Part:BBa_K1080002:Design
ChlD
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 1334
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 2039
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 614
Illegal NgoMIV site found at 1433
Illegal NgoMIV site found at 1975 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
Incorporated sequence overlap for Gibson assembly and no GC rich region or restriction site in sequence
ChlD 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
Note: No XbaI, EcoRI, PstI or SpeI sites
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
CTGCGCGCCATGAAGGTGTCTGAGGAGGACTCCAAGGGCTTCGATGCGGATGTGTCGACCCGCCTGGCCCGCTCGTACCCTCTGGCGGCCGTGGTGGGCCAGGACAACATCAAGCAGGCGCTGCTGCTGGGCGCCGTGGACACCGGGCTGGGCGGCATCGCCATCGCCGGTCGCCGCGGTACCGCCAAGTCCATCATGGCTCGCGGCCTGCACGCTCTGCTGCCGCCCATTGAGGTGGTGGAGGGCAGCATCTGCAACGCCGACCCCGAGGACCCCCGCTCCTGGGAGGCTGGCCTGGCTGAGAAGTATGCGGGCGGCCCTGTGAAGACCAAGATGCGCTCGGCGCCGTTTGTGCAGATCCCTCTGGGTGTGACTGAGGACCGCTTGGTGGGCACTGTGGACATTGAGGCGTCCATGAAGGAGGGCAAGACTGTGTTCCAGCCCGGCCTGCTGGCTGAGGCGCACCGCGGCATCCTGTACGTGGACGAGATCAACCTGCTGGATGACGGCATTGCCAACCTGCTGCTGTCCATCCTGTCGGACGGAGTCAACGTGGTGGAGCGCGAGGGCATCTCCATCAGCCACCCCTGCCGGCCGCTGCTGATTGCCACCTACAACCCCGAGGAGGGCCCTCTGCGTGAGCACCTGCTGGACCGCATCGCCATTGGCCTCAGCGCCGACGTCCCCAGCACCAGCGACGAGCGCGTCAAGGCCATTGACGCAGCCATCCGCTTCCAGGACAAGCCGCAGGACACTATTGACGACACCGCGGAGCTCACCGACGCCCTGCGCACCTCGGTCATCCTGGCTCGCGAGTACCTGAAGGACGTGACCATCGCGCCGGAGCAGGTGACCTACATTGTGGAGGAGGCGCGCCGCGGCGGAGTCCAGGGGCACCGCGCGGAGCTGTACGCGGTCAAGTGTGCCAAGGCGTGTGCGGCTCTGGAGGGCCGTGAGCGTGTGAACAAGGATGACCTGCGCCAGGCCGTGCAGCTGGTCATCCTGCCGCGCGCCACCATCCTGGACCAGCCCCCGCCCGAGCAGGAGCAGCCCCCGCCGCCGCCCCCGCCCCCTCCCCCGCCGCCGCCGCAGGACCAAATGGAGGACGAGGACCAGGAGGAGAAGGAGGACGAGAAGGAGGAGGAGGAGAAGGAGAACGAGGACCAGGACGAGCCCGAGATCCCTCAGGAGTTCATGTTTGAGTCCGAGGGCGTCATCATGGACCCCTCCATCCTCATGTTCGCGCAGCAGCAGCAGCGCGCGCAGGGCCGCTCCGGCCGCGCCAAGACGCTCATCTTCAGCGACGACCGCGGCCGCTACATCAAGCCCATGCTGCCCAAGGGTGACAAGGTCAAGCGCCTGGCAGTGGACGCCACGCTTCGCGCCGCCGCGCCCTACCAGAAGATTCGCCGGCAGCAGGCCATCAGCGAGGGCAAGGTGCAGCGCAAGGTGTACGTGGACAAGCCAGACATGCGCTCCAAGAAGCTGGCCCGCAAGGCCGGTGCGCTGGTGATTTTTGTTGTGGACGCGTCCGGCTCCATGGCTCTGAACCGCATGAGCGCCGCCAAGGGCGCCTGCATGCGCCTGCTGGCTGAGTCGTACACCAGCCGCGACCAGGTGTGCCTCATCCCCTTCTACGGCGACAAGGCCGAGGTGCTGCTGCCGCCCTCCAAGTCCATCGCCATGGCCCGCCGCCGCCTGGACTCGCTGCCCTGCGGCGGCGGCTCGCCCCTTGCGCACGGCCTGTCCACGGCGGTACGTGTGGGCATGCAGGCCAGCCAGGCGGGCGAGGTGGGCCGCGTCATGATGGTGCTCATCACGGACGGCCGCGCCAACGTCAGCCTGGCCAAGTCCAACGAGGACCCCGAGGCGCTCAAGCCCGACGCGCCCAAGCCCACCGCCGACTCGCTGAAGGACGAGGTGCGCGACATGGCCAAGAAGGCCGCGTCCGCCGGCATCAACGTGCTTGTCATTGACACGGAGAACAAGTTCGTGAGCACCGGCTTTGCGGAGGAGATCTCCAAGGCAGCGCAGGGCAAGTACTACTACCTGCCCAACGCCAGCGACGCCGCCATCGCGGCGGCCGCGTCCGGCGCCATGGCCGCGGCCAAGGGCGGCTACTAGGTGCCGAGTGACTGAGGTGGCAAGGTGCAGTGGCGGCGGAGGCAGTTGTGCTGGGGTGGCAAGGCGGACAGGCGAAGCTGGTGGGTTGCGACGAGGAGGAGGTGCACGTGCACGCGTAACATAAGAAGAACAGTGGGAGGACAGGTAGCGTGACTTGACTGGGACGAGGAGCGTACTGATGTGTGGCGTGTGTTGGTATGTGAGCGTTACCCCTCC </b></span>
Protein sequence:
10 20 30 40 50 60
MRGSHHHHHH GMASMTGGQQ MGRDLYDDDD KDHPFTLRAM KVSEEDSKGF DADVSTRLAR
70 80 90 100 110 120
SYPLAAVVGQ DNIKQALLLG AVDTGLGGIA IAGRRGTAKS IMARGLHALL PPIEVVEGSI
130 140 150 160 170 180
CNADPEDPRS WEAGLAEKYA GGPVKTKMRS APFVQIPLGV TEDRLVGTVD IEASMKEGKT
190 200 210 220 230 240
VFQPGLLAEA HRGILYVDEI NLLDDGIANL LLSILSDGVN VVEREGISIS HPCRPLLIAT
250 260 270 280 290 300
YNPEEGPLRE HLLDRIAIGL SADVPSTSDE RVKAIDAAIR FQDKPQDTID DTAELTDALR
310 320 330 340 350 360
TSVILAREYL KDVTIAPEQV TYIVEEARRG GVQGHRAELY AVKCAKACAA LEGRERVNKD
370 380 390 400 410 420
DLRQAVQLVI LPRATILDQP PPEQEQPPPP PPPPPPPPPQ DQMEDEDQEE KEDEKEEEEK
430 440 450 460 470 480
ENEDQDEPEI PQEFMFESEG VIMDPSILMF AQQQQRAQGR SGRAKTLIFS DDRGRYIKPM
490 500 510 520 530 540
LPKGDKVKRL AVDATLRAAA PYQKIRRQQA ISEGKVQRKV YVDKPDMRSK KLARKAGALV
550 560 570 580 590 600
IFVVDASGSM ALNRMSAAKG ACMRLLAESY TSRDQVCLIP FYGDKAEVLL PPSKSIAMAR
610 620 630 640 650 660
RRLDSLPCGG GSPLAHGLST AVRVGMQASQ AGEVGRVMMV LITDGRANVS LAKSNEDPEA
670 680 690 700 710 720
LKPDAPKPTA DSLKDEVRDM AKKAASAGIN VLVIDTENKF VSTGFAEEIS KAAQGKYYYL
730 740
PNASDAAIAA AASGAMAAAK GGY
References and documentation are available.
Please note the modified algorithm for extinction coefficient.
Number of amino acids: 743
Molecular weight: 80527.5
Theoretical pI: 5.37
Amino acid composition: Ala (A) 89 12.0% Arg (R) 49 6.6% Asn (N) 14 1.9% Asp (D) 57 7.7% Cys (C) 7 0.9% Gln (Q) 33 4.4% Glu (E) 55 7.4% Gly (G) 56 7.5% His (H) 13 1.7% Ile (I) 40 5.4% Leu (L) 62 8.3% Lys (K) 44 5.9% Met (M) 23 3.1% Phe (F) 13 1.7% Pro (P) 51 6.9% Ser (S) 44 5.9% Thr (T) 26 3.5% Trp (W) 1 0.1% Tyr (Y) 17 2.3% Val (V) 49 6.6% 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): 112
Total number of positively charged residues (Arg + Lys): 93
Atomic composition:
Carbon C 3504 Hydrogen H 5684 Nitrogen N 1008 Oxygen O 1102 Sulfur S 30
Formula: C3504H5684N1008O1102S30 Total number of atoms: 11328
Extinction coefficients:
Extinction coefficients are in units of M-1 cm-1, at 280 nm measured in water.
Ext. coefficient 31205 Abs 0.1% (=1 g/l) 0.388, assuming all pairs of Cys residues form cystines
Ext. coefficient 30830
Abs 0.1% (=1 g/l) 0.383, 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 44.71 This classifies the protein as unstable.
Aliphatic index: 84.64
Grand average of hydropathicity (GRAVY): -0.392
Source
Chlamydomonas reinhardtii