LOCUS       sequence-428833-        9765 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a single point mutant of melanoma 
            differentiation-associated protein 5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9765)
  TITLE     MDA5 plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9765)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pET50_G495R MDA5
FEATURES             Location/Qualifiers
     source          1..9765
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1..48)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     44..62
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(143..187)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(188..205)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(3320..3343)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(3353..3370)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(3386..4870)
                     /codon_start=1
                     /gene="nusA"
                     /product="transcription elongation protein (N utilization 
                     substance protein A)"
                     /label=NusA
                     /note="highly soluble in E. coli"
                     /translation="MNKEILAVVEAVSNEKALPREKIFEALESALATATKKKYEQEIDV
                     RVQIDRKSGDFDTFRRWLVVDEVTQPTKEITLEAARYEDESLNLGDYVEDQIESVTFDR
                     ITTQTAKQVIVQKVREAERAMVVDQFREHEGEIITGVVKKVNRDNISLDLGNNAEAVIL
                     REDMLPRENFRPGDRVRGVLYSVRPEARGAQLFVTRSKPEMLIELFRIEVPEIGEEVIE
                     IKAAARDPGSRAKIAVKTNDKRIDPVGACVGMRGARVQAVSTELGGERIDIVLWDDNPA
                     QFVINAMAPADVASIVVDEDKHTMDIAVEAGNLAQAIGRNGQNVRLASQLSGWELNVMT
                     VDDLQAKHQAEAHAAIDTFTKYLDIDEDFATVLVEEGFSTLEELAYVPMKELLEIEGLD
                     EPTVEALRERAKNALATIAQAQEESLGDNKPADDLLNLEGVDRDLAFKLAARGVCTLED
                     LAEQGIDDLADIEGLTDEKAGALIMAARNICWFGDEA"
     CDS             complement(4877..4894)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             4914..4936
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4951..4975
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     complement(4975..4994)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4976..4994)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5082..5101)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5307..5384
                     /gene="lacI"
                     /label=lacI promoter
     CDS             5385..6467
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(5404..5423)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             7039..7230
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(7224..7246)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    7332..7471
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     7387..7406
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7486..7503)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7657..8245)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7737..7756)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(8356..8375)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             8367..9182
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             8367..9182
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(8440..8459)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(9274..9729)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9411..9432)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9623..9642
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctagtt attgctcagc
       61 ggtggcagca gccaactcag cttcctttcg ggctttgttt agcagcctag gttaattaag
      121 cctcgagagc agcagaagta gagctgtcca tgtgctggcg ttcgaattta gcagcagcgg
      181 tttctttact accgcgtggc accagagcga gctctgcggc cgcaagcttg tcgacctaat
      241 cctcatcact aaataaacag cattctgaat agtcaagatt gggaaatgtg ataggtaatt
      301 ctacccactt tttgtattgt ttctttgttg aattattttt gaaaaccact acaaaattcc
      361 ttattttgag acaaggcaaa tctaagcctt tgtgcaccat cattgttccc caagcctggc
      421 cacatttgca gatgatttca ccatttattt gatagtcggc acacttcttt tgcagtgttt
      481 tgttttctct tacaatgtaa agttccttga attctggggt catattgacg tgatgcattt
      541 tctcaattac atggatatct tccccagaac aggctagcac actgcagttt ttgcaaagga
      601 aagttattag tgatgggtta ttcttgtaat gcttggcaat atttctcttg gttttcattt
      661 tcttttccat tatactttgc atctgtaatt ccaaaatctt atgagcatac tcctctggtt
      721 tcatattttg aacacaatgt atagctttat acatcatctt ctctcggaaa tcattaactg
      781 tctcacgttc gataactcct gaaccactgt gagcaaccag gacgtaggtg ctctcatcag
      841 ctctggctcg accacgggcc tggaccatgg ctatttcatt ggtgacgaga ccataacgga
      901 taacaatgtt acattcttta atatccagac cttcttctgc cactgtggta gcgataagca
      961 gatttatttt tccagtgcga aatttactaa tgacttcttt ttgttