Silly Lab Tech!
A Plasmid Activity
New England Bio Labs- NEB Cutter - website for predicting restriction maps
Your limited results for some digests
Background:
You have been carrying out experiments on various plasmids and after weeks of work constructing plasmids
you are horrified to find your lab assistant has left your precious plasmids tubes unlabeled! Silly lab tech!
You have no idea which plasmid is found within three tubes! You will need to reidentify each plasmid!
This means completing a restriction digest on each plasmid to confirm the identity of each tube.
You have been working on three different plasmids: pGlo, pAmp, pKan but again you are not sure which tube contains
the right plasmid. Below are the DNA sequences for each plasmid.
You will have the following restriction enzymes to work with HindIII, BamHI, EcoRI, AseI, HpaI
You direct your silly lab tech to digest a small amount of the plasmid with each enzyme. But your lab tech
strikes again and only completes some of the digests and runs out of plasmid so you cannot run more digests!
So you will need to identify the tubes by comparing your expected length of fragments from your plasmid maps
to what is shown in your lab techs limited gel results linked above.
Directions:
1. You will need to use a website to construct a circular plasmid map for each plasmid below and map the restriction
sites using the above restriction enzymeson each plasmid. This will be turned in as part of your grade. (3 points each)
Some guidance about NEBcutter (link above): After cutting and pasting your sequence into the text box on
NEBCutter, click "circular", and click "submit". This view shows all possibler estriction sites but remember you will only
have certain restriction enzymes available to you (given above) Paste in the given enzymes shown above
(at lower left of page) for a custom digest and submit again. You should be able to detect the length of
the fragments that are visible after digestion with this combination enzymes.
2. Using the plasmid maps you have made from above and your lab tech's gel results, you will need to identify the plasmid
with each tube correctly citing the evidence generated from your maps when compared to the gels given above. A written
explanation of the evidence is required for identification of the identity of the plasmid in each tube. (5 points each)
Just identifying the plasmid correcly DOES not earn full credit!
Sequence for pAMP
1 GCGCCCAATA CGCAAACCGC CTCTCCCCGC GCGTTGGCCG ATTCATTAAT GCAGCTGGCA
61 CGACAGGTTT CCCGACTGGA AAGCGGGCAG TGAGCGCAAC GCAATTAATG TGAGTTAGCT
121 CACTCATTAG GCACCCCAGG CTTTACACTT TATGCTTCCG GCTCGTATGT TGTGTGGAAT
181 TGTGAGCGGA TAACAATTTC ACACAGGAAA CAGCTATGAC CATGATTACG CCAAGCTTGC
241 AGAAACGACT TTTTTAAAGG ACGGTTATCA CATTCAAACA TTAATTTTTT ATGATAAACA
301 ATTCCATCCA ATTGATTTAA TCAATACAAC ATTTGAAGAT CAAGCAGATA AATATATTTT
361 TTGGCGTTAT GCAGCTGACA GAGCCAAAAT AACAAATGCC TATGGCTTCA TTTGGATATC
421 AGAGCTATGG CTCAGAAAAG CAAGCATCTA CTCCAATAAA CCAATACATA CAATGCCAAT
481 TATAGATGAA AGACTTCAGG TAATTGGAAT TGATTCAAAT AATAATCAAA AATGTATTTC
