Zadatak 1.

-Reverzno komplementarna sekvenca:

CCCCCTCCCTCCCCCACATACTGGCACTTTACTCCTAGACTGTGGGGGACTGTAAAGCCTTCATTCACATCTATCACTGGGGAACACAGCCCGCTTGCTTGACAAACCAAAAAGGATCAA
GTGTGTTTTGTTTTTAAGGCGGAAGATACAAAAAGACACTTGTTGGGTTACATAATTTACAGACAAGCAATTATCATCTAACACCAGTAATTGGTACCGGCTCCTCCCAGCTGGGCTGTT
GCCTTCACAGAGGCGAGTAACTTCCTGTAACAATGCATTATAACAATATTTGGAATGACTATTAAAAAAGAAACAAATGTACAATCAAAGTCCTCAGATGCATTGTAGAACTTTGGGGGC
GTTCGCTCCAACATGTTTCATTTTAAGACTGCTGCTGACACCTTCACCATTCCAGTTTTTTAATCCTGAGTCAAGCGCCAAAAGAAAAAACAAATAAAGCCATGCCAATCTCGTCTTGTTTT
ATGCGCATTTATGGGTTTTGTTTCATCACAGGGGTGTGGGTGTTGGTAACAGTCCGGTTTAGAAGCATTTGCGGTGGACAATGGAGGGTCCGGATTCATCGTACTCCTGCTTGCTGATCC
ACATCTGCTGGAAGGTGGACAGGGAGGCCAGGATAGAGCCTCCAATCCAGACAGAGTACTTGCGCTCAGGTGGGGCAATGATCTTGATTTTCATTGTGCTAGGTGCCAGGGCTGTGAT
CTCCTTCTGCATCCTGTCAGCAATGCCAGGGTACATTGTGGTACCACCAGACAGCACTGTGTTGGCATACAGATCCTTACGGATATCCACATCACACTTCATGATGGAGTTGAAGGTAGT
TTCATGGATACCACAGGACTCCATACCCAAGAAAGATGGCTGGAAGAGGGCCTCGGGGCACCTGAACCTCTCATTGCCAATGGTGATGACCTGACCATCAGGGAGTTCATAGCTCTTCT
CCAGGGAAGAGCTAGAGGCAGCTGTGGCCATCTCCTGCTCGAAATCCAGTGCGACGTAGCACAGCTTCTCCTTGATGTCACGCACAATTTCTCTCTCGGCTGTGGTGGTGAAGCTGTAG
CCTCTCTCTGTCAGGATCTTCATGAGGTAGTCCGTCAGGTCACGGCCAGCCAGATCCAGACGGAGGATGGCATGGGGGAGGGCGTAGCCTTCATAGATGGGCACAGTGTGGGTAACAC
CATCACCAGAGTCCATCACAATACCAGTGGTACGACCAGAGGCATACAGGGACAGCACAGCCTGGATGGCTACATACATGGCTGGGGTGTTGAAGGTCTCAAACATGATCTGTGTCAT
CTTCTCTCTGTTGGCTTTGGGGTTCAGGGGAGCCTCTGTGAGCAGCACAGGGTGCTCCTCAGGGGCTACTCTCAGCTCATTGTAGAAAGTGTGGTGCCAGATCTTCTCCATATCATCCCA
GTTGGTGACAATACCGTGTTCAATGGGGTACTTCAGGGTCAGGATACCTCTTTTGCTCTGGGCTTCATCACCAACGTAGCTGTCTTTCTGGCCCATACCAACCATCACACCCTGATGTCTG
GGGCGACCCACGATAGATGGGAACACAGCACGGGGGGCATCGTCCCCGGCGAAACCGGCCTTGCACATACCGGAGCCATTGTCAACAACGAGCGCAGCAATATCATCATCCATGGCT
GGCTGTGGGAGTAACGCGGTCAGTCAGAGCCGGGGCGGGCGGCGCGAGGCGGCCNNGTTGTTCC

-Komplementarna sekvenca:
CCTTGTTGNNCCGGCGGAGCGCGGCGGGCGGGGCCGAGACTGACTGGCGCAATGAGGGTGTCGGTCGGTACCTACTACTATAACGACGCGAGCAACAACTGTTACCGAGGCCATACA
CGTTCCGGCCAAAGCGGCCCCTGCTACGGGGGGCACGACACAAGGGTAGATAGCACCCAGCGGGGTCTGTAGTCCCACACTACCAACCATACCCGGTCTTTCTGTCGATGCAACCACTA
CTTCGGGTCTCGTTTTCTCCATAGGACTGGGACTTCATGGGGTAACTTGTGCCATAACAGTGGTTGACCCTACTATACCTCTTCTAGACCGTGGTGTGAAAGATGTTACTCGACTCTCATC
GGGGACTCCTCGTGGGACACGACGAGTGTCTCCGAGGGGACTTGGGGTTTCGGTTGTCTCTCTTCTACTGTGTCTAGTACAAACTCTGGAAGTTGTGGGGTCGGTACATACATCGGTAG
GTCCGACACGACAGGGACATACGGAGACCAGCATGGTGACCATAACACTACCTGAGACCACTACCACAATGGGTGTGACACGGGTAGATACTTCCGATGCGGGAGGGGGTACGGTAG
GAGGCAGACCTAGACCGACCGGCACTGGACTGCCTGATGGAGTACTTCTAGGACTGTCTCTCTCCGATGTCGAAGTGGTGGTGTCGGCTCTCTCTTTAACACGCACTGTAGTTCCTCTTC
GACACGATGCAGCGTGACCTAAAGCTCGTCCTCTACCGGTGTCGACGGAGATCGAGAAGGGACCTCTTCTCGATACTTGAGGGACTACCAGTCCAGTAGTGGTAACCGTTACTCTCCAA
GTCCACGGGGCTCCGGGAGAAGGTCGGTAGAAAGAACCCATACCTCAGGACACCATAGGTACTTTGATGGAAGTTGAGGTAGTACTTCACACTACACCTATAGGCATTCCTAGACATAC
GGTTGTGTCACGACAGACCACCATGGTGTTACATGGGACCGTAACGACTGTCCTACGTCTTCCTCTAGTGTCGGGACCGTGGATCGTGTTACTTTTAGTTCTAGTAACGGGGTGGACTCG
CGTTCATGAGACAGACCTAACCTCCGAGATAGGACCGGAGGGACAGGTGGAAGGTCGTCTACACCTAGTCGTTCGTCCTCATGCTACTTAGGCCTGGGAGGTAACAGGTGGCGTTTAC
GAAGATTTGGCCTGACAATGGTTGTGGGTGTGGGGACACTACTTTGTTTTGGGTATTTACGCGTATTTTGTTCTGCTCTAACCGTACCGAAATAAACAAAAAAGAAAACCGCGAACTGA
GTCCTAATTTTTTGACCTTACCACTTCCACAGTCGTCGTCAGAATTTTACTTTGTACAACCTCGCTTGCGGGGGTTTCAAGATGTTACGTAGACTCCTGAAACTAACATGTAAACAAAGAA
AAAATTATCAGTAAGGTTTATAACAATATTACGTAACAATGTCCTTCAATGAGCGGAGACACTTCCGTTGTCGGGTCGACCCTCCTCGGCCATGGTTAATGACCACAATCTACTATTAACG
AACAGACATTTAATACATTGGGTTGTTCACAGAAAAACATAGAAGGCGGAATTTTTGTTTTGTGTGAACTAGGAAAAACCAAACAGTTCGTTCGCCCGACACAAGGGGTCACTATCTAC
ACTTACTTCCGAAATGTCAGGGGGTGTCAGATCCTCATTTCACGGTCATACACCCCCTCCCTCCCCC

1. https://www.genscript.com/sms2/rev_comp.html
2. https://en.vectorbuilder.com/tool/dna-reverse-complement.html
3. https://reverse-complement.com/
 Zadatak 2.
-Nukleotidna sekvenca u proteinsku:
NNXAASRRPPRL*LTALLPQPAMDDDIAALVVDNGSGMCKAGFAGDDAPRAVFPSIVGRP
RHQGVMVGMGQKDSYVGDEAQSKRGILTLKYPIEHGIVTNWDDMEKIWHHTFYNELRVAP
EEHPVLLTEAPLNPKANREKMTQIMFETFNTPAMYVAIQAVLSLYASGRTTGIVMDSGDG
VTHTVPIYEGYALPHAILRLDLAGRDLTDYLMKILTERGYSFTTTAEREIVRDIKEKLCY
VALDFEQEMATAASSSSLEKSYELPDGQVITIGNERFRCPEALFQPSFLGMESCGIHETT
FNSIMKCDVDIRKDLYANTVLSGGTTMYPGIADRMQKEITALAPSTMKIKIIAPPERKYS
VWIGGSILASLSTFQQMWISKQEYDESGPSIVHRKCF*TGLLPTPTPL**NKTHKCA*NK
TRLAWLYLFFLLALDSGLKNWNGEGVSSSLKMKHVGANAPKVLQCI*GL*LYICFFFNSH
SKYCYNALLQEVTRLCEGNSPAGRSRYQLLVLDDNCLSVNYVTQQVSFCIFRLKNKTHLI
LFGLSSKRAVFPSDRCE*RLYSPPQSRSKVPVCGGGRG

