Part:BBa_K1080001:Design
ChlH
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1928
Illegal BglII site found at 2234
Illegal BglII site found at 3620 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 486
Illegal NgoMIV site found at 2152
Illegal AgeI site found at 1132
Illegal AgeI site found at 2650
Illegal AgeI site found at 2704
Illegal AgeI site found at 2923 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 2254
Illegal BsaI.rc site found at 3049
Illegal SapI.rc site found at 2979
Design Notes
CR-ChlH sequence with His Tag in pET15b. Sequence from Translation start site.
Note 8 PstI sites CTGCAG (785, 1031, 1385, 1652, 2681, 2825, 3458, 3569) No EcoRI or XbaI sites. 1 SpeI (ACTAGT) (4226) site 23 bp after stop codon.
ATGCGGGGTTCTCATCATCATCATCATCATGGTATGGCTAGCATGACTGGTGGAC
AGCAAATGGGTCGGGATCTGTACGACGATGACGATAAGGATCATCCCTTCACCGCGTGCAATGTGGCGACTGGACC
CCGGCCGCCCATGACCACCTTCACCGGTGGCAACAAGGGCCCTGCTAAGCAGCAGGTGTCGCTGGATCTGCGCGAC
GAGGGCGCTGGCATGTTCACCAGCACCAGCCCGGAGATGCGCCGTGTCGTCCCTGACGATGTGAAGGGTCGCGTTA
AGGTGAAGGTTGTGTACGTGGTGCTGGAGGCCCAGTACCAGTCGGCCATCAGCGCTGCGGTGAAGAACATCAACGC
CAAGAACTCCAAGGTGTGCTTCGAGGTGGTGGGCTACCTGCTGGAGGAGCTGCGTGACCAGAAGAACCTCGATATG
CTCAAGGAGGATGTGGCCTCTGCCAACATCTTCATCGGCTCGCTCATCTTCATTGAGGAGCTTGCCGAGAAGATTG
TGGAGGCGGTGAGCCCCCTGCGCGAGAAGCTGGACGCGTGCCTGATCTTCCCGTCCATGCCGGCGGTCATGAAGCT
GAACAAGCTGGGCACGTTTTCGATGGCTCAGCTGGGCCAGTCGAAGTCGGTGTTCTCGGAGTTCATCAAGTCTGCT
CGCAAGAACAACGACAACTTCGAGGAGGGCTTGCTGAAGCTGGTGCGCACCCTGCCTAAGGTGCTGAAGTATCTGC
CCTCGGACAAGGCGCAGGACGCCAAGAACTTCGTGAACAGCCTGCAGTACTGGCTGGGCGGTAACTCGGACAACCT
GGAGAACCTGCTGCTGAACACCGTCAGCAACTACGTGCCCGCTCTGAAGGGCGTGGACTTCAGCGTGGCTGAGCCC
ACCGCCTACCCCGATGTGGGTATCTGGCACCCTCTGGCCTCGGGCATGTACGAGGACCTGAAGGAGTACCTGAACT
GGTACGACACCCGCAAGGACATGGTCTTCGCCAAGGACGCCCCCGTCATTGGCCTGGTGCTGCAGCGCTCGCACCT
GGTGACTGGCGATGAGGGCCACTACAGCGGCGTGGTCGCTGAGCTGGAGAGCCGCGGTGCTAAGGTCATCCCCGTC
TTTGCCGGTGGCCTGGACTTCTCCGCCCCCGTCAAGAAGTTCTTCTACGACCCCCTGGGCTCTGGCCGCACGTTCG
TGGACACCGTTGTGTCGCTGACCGGCTTCGCGCTGGTGGGCGGCCCCGCGCGCCAGGACGCGCCGAAGGCCATTGA
GGCGCTGAAGAACCTGAACGTGCCCTACCTGGTGTCGCTGCCGCTGGTGTTCCAGACCACTGAGGAGTGGCTGGAC
AGCGAGCTGGGCGTGCACCCCGTCCAGGTGGCTCTGCAGGTTGCCCTGCCCGAGCTGGATGGTGCCATGGAGCCCA
TCGTGTTCGCTGGCCGTGACTCGAACACCGGCAAGTCGCACTCGCTGCCCGACCGCATCGCTTCGCTGTGCGCTCG
CGCCGTGAACTGGGCCAACCTGCGCAAGAAGCGCAACGCCGAGAAGAAGCTGGCCGTCACCGTGTTCAGCTTCCCC
CCTGACAAGGGCAACGTCGGCACTGCCGCCTACCTGAACGTGTTCGGCTCCATCTACCGCGTGCTGAAGAACCTGC
AGCGCGAGGGCTACGACGTGGGCGCCCTGCCGCCCTCGGAGGAGGATCTGATCCAGTCGGTGCTGACCCAGAAGGA
GGCCAAGTTCAACTCGACCGACCTGCACATCGCCTACAAGATGAAGGTGGACGAGTACCAGAAGCTGTGCCCTTAC
GCCGAGGCGCTGGAGGAGAACTGGGGCAAGCCCCCCGGCACCCTGAACACCAACGGCCAGGAGCTGCTGGTGTACG
GCCGCCAGTACGGCAACGTCTTCATCGGCGTGCAGCCCACCTTCGGCTACGAGGGCGACCCGATGCGCCTGCTGTT
CTCGAAGTCGGCCAGCCCCCACCACGGCTTCGCCGCCTACTACACCTTCCTGGAGAAGATCTTCAAGGCCGACGCC
GTGCTGCACTTCGGCACCCACGGCTCGCTGGAGTTCATGCCCGGCAAGCAGGTCGGCATGTCGGGTGTGTGCTACC
CCGACTCGCTGATCGGCACCATCCCCAACCTCTACTACTACGCCGCCAACAACCCGTCTGAGGCCACCATCGCCAA
GCGCCGCTCGTACGCCAACACCATTTCGTACCTGACGCCGCCTGCCGAGAACGCCGGCCTGTACAAGGGCCTGAAG
GAGCTGAAGGAGCTGATCAGCTCGTACCAGGGCATGCGTGAGTCTGGCCGCGCCGAGCAGATCTGCGCCACCATCA
TTGAGACCGCCAAGCTGTGCAACCTGGACCGCGACGTGACCCTGCCCGACGCTGACGCCAAGGACCTGACCATGGA
CATGCGCGACAGCGTTGTGGGCCAGGTGTACCGCAAGCTGATGGAGATTGAGTCCCGCCTGCTGCCCTGCGGCCTG
CACGTGGTGGGCTGCCCGCCCACCGCCGAGGAGGCCGTGGCCACCCTGGTCAACATCGCTGAGCTGGACCGCCCGG
ACAACAACCCCCCCATCAAGGGCATGCCCGGCATCCTGGCCCGCGCCATTGGTCGCGACATCGAGTCGATTTACAG
CGGCAACAACAAGGGCGTCCTGGCTGACGTTGACCAGCTGCAGCGCATCACCGAGGCCTCCCGCACCTGCGTGCGC
GAGTTCGTGAAGGACCGCACCGGCCTGAACGGCCGCATCGGCACCAACTGGATCACCAACCTGCTCAAGTTCACCG
GCTTCTACGTGGACCCCTGGGTGCGCGGCCTGCAGAACGGCGAGTTCGCCAGCGCCAACCGCGAGGAGCTGATCAC
CCTGTTCAACTACCTGGAGTTCTGCCTGACCCAGGTGGTCAAGGACAACGAGCTGGGCGCCCTGGTAGAGGCGCTG
AACGGCCAGTACGTCGAGCCCGGCCCCGGCGGTGACCCCATCCGCAACCCCAACGTGCTGCCCACCGGCAAGAACA
TCCACGCCCTGGACCCTCAGTCGATTCCCACTCAGGCCGCGCTGAAGAGCGCCCGCCTGGTGGTGGACCGCCTGCT
GGACCGCGAGCGCGACAACAACGGCGGCAAGTACCCCGAGACCATCGCGCTGGTGCTGTGGGGCACTGACAACATC
AAGACCTACGGCGAGTCGCTGGCCCAGGTCATGATGATGGTCGGTGTCAAGCCCGTGGCCGACGCCCTGGGCCGCG
TGAACAAGCTGGAGGTGATCCCTCTGGAGGAGCTGGGCCGCCCCCGCGTGGACGTGGTTGTCAACTGCTCGGGTGT
GTTCCGCGACCTGTTCGTGAACCAGATGCTGCTGCTGGACCGCGCCATCAAGCTGGCGGCCGAGCAGGACGAGCCC
GATGAGATGAACTTCGTGCGCAAGCACGCCAAGCAGCAGGCGGCGGAGCTGGGCCTGCAGAGCCTGCGCGACGCGG
CCACCCGTGTGTTCTCCAACAGCTCGGGCTCCTACTCGTCCAACGTCAACCTGGCGGTGGAGAACAGCAGCTGGAG
CGACGAGTCGCAGCTGCAGGAGATGTACCTGAAGCGCAAGTCGTACGCCTTCAACTCGGACCGCCCCGGCGCCGGT
GGCGAGATGCAGCGCGACGTGTTCGAGACGGCCATGAAGACCGTGGACGTGACCTTCCAGAACCTGGACTCGTCCG
AGATCTCGCTGACCGATGTGTCGCACTACTTCGACTCCGACCCCACCAAGCTGGTGGCGTCGCTGCGCAACGACGG
CCGCACCCCCAACGCCTACATCGCCGACACCACCACCGCCAACGCGCAGGTCCGCACTCTGGGTGAGACCGTGCGC
CTGGACGCCCGCACCAAGCTGCTCAACCCCAAGTGGTACGAGGGCATGCTTGCCTCGGGCTACGAGGGCGTGCGCG
AGATCCAGAAGCGCATGACCAACACCATGGGCTGGTCGGCCACCTCGGGCATGGTGGACAACTGGGTGTACGACGA
GGCCAACTCGACCTTCATCGAGGATGCGGCCATGGCCGAGCGCCTGATGAACACCAACCCCAACAGCTTCCGCAAG
CTGGTGGCCACCTTCCTGGAGGCCAACGGCCGCGGCTACTGGGACGCCAAGCCCGAGCAGCTGGAGCGCCTGCGCC
AGCTGTACATGGACGTGGAGGACAAGATTGAGGGCGTCGAATAAGCGGCCTCCCCTTCATGGTAGCACTAGTTGGC
GGGTTGTGGTTGGACTAGGCGGCTAGGGTATATACCTAGTAGCGGCGGCTGCGGAGTGGAGGGCTGGCGCCCAGCG
CGAGGGCGTGGCCTTTCCTCCTGGACCCGAGAGCGCTCCGCGAGG
AGACGGCGAGTGAGATAGGCAGCAGCG
10 20 30 40 50 60
MRGSHHHHHH GMASMTGGQQ MGRDLYDDDD KDHPFTACNV ATGPRPPMTT FTGGNKGPAK
70 80 90 100 110 120
QQVSLDLRDE GAGMFTSTSP EMRRVVPDDV KGRVKVKVVY VVLEAQYQSA ISAAVKNINA
130 140 150 160 170 180
KNSKVCFEVV GYLLEELRDQ KNLDMLKEDV ASANIFIGSL IFIEELAEKI VEAVSPLREK
190 200 210 220 230 240
LDACLIFPSM PAVMKLNKLG TFSMAQLGQS KSVFSEFIKS ARKNNDNFEE GLLKLVRTLP
250 260 270 280 290 300
KVLKYLPSDK AQDAKNFVNS LQYWLGGNSD NLENLLLNTV SNYVPALKGV DFSVAEPTAY
310 320 330 340 350 360
PDVGIWHPLA SGMYEDLKEY LNWYDTRKDM VFAKDAPVIG LVLQRSHLVT GDEGHYSGVV
370 380 390 400 410 420
AELESRGAKV IPVFAGGLDF SAPVKKFFYD PLGSGRTFVD TVVSLTGFAL VGGPARQDAP
430 440 450 460 470 480
KAIEALKNLN VPYLVSLPLV FQTTEEWLDS ELGVHPVQVA LQVALPELDG AMEPIVFAGR
490 500 510 520 530 540
DSNTGKSHSL PDRIASLCAR AVNWANLRKK RNAEKKLAVT VFSFPPDKGN VGTAAYLNVF
550 560 570 580 590 600
GSIYRVLKNL QREGYDVGAL PPSEEDLIQS VLTQKEAKFN STDLHIAYKM KVDEYQKLCP
610 620 630 640 650 660
YAEALEENWG KPPGTLNTNG QELLVYGRQY GNVFIGVQPT FGYEGDPMRL LFSKSASPHH
670 680 690 700 710 720
GFAAYYTFLE KIFKADAVLH FGTHGSLEFM PGKQVGMSGV CYPDSLIGTI PNLYYYAANN
730 740 750 760 770 780
PSEATIAKRR SYANTISYLT PPAENAGLYK GLKELKELIS SYQGMRESGR AEQICATIIE
790 800 810 820 830 840
TAKLCNLDRD VTLPDADAKD LTMDMRDSVV GQVYRKLMEI ESRLLPCGLH VVGCPPTAEE
850 860 870 880 890 900
AVATLVNIAE LDRPDNNPPI KGMPGILARA IGRDIESIYS GNNKGVLADV DQLQRITEAS
910 920 930 940 950 960
RTCVREFVKD RTGLNGRIGT NWITNLLKFT GFYVDPWVRG LQNGEFASAN REELITLFNY
970 980 990 1000 1010 1020
LEFCLTQVVK DNELGALVEA LNGQYVEPGP GGDPIRNPNV LPTGKNIHAL DPQSIPTQAA
1030 1040 1050 1060 1070 1080
LKSARLVVDR LLDRERDNNG GKYPETIALV LWGTDNIKTY GESLAQVMMM VGVKPVADAL
1090 1100 1110 1120 1130 1140
GRVNKLEVIP LEELGRPRVD VVVNCSGVFR DLFVNQMLLL DRAIKLAAEQ DEPDEMNFVR
1150 1160 1170 1180 1190 1200
KHAKQQAAEL GLQSLRDAAT RVFSNSSGSY SSNVNLAVEN SSWSDESQLQ EMYLKRKSYA
1210 1220 1230 1240 1250 1260
FNSDRPGAGG EMQRDVFETA MKTVDVTFQN LDSSEISLTD VSHYFDSDPT KLVASLRNDG
1270 1280 1290 1300 1310 1320
RTPNAYIADT TTANAQVRTL GETVRLDART KLLNPKWYEG MLASGYEGVR EIQKRMTNTM
1330 1340 1350 1360 1370 1380
GWSATSGMVD NWVYDEANST FIEDAAMAER LMNTNPNSFR KLVATFLEAN GRGYWDAKPE
1390 1400
QLERLRQLYM DVEDKIEGVE
References and documentation are available.
Please note the modified algorithm for extinction coefficient.
Number of amino acids: 1400
Molecular weight: 154817.9
Theoretical pI: 5.48
Amino acid composition: Ala (A) 116 8.3% Arg (R) 71 5.1% Asn (N) 79 5.6% Asp (D) 86 6.1% Cys (C) 13 0.9% Gln (Q) 48 3.4% Glu (E) 92 6.6% Gly (G) 105 7.5% His (H) 21 1.5% Ile (I) 51 3.6% Leu (L) 142 10.1% Lys (K) 80 5.7% Met (M) 38 2.7% Phe (F) 51 3.6% Pro (P) 71 5.1% Ser (S) 86 6.1% Thr (T) 71 5.1% Trp (W) 14 1.0% Tyr (Y) 50 3.6% Val (V) 115 8.2% 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): 178
Total number of positively charged residues (Arg + Lys): 151
Atomic composition:
Carbon C 6872 Hydrogen H 10826 Nitrogen N 1876 Oxygen O 2091 Sulfur S 51
Formula: C6872H10826N1876O2091S51 Total number of atoms: 21716
Extinction coefficients:
Extinction coefficients are in units of M-1 cm-1, at 280 nm measured in water.
Ext. coefficient 152250 Abs 0.1% (=1 g/l) 0.983, assuming all pairs of Cys residues form cystines
Ext. coefficient 151500
Abs 0.1% (=1 g/l) 0.979, 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 32.82 This classifies the protein as stable.
Aliphatic index: 85.87
Grand average of hydropathicity (GRAVY): -0.292
Source
Chlamydomonas reinhardtii