541 ATGGAAAATA GTTAGAGAAA ACGAAGAAAA AAAACCGACT TTAGAAATAT CAACAGCAGA
601 CTCAAAACAT GACGAAAAAC CATATTTCAT GCGTTCAGTC TTAAAAGCAA TTGGCGGTGA
661 TGTAAACACT ATGAACAATT GAGTCATAGA ACTTCCATTA TTCTCCTGAA GATAATAATC
721 GCCAAATAAA CCAATACTCA GCTTTACAAT ATACTAACTA ACCGCAGAAC GTTATTTCAT
781 ACAACGTTTC TGCGGCATAT CACAAAACGA TTACTCCATA ACAGGGACAG CAGGCCACTC
841 AATATCAGGT GCAGTTGATG TATCAACACG GTTCAGCAAC ACCCGATACT TCTTCCAGGC
901 TTCCAGCAAC GAGGTTTCTT CCTTCGTTGC AATTTCCAGA TCTGCAGCAT CCTGAAGCGG
961 CGCAATATGC TCACTGGCTA CCTGCATCAG GCTTTTTTTT GTTTCTTCCG CCTCCCGGAT
1021 CCGGAACAGT TTTTCTGCTT CCGTATCCTT CACCCAGGCT GTGCCGTTCC ACTTCTGATA
1081 TTCCCCTCCC GGCGATAACC AGGTAAAATT TTCCGGTAAC GGACCGAGTT CAGAAATAAA
1141 TAACGCGTCG CCGGAAGCCA CGTCATAGAC GGTTTTACCC CGATGGTCTT CAACGAGATG
1201 CCACGATGCC TCATCACTGT TGAAAACAGC CACAAAGCCA GCCGGAATAT CTGGCGGTGC
1261 AATATCGGTA CTGTTTGCAG GCAGACCGGT ATGAGGCGGA ATATATGCGT CACCTTCACC
1321 AATAAATTCA TTAGTTCCGG CCAGCAGATT ATAAATTTTT ATGGTCCGTG GTTGTTCACT
1381 CATTCTGAAT GCCATTATGC AAGCCTCACA ATATAGTTAA ATGCAATGTT TTTGACGGTG
1441 TTTTCCGCGT TACCCGCAGC GTTAACGGTG ATGGTGTGTC CGTGTGAACC AATACTGAAA
1501 GAATGGGCAT GAGCACCGAT AACAACCGGA TGCTGGTGCG CACCAATACC AACTGTATGC
1561 GCATGTGCAC CGGCACTCAC GGCTGTACCG GACAATGAGT GACTGTGGCT GCCCTGACTG
1621 TCCGTTTTCG ATAAATAAGC AATACCCTGT GTGCTGGTTC CTTTAACTGT GGATAAACTT
1681 CCTGTAATGG TTGCTGTTCC ATACTGACTC CAGCCAGAAC TGTTCATCCT TAAACCACTT
1741 GTGTGGGCAT GAGCACCCGC GGCCCCTGTT GAACCGCTCA GACTGTGAGC ATGAGCCCCC
1801 GTGTTATTCG TCGATTTGGT GCCGTAATCG AAACTGCCTG TTGTTTTCGT CCCGTAATCA
1861 AACGACGATG TGGTTTTCGT CCCCAAATCC GTACCGGATG CACTGGCACT GTGGGTGTGC
1921 GACTTAATTC CATCCTGTTC CTGAGACAAT ACAGCACGAC CGCTGGCGGG TTTCCCCTTG
1981 ATTGTCCAGC CTCGCATATC AGGAAGCACA CCCGATGGAT ACGCGACAGC AAGTTTTGGG
2041 TAGGCTGATT TGTCAAACGC CTGCCCCTGC ATCAGGACGT AGCCAGACGG AACGATATCT
2101 GATGGCCACG GGATCGGCGC ACCTGCCGGA AAGGCCGAAT TCACTGGCCG TCGTTTTACA
2161 ACGTCGTGAC TGGGAAAACC CTGGCGTTAC CCAACTTAAT CGCCTTGCAG CACATCCCCC
2221 TTTCGCCAGC TGGCGTAATA GCGAAGAGGC CCGCACCGAT CGCCCTTCCC AACAGTTGCG
2281 CAGCCTGAAT GGCGAATGGC GCCTGATGCG GTATTTTCTC CTTACGCATC TGTGCGGTAT
2341 TTCACACCGC ATATGGTGCA CTCTCAGTAC AATCTGCTCT GATGCCGCAT AGTTAAGCCA
2401 GCCCCGACAC CCGCCAACAC CCGCTGACGC GCCCTGACGG GCTTGTCTGC TCCCGGCATC
2461 CGCTTACAGA CAAGCTGTGA CCGTCTCCGG GAGCTGCATG TGTCAGAGGT TTTCACCGTC
2521 ATCACCGAAA CGCGCGAGAC GAAAGGGCCT CGTGATACGC CTATTTTTAT AGGTTAATGT
2581 CATGATAATA ATGGTTTCTT AGACGTCAGG TGGCACTTTT CGGGGAAATG TGCGCGGAAC
2641 CCCTATTTGT TTATTTTTCT AAATACATTC AAATATGTAT CCGCTCATGA GACAATAACC
2701 CTGATAAATG CTTCAATAAT ATTGAAAAAG GAAGAGTATG AGTATTCAAC ATTTCCGTGT
2761 CGCCCTTATT CCCTTTTTTG CGGCATTTTG CCTTCCTGTT TTTGCTCACC CAGAAACGCT
2821 GGTGAAAGTA AAAGATGCTG AAGATCAGTT GGGTGCACGA GTGGGTTACA TCGAACTGGA
2881 TCTCAACAGC GGTAAGATCC TTGAGAGTTT TCGCCCCGAA GAACGTTTTC CAATGATGAG
2941 CACTTTTAAA GTTCTGCTAT GTGGCGCGGT ATTATCCCGT ATTGACGCCG GGCAAGAGCA
3001 ACTCGGTCGC CGCATACACT ATTCTCAGAA TGACTTGGTT GAGTACTCAC CAGTCACAGA
3061 AAAGCATCTT ACGGATGGCA TGACAGTAAG AGAATTATGC AGTGCTGCCA TAACCATGAG