-ORF:

Pronađeno 13 ORF

Najbolje izgleda ORF 13:

>lcl|ORF7

MDDDIAALVVDNGSGMCKAGFAGDDAPRAVFPSIVGRPRHQGVMVGMGQK

DSYVGDEAQSKRGILTLKYPIEHGIVTNWDDMEKIWHHTFYNELRVAPEE

HPVLLTEAPLNPKANREKMTQIMFETFNTPAMYVAIQAVLSLYASGRTTG

IVMDSGDGVTHTVPIYEGYALPHAILRLDLAGRDLTDYLMKILTERGYSF

TTTAEREIVRDIKEKLCYVALDFEQEMATAASSSSLEKSYELPDGQVITI

GNERFRCPEALFQPSFLGMESCGIHETTFNSIMKCDVDIRKDLYANTVLS

GGTTMYPGIADRMQKEITALAPSTMKIKIIAPPERKYSVWIGGSILASLS

TFQQMWISKQEYDESGPSIVHRKCF

Zadatak 3.
- Svi enzimi
- Enzyme No. Positions Recognition
name cuts of sites sequence
Acc16I 1 465 tgc/gca
- AccB1I 2 1180 1365 g/gyrcc
- AccBSI 1 1853 gagcgg
- AccI 1 2191 gt/mkac
- AccIII 2 610 1826 t/ccgga
- AcsI 2 35 1458 r/aatty
- AcyI 3 1476 1497 2304 gr/cgyc
- AflIII 2 458 628 a/crygt
- AlwNI 4 127 484 1565 2021 cagnnn/ctg
- ApoI 2 35 1458 r/aatty
- Asp700I 2 37 1155 gaann/nnttc
- AviII 1 465 tgc/gca
- BalI 2 1328 2208 tgg/cca
- BanI 2 1180 1365 g/gyrcc
- BanII 1 585 grgcy/c
- BbiII 3 1476 1497 2304 gr/cgyc
- BbuI 1 2216 gcatg/c
- BcgI 3 354 1543 2084 cgannnnnntgc
- BglI 1 410 gccnnnn/nggc
- BglII 1 415 a/gatct
- BpmI 2 424 1516 ctggag
- BsaAI 1 2220 yac/gtr
- BsaHI 3 1476 1497 2304 gr/cgyc
- BsaMI 5 272 495 529 898 1629 gaatgc
- BsaWI 4 41 610 1644 1826 w/ccggw
- Bse118I 3 1902 1981 2201 r/ccggy
- BseAI 2 610 1826 t/ccgga
- BseRI 1 1448 gaggag
- BsgI 4 601 1389 1724 2076 gtgcag
- BshNI 2 1180 1365 g/gyrcc
- BsiMI 2 610 1826 t/ccgga
- BsmBI 2 1478 1802 cgtctc
- BsmI 5 272 495 529 898 1629 gaatgc
- Bsp13I 2 610 1826 t/ccgga
- Bsp143II 3 860 1104 1872 rgcgc/y
- BspEI 2 610 1826 t/ccgga
- BspHI 1 889 t/catga
- BspLU11I 1 628 a/catgt
- BspMI 1 559 acctgc
- BsrBI 1 1853 gagcgg
- BsrDI 1 1439 gcaatg
- BsrFI 3 1902 1981 2201 r/ccggy
- BssAI 3 1902 1981 2201 r/ccggy
- BstD102I 1 1853 gagcgg
- BstDSI 3 177 1110 1340 c/crygg
- BstH2I 3 860 1104 1872 rgcgc/y
- BstX2I 2 415 1606 r/gatcy
- BstYI 2 415 1606 r/gatcy
- Cfr10I 3 1902 1981 2201 r/ccggy
- CfrI 2 1326 2206 y/ggccr
- DraI 1 92 ttt/aaa
- DsaI 3 177 1110 1340 c/crygg
- EaeI 2 1326 2206 y/ggccr
- Eam1104I 1 1655 ctcttc
- EarI 1 1655 ctcttc
- Eco24I 1 585 grgcy/c
- Eco32I 2 652 2086 gat/atc
- Eco57I 3 490 512 1839 ctgaag
- Eco64I 2 1180 1365 g/gyrcc
- EcoRV 2 652 2086 gat/atc
- Esp3I 2 1478 1802 cgtctc
- FriOI 1 585 grgcy/c
- FspI 1 465 tgc/gca
- GsuI 2 424 1516 ctggag
- HaeII 3 860 1104 1872 rgcgc/y
- Hin1I 3 1476 1497 2304 gr/cgyc
- HincII 2 199 734 gty/rac
- HindII 2 199 734 gty/rac
- HpaI 1 734 gtt/aac
- Hsp92I 3 1476 1497 2304 gr/cgyc
- Kpn2I 2 610 1826 t/ccgga
- Ksp632I 1 1655 ctcttc
- MflI 2 415 1606 r/gatcy
- MluI 1 458 a/cgcgt
- MluNI 2 1328 2208 tgg/cca
- MroI 2 610 1826 t/ccgga
- MscI 2 1328 2208 tgg/cca
- MslI 2 1716 2008 caynn/nnrtg
- Msp17I 3 1476 1497 2304 gr/cgyc
- MspA1I 5 211 506 601 881 1919 cmg/ckg
- Mva1269I 5 272 495 529 898 1629 gaatgc
- NspBII 5 211 506 601 881 1919 cmg/ckg
- NspI 2 632 2216 rcatg/y
- PaeI 1 2216 gcatg/c
- PvuII 2 211 1919 cag/ctg
- RcaI 1 889 t/catga
- SphI 1 2216 gcatg/c
- XcmI 1 965 ccannnnn/nnnntgg
- XhoII 2 415 1606 r/gatcy
- XmnI 2 37 1155 gaann/nnttc
-

