAAGTGTAGGTATACAGCCAGTGGCGCTGCTCGCTGGCGGCGCGGTGTCGTGTTGGGCGCCGTCCGACAGCGCCGCGGCGGCCCGCTGCCGTAGTCGACGCGGGTTGGTCACTCATCCAGGAGACGACCTCTCGAGTAGCTGATAGCTTGAGATGAGGGTTCTCTATGCGC
<b>PROTEIN Sequence:
MRGSHHHHHHGMASMTGGQQMGRDLYDDDDKDHPFTQASGNEQQSRRVRVAATAAPQEVE GFKVMRDGIKVASDETLLTPRFYTTDFDEMERLFSLELNKNMDMEEFEAMLNEFKLDYNQ RHFVRNETFKEAAEKIQGPTRKIFIEFLERSCTAEFSGFLLYKELGRRLKATNPVVAEIF TLMSRDEARHAGFLNKAMSDFNLALDLGFLTKNRKYTFFKPKFIFYATYLSEKIGYWRYI SIYRHLQRNPDNQLYPLFEYFENWCQDENRHGDFFTAVLKARPEMVNDWAAKLWSRFFCL SVYITMYLNDHQRDAFYSSLGLNTTQFNQHVIIETNKSTERIFPAVPDVENPEFFRRMDL LVKYNAQLVNIGSMNLPSPIKAIMKAPILERMVAEVFQVFIMTPKESGSYDLDANKTALV Y
Number of amino acids: 421
Molecular weight: 49365.1
Theoretical pI: 6.25
Amino acid composition: Ala (A) 30 7.1% Arg (R) 28 6.7% Asn (N) 24 5.7% Asp (D) 25 5.9% Cys (C) 3 0.7% Gln (Q) 16 3.8% Glu (E) 32 7.6% Gly (G) 18 4.3% His (H) 13 3.1% Ile (I) 18 4.3% Leu (L) 36 8.6% Lys (K) 24 5.7% Met (M) 18 4.3% Phe (F) 34 8.1% Pro (P) 16 3.8% Ser (S) 21 5.0% Thr (T) 23 5.5% Trp (W) 4 1.0% Tyr (Y) 18 4.3% Val (V) 20 4.8% Pyl (O) 0 0.0% Sec (U) 0 0.0% (B) 0 0.0% (Z) 0 0.0% (X) 0 0.0%
Total number of negatively charged residues (Asp + Glu): 57
Total number of positively charged residues (Arg + Lys): 52
Atomic composition:
Carbon C 2222 Hydrogen H 3379 Nitrogen N 599 Oxygen O 638 Sulfur S 21
Formula: C2222H3379N599O638S21 Total number of atoms: 6859
Extinction coefficients:
Extinction coefficients are in units of M-1 cm-1, at 280 nm measured in water.
Ext. coefficient 48945 Abs 0.1% (=1 g/l) 0.991, assuming all pairs of Cys residues form cystines
Ext. coefficient 48820
Abs 0.1% (=1 g/l) 0.989, assuming all Cys residues are reduced
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 37.56 This classifies the protein as stable.
Aliphatic index: 70.93
Grand average of hydropathicity (GRAVY): -0.477
Source
Chlamydomonas reinhardtii