3121 TGATAACACT GCGGCCAACT TACTTCTGAC AACGATCGGA GGACCGAAGG AGCTAACCGC
3181 TTTTTTGCAC AACATGGGGG ATCATGTAAC TCGCCTTGAT CGTTGGGAAC CGGAGCTGAA
3241 TGAAGCCATA CCAAACGACG AGCGTGACAC CACGATGCCT GTAGCAATGG CAACAACGTT
3301 GCGCAAACTA TTAACTGGCG AACTACTTAC TCTAGCTTCC CGGCAACAAT TAATAGACTG
3361 GATGGAGGCG GATAAAGTTG CAGGACCACT TCTGCGCTCG GCCCTTCCGG CTGGCTGGTT
3421 TATTGCTGAT AAATCTGGAG CCGGTGAGCG TGGGTCTCGC GGTATCATTG CAGCACTGGG
3481 GCCAGATGGT AAGCCCTCCC GTATCGTAGT TATCTACACG ACGGGGAGTC AGGCAACTAT
3541 GGATGAACGA AATAGACAGA TCGCTGAGAT AGGTGCCTCA CTGATTAAGC ATTGGTAACT
3601 GTCAGACCAA GTTTACTCAT ATATACTTTA GATTGATTTA AAACTTCATT TTTAATTTAA
3661 AAGGATCTAG GTGAAGATCC TTTTTGATAA TCTCATGACC AAAATCCCTT AACGTGAGTT
3721 TTCGTTCCAC TGAGCGTCAG ACCCCGTAGA AAAGATCAAA GGATCTTCTT GAGATCCTTT
3781 TTTTCTGCGC GTAATCTGCT GCTTGCAAAC AAAAAAACCA CCGCTACCAG CGGTGGTTTG
3841 TTTGCCGGAT CAAGAGCTAC CAACTCTTTT TCCGAAGGTA ACTGGCTTCA GCAGAGCGCA
3901 GATACCAAAT ACTGTCCTTC TAGTGTAGCC GTAGTTAGGC CACCACTTCA AGAACTCTGT
3961 AGCACCGCCT ACATACCTCG CTCTGCTAAT CCTGTTACCA GTGGCTGCTG CCAGTGGCGA
4021 TAAGTCGTGT CTTACCGGGT TGGACTCAAG ACGATAGTTA CCGGATAAGG CGCAGCGGTC
4081 GGGCTGAACG GGGGGTTCGT GCACACAGCC CAGCTTGGAG CGAACGACCT ACACCGAACT
4141 GAGATACCTA CAGCGTGAGC TATGAGAAAG CGCCACGCTT CCCGAAGGGA GAAAGGCGGA
4201 CAGGTATCCG GTAAGCGGCA GGGTCGGAAC AGGAGAGCGC ACGAGGGAGC TTCCAGGGGG
4261 AAACGCCTGG TATCTTTATA GTCCTGTCGG GTTTCGCCAC CTCTGACTTG AGCGTCGATT
4321 TTTGTGATGC TCGTCAGGGG GGCGGAGCCT ATGGAAAAAC GCCAGCAACG CGGCCTTTTT
4381 ACGGTTCCTG GCCTTTTGCT GGCCTTTTGC TCACATGTTC TTTCCTGCGT TATCCCCTGA
4441 TTCTGTGGAT AACCGTATTA CCGCCTTTGA GTGAGCTGAT ACCGCTCGCC GCAGCCGAAC
4501 GACCGAGCGC AGCGAGTCAG TGAGCGAGGA AGCGGAAGA
pKan DNA sequence:
1 AGCGCCCAAT ACGCAAACCG CCTCTCCCCG CGCGTTGGCC GATTCATTAA TGCAGCTGGC
61 ACGACAGGTT TCCCGACTGG AAAGCGGGCA GTGAGCGCAA CGCAATTAAT GTGAGTTAGC
121 TCACTCATTA GGCACCCCAG GCTTTACACT TTATGCTTCC GGCTCGTATG TTGTGTGGAA
181 TTGTGAGCGG ATAACAATTT CACACAGGAA ACAGCTATGA CCATGATTAC GCCAAGCTTC
241 ACGCTGCCGC AAGCACTCAG GGCGCAAGGG CTGCTAAAGG AAGCGGAACA CGTAGAAAGC
301 CAGTCCGCAG AAACGGTGCT GACCCCGGAT GAATGTCAGC TACTGGGCTA TCTGGACAAG
361 GGAAAACGCA AGCGCAAAGA GAAAGCAGGT AGCTTGCAGT GGGCTTACAT GGCGATAGCT
421 AGACTGGGCG GTTTTATGGA CAGCAAGCGA ACCGGAATTG CCAGCTGGGG CGCCCTCTGG
481 TAAGGTTGGG AAGCCCTGCA AAGTAAACTG GATGGCTTTC TTGCCGCCAA GGATCTGATG
541 GCGCAGGGGA TCAAGATCTG ATCAAGAGAC AGGATGAGGA TCGTTTCGCA TGATTGAACA
601 AGATGGATTG CACGCAGGTT CTCCGGCCGC TTGGGTGGAG AGGCTATTCG GCTATGACTG
661 GGCACAACAG ACAATCGGCT GCTCTGATGC CGCCGTGTTC CGGCTGTCAG CGCAGGGGCG
721 CCCGGTTCTT TTTGTCAAGA CCGACCTGTC CGGTGCCCTG AATGAACTGC AGGACGAGGC
781 AGCGCGGCTA TCGTGGCTGG CCACGACGGG CGTTCCTTGC GCAGCTGTGC TCGACGTTGT
841 CACTGAAGCG GGAAGGGACT GGCTGCTATT GGGCGAAGTG CCGGGGCAGG ATCTCCTGTC
901 ATCTCACCTT GCTCCTGCCG AGAAAGTATC CATCATGGCT GATGCAATGC GGCGGCTGCA
961 TACGCTTGAT CCGGCTACCT GCCCATTCGA CCACCAAGCG AAACATCGCA TCGAGCGAGC
1021 ACGTACTCGG ATGGAAGCCG GTCTTGTCGA TCAGGATGAT CTGGACGAAG AGCATCAGGG
1081 GCTCGCGCCA GCCGAACTGT TCGCCAGGCT CAAGGCGCGC ATGCCCGACG GCGAGGATCT
1141 CGTCGTGACC CATGGCGATG CCTGCTTGCC GAATATCATG GTGGAAAATG GCCGCTTTTC
1201 TGGATTCATC GACTGTGGCC GGCTGGGTGT GGCGGACCGC TATCAGGACA TAGCGTTGGC
1261 TACCCGTGAT ATTGCTGAAG AGCTTGGCGG CGAATGGGCT GACCGCTTCC TCGTGCTTTA
1321 CGGTATCGCC GCTCCCGATT CGCAGCGCAT CGCCTTCTAT CGCCTTCTTG ACGAGTTCTT
1381 CTGAGCGGGA CTCTGGGGTT CGAAATGACC GACCAAGCGA CGCCCAACCT GCCATCACGA
1441 GATTTCGATT CCACCGCCGC CTTCTATGAA AGGTTGGGCT TCGGAATCGT TTTCCGGGAC
1501 GCCGGCTGGA TGATCCTCCA GCGCGGGGAT CTCATGCTGG AGTTCTTCGC CCACCCCGGG
1561 CTCGATCCCC TCGCGAGTTG GTTCAGCTGC TGCCTGAGGC TGGACGACCT CGCGGAGTTC
1621 TACCGGCAGT GCAAATCCGT CGGCATCCAG GAAACCAGCA GCGGCTATCC GCGCATCCAT
1681 GCCCCCGAAC TGCAGGAGTG GGGAGGCACG ATGGCCGCTT TGGTCGACCC GGACGGGACG
1741 CTCCTGCGCC TGATACAGAA CGAATTGCTT GCAGGCATCT CATGAGTGTG TCTTCCCGTT
1801 TTCCGCCTGA GGTCACTGCG TGGATGGAGC GCTGGCGCCT GCTGCGCGAC GGCGAGCTGC
1861 TCACCACCCA CTCGAGCTGG ATACTTCCCG TCCGCCAGGG GGACATGCCG GCGATGCTGA
1921 AGGTCGCGCG CATTCCCGAT GAAGAGGCCG GTTACCGCCT GTTGACCTGG TGGGACGGGC
1981 AGGGCGCCGC CCGAGTCTTC GCCTCGGCGG CGGGCGCTCT GCTCATGGAG CGCGCGTCCG
2041 GGGCCGGGGA CCTTGCACAG ATAGCGTGGT CCGGCCAGGA CGACGAGGCT TGCAGGATCC
2101 CCGGGTACCG AGCTCGAATT CACTGGCCGT CGTTTTACAA CGTCGTGACT GGGAAAACCC
2161 TGGCGTTACC CAACTTAATC GCCTTGCAGC ACATCCCCCT TTCGCCAGCT GGCGTAATAG
2221 CGAAGAGGCC CGCACCGATC GCCCTTCCCA ACAGTTGCGC AGCCTGAATG GCGAATGGCG
2281 CCTGATGCGG TATTTTCTCC TTACGCATCT GTGCGGTATT TCACACCGCA TATGGTGCAC
2341 TCTCAGTACA ATCTGCTCTG ATGCCGCATA GTTAAGCCAG CCCCGACACC CGCCAACACC
2401 CGCTGACGCG CCCTGACGGG CTTGTCTGCT CCCGGCATCC GCTTACAGAC AAGCTGTGAC
2461 CGTCTCCGGG AGCTGCATGT GTCAGAGGTT TTCACCGTCA TCACCGAAAC GCGCGAGACG
2521 AAAGGGCCTC GTGATACGCC TATTTTTATA GGTTAATGTC ATGATAATAA TGGTTTCTTA
2581 GACGTCAGGT GGCACTTTTC GGGGAAATGT GCGCGGAACC CCTATTTGTT TATTTTTCTA
2641 AATACATTCA AATATGTATC CGCTCATGAG ACAATAACCC TGATAAATGC TTCAATAATA
2701 CTCACCAGTC ACAGAAAAGC ATCTTACGGA TGGCATGACA GTAAGAGAAT TATGCAGTGC
2761 TGCCATAACC ATGAGTGATA ACACTGCGGC CAACTTACTT CTGACAACGA TCGGAGGACC
2821 GAAGGAGCTA ACCGCTTTTT TGCACAACAT GGGGGATCAT GTAACTCGCC TTGATCGTTG
2881 GGAACCGGAG CTGAATGAAG CCATACCAAA CGACGAGCGT GACACCACGA TGCCTGTAGC
2941 AATGGCAACA ACGTTGCGCA AACTATTAAC TGGCGAACTA CTTACTCTAG CTTCCCGGCA
3001 ACAATTAATA GACTGGATGG AGGCGGATAA AGTTGCAGGA CCACTTCTGC GCTCGGCCCT
3061 TCCGGCTGGC TGGTTTATTG CTGATAAATC TGGAGCCGGT GAGCGTGGGT CTCGCGGTAT
3121 CATTGCAGCA CTGGGGCCAG ATGGTAAGCC CTCCCGTATC GTAGTTATCT ACACGACGGG
3181 GAGTCAGGCA ACTATGGATG AACGAAATAG ACAGATCGCT GAGATAGGTG CCTCACTGAT
3241 TAAGCATTGG TAACTGTCAG ACCAAGTTTA CTCATATATA CTTTAGATTG ATTTAAAACT
3301 TCATTTTTAA TTTAAAAGGA TCTAGGTGAA GATCCTTTTT GATAATCTCA TGACCAAAAT
3361 CCCTTAACGT GAGTTTTCGT TCCACTGAGC GTCAGACCCC GTAGAAAAGA TCAAAGGATC
3421 TTCTTGAGAT CCTTTTTTTC TGCGCGTAAT CTGCTGCTTG CAAACAAAAA AACCACCGCT
3481 ACCAGCGGTG GTTTGTTTGC CGGATCAAGA GCTACCAACT CTTTTTCCGA AGGTAACTGG
3541 CTTCAGCAGA GCGCAGATAC CAAATACTGT CCTTCTAGTG TAGCCGTAGT TAGGCCACCA
3601 CTTCAAGAAC TCTGTAGCAC CGCCTACATA CCTCGCTCTG CTAATCCTGT TACCAGTGGC
3661 TGCTGCCAGT GGCGATAAGT CGTGTCTTAC CGGGTTGGAC TCAAGACGAT AGTTACCGGA
3721 TAAGGCGCAG CGGTCGGGCT GAACGGGGGG TTCGTGCACA CAGCCCAGCT TGGAGCGAAC
3781 GACCTACACC GAACTGAGAT ACCTACAGCG TGAGCTATGA GAAAGCGCCA CGCTTCCCGA
3841 AGGGAGAAAG GCGGACAGGT ATCCGGTAAG CGGCAGGGTC GGAACAGGAG AGCGCACGAG
3901 GGAGCTTCCA GGGGGAAACG CCTGGTATCT TTATAGTCCT GTCGGGTTTC GCCACCTCTG
3961 ACTTGAGCGT CGATTTTTGT GATGCTCGTC AGGGGGGCGG AGCCTATGGA AAAACGCCAG
4021 CAACGCGGCC TTTTTACGGT TCCTGGCCTT TTGCTGGCCT TTTGCTCACA TGTTCTTTCC
4081 TGCGTTATCC CCTGATTCTG TGGATAACCG TATTACCGCC TTTGAGTGAG CTGATACCGC
4141 TCGCCGCAGC CGAACGACCG AGCGCAGCGA GTCAGTGAGC GAGGAAGCGG AAGA
pGlo DNA Sequence
1 atcgatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac
61 tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca
121 ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta
181 aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata
241 ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag
301 cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag
361 caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg
421 tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct
481 tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc
541 ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc
601 gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca
661 tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga
721 tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa
781 acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata
841 taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc
901 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt
961 tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat
1021 tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta
1081 accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt
1141 aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca
1201 ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta
1261 tcgcaactct ctactgtttc tccatacccg tttttttggg ctagaaataa ttttgtttaa
1321 ctttaagaag gagatataca tatggctagc aaaggagaag aacttttcac tggagttgtc
1381 ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt cagtggagag
1441 ggtgaaggtg atgctacata cggaaagctt acccttaaat ttatttgcac tactggaaaa
1501 ctacctgttc catggccaac acttgtcact actttctctt atggtgttca atgcttttcc
1561 cgttatccgg atcatatgaa acggcatgac tttttcaaga gtgccatgcc cgaaggttat
1621 gtacaggaac gcactatatc tttcaaagat gacgggaact acaagacgcg tgctgaagtc
1681 aagtttgaag gtgataccct tgttaatcgt atcgagttaa aaggtattga ttttaaagaa
1741 gatggaaaca ttctcggaca caaactcgag tacaactata actcacacaa tgtatacatc
1801 acggcagaca aacaaaagaa tggaatcaaa gctaacttca aaattcgcca caacattgaa
1861 gatggatccg ttcaactagc agaccattat caacaaaata ctccaattgg cgatggccct
1921 gtccttttac cagacaacca ttacctgtcg acacaatctg ccctttcgaa agatcccaac
1981 gaaaagcgtg accacatggt ccttcttgag tttgtaactg ctgctgggat tacacatggc
2041 atggatgagc tctacaaata atgaattcga gctcggtacc cggggatcct ctagagtcga
2101 cctgcaggca tgcaagcttg gctgttttgg cggatgagag aagattttca gcctgataca
2161 gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc
2221 ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag
2281 tgtggggtcc cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca
2341 gtgcaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag
2401 gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc
2461 aggacgcccg ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg
2521 cctttttgcg tttctacaaa ctctttgttt atttttctaa atacattcaa atatgtatcc
2581 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
2641 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
2701 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
2761 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
2821 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
2881 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
2941 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
3001 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
3061 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
3121 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
3181 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
3241 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
3301 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
3361 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
3421 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
3481 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttacg
3541 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
3601 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
3661 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
3721 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
3781 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
3841 tcttgttcca aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag
3901 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
3961 cgaattttaa caaaatatta acgtttacaa tttaaaagga tctaggtgaa gatccttttt
4021 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
4081 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
4141 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
4201 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
4261 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
4321 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
4381 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
4441 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagcattga
4501 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
4561 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
4621 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
4681 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
4741 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
4801 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
4861 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
4921 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
4981 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
5041 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
5101 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
5161 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
5221 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
5281 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
5341 aggatctaat tctcatgttt gacagcttat c