- Enzimi koji seku jednom:

- Enzyme No. Positions Recognition
name cuts of sites sequence
Acc16I 1 465 tgc/gca
- AccBSI 1 1853 gagcgg
- AccI 1 2191 gt/mkac
- AviII 1 465 tgc/gca
- BanII 1 585 grgcy/c
- BbuI 1 2216 gcatg/c
- BglI 1 410 gccnnnn/nggc
- BglII 1 415 a/gatct
- BsaAI 1 2220 yac/gtr
- BseRI 1 1448 gaggag
- BspHI 1 889 t/catga
- BspLU11I 1 628 a/catgt
- BspMI 1 559 acctgc
- BsrBI 1 1853 gagcgg
- BsrDI 1 1439 gcaatg
 - BstD102I 1 1853 gagcgg
- DraI 1 92 ttt/aaa
- Eam1104I 1 1655 ctcttc
- EarI 1 1655 ctcttc
- Eco24I 1 585 grgcy/c
- FriOI 1 585 grgcy/c
- FspI 1 465 tgc/gca
- HpaI 1 734 gtt/aac
- Ksp632I 1 1655 ctcttc
- MluI 1 458 a/cgcgt
- PaeI 1 2216 gcatg/c
- RcaI 1 889 t/catga
- SphI 1 2216 gcatg/c
- XcmI 1 965 ccannnnn/nnnntgg

- Enzimi koji seku više puta:

Enzyme No. Positions Recognition

name cuts of sites sequence
AccB1I 2 1180 1365 g/gyrcc
AccIII 2 610 1826 t/ccgga
AcsI 2 35 1458 r/aatty
AcyI 3 1476 1497 2304 gr/cgyc
AflIII 2 458 628 a/crygt
AlwNI 4 127 484 1565 2021 cagnnn/ctg
ApoI 2 35 1458 r/aatty
Asp700I 2 37 1155 gaann/nnttc
BalI 2 1328 2208 tgg/cca
BanI 2 1180 1365 g/gyrcc
BbiII 3 1476 1497 2304 gr/cgyc
BcgI 3 354 1543 2084 cgannnnnntgc
BpmI 2 424 1516 ctggag
BsaHI 3 1476 1497 2304 gr/cgyc
BsaMI 5 272 495 529 898 1629 gaatgc
BsaWI 4 41 610 1644 1826 w/ccggw
Bse118I 3 1902 1981 2201 r/ccggy
BseAI 2 610 1826 t/ccgga
BsgI 4 601 1389 1724 2076 gtgcag
BshNI 2 1180 1365 g/gyrcc
BsiMI 2 610 1826 t/ccgga
BsmBI 2 1478 1802 cgtctc
BsmI 5 272 495 529 898 1629 gaatgc
Bsp13I 2 610 1826 t/ccgga
Bsp143II 3 860 1104 1872 rgcgc/y
BspEI 2 610 1826 t/ccgga
BsrFI 3 1902 1981 2201 r/ccggy
BssAI 3 1902 1981 2201 r/ccggy
BstDSI 3 177 1110 1340 c/crygg
BstH2I 3 860 1104 1872 rgcgc/y
BstX2I 2 415 1606 r/gatcy
BstYI 2 415 1606 r/gatcy
Cfr10I 3 1902 1981 2201 r/ccggy
CfrI 2 1326 2206 y/ggccr
DsaI 3 177 1110 1340 c/crygg
EaeI 2 1326 2206 y/ggccr
Eco32I 2 652 2086 gat/atc
Eco57I 3 490 512 1839 ctgaag
Eco64I 2 1180 1365 g/gyrcc
EcoRV 2 652 2086 gat/atc
Esp3I 2 1478 1802 cgtctc
GsuI 2 424 1516 ctggag
HaeII 3 860 1104 1872 rgcgc/y
Hin1I 3 1476 1497 2304 gr/cgyc
HincII 2 199 734 gty/rac
HindII 2 199 734 gty/rac
Hsp92I 3 1476 1497 2304 gr/cgyc
Kpn2I 2 610 1826 t/ccgga
MflI 2 415 1606 r/gatcy
MluNI 2 1328 2208 tgg/cca
MroI 2 610 1826 t/ccgga
MscI 2 1328 2208 tgg/cca
MslI 2 1716 2008 caynn/nnrtg
Msp17I 3 1476 1497 2304 gr/cgyc
MspA1I 5 211 506 601 881 1919 cmg/ckg
Mva1269I 5 272 495 529 898 1629 gaatgc
NspBII 5 211 506 601 881 1919 cmg/ckg
NspI 2 632 2216 rcatg/y
PvuII 2 211 1919 cag/ctg
XhoII 2 415 1606 r/gatcy
XmnI 2 37 1155 gaann/nnttc

Zadatak 4.
- Najduži ORF:
MVPCFSHPYTNILLPYSSGCLRPTKAESLLQLPCASDPVVLLKYFSHLFI

PFTPIAPTSHCHQAVTMQYVSY

- MW I pI iznose: 7.72 / 8072.49

Zadatak 5.
Zadatak 6.
- Sooksa-Nguan, 2009
Sooksa-Nguan, T., Yakubov, B., Kozlovskyy, V. I., Barkume, C. M., Howe, K. J.,
Thannhauser, T. W., ... & Vatamaniuk, O. K. (2009). Drosophila ABC transporter, DmHMT-1,
confers tolerance to cadmium: DmHMT-1 and its yeast homolog, SpHMT-1, are not essential
for vacuolar phytochelatin sequestration. Journal of Biological Chemistry, 284(1), 354-362.

- Hull, 2014
Hull, J. J., Chaney, K., Geib, S. M., Fabrick, J. A., Brent, C. S., Walsh, D., & Lavine, L. C.
(2014). Transcriptome-based identification of ABC transporters in the western tarnished plant
bug Lygus hesperus. PLoS One, 9(11), e113046.

- Mireji, 2010
 Mireji, P. O., Keating, J., Hassanali, A., Impoinvil, D. E., Mbogo, C. M., Muturi, M. N., ... &
Beier, J. C. (2010). Expression of metallothionein and α-tubulin in heavy metal-tolerant
Anopheles gambiae sensu stricto (Diptera: Culicidae). Ecotoxicology and environmental
safety, 73(1), 46-50.

- Mattingly, 2001
Mattingly, K. S., Beaty, B. J., Mackie, R. S., McGaw, M., Carlson, J. O., & Rayms-Keller, A.
(2001). Molecular cloning and characterization of a metal responsive Chironomus tentans
alpha-tubulin cDNA. Aquatic Toxicology, 54(3-4), 249-260.

- Merritt, 2017
Merritt, T. J., & Bewick, A. J. (2017). Genetic diversity in insect metal tolerance. Frontiers in
genetics, 8, 172.

- Salnikow, 2008
Salnikow, K., & Zhitkovich, A. (2008). Genetic and epigenetic mechanisms in metal
carcinogenesis and cocarcinogenesis: nickel, arsenic, and chromium. Chemical research in
toxicology, 21(1), 28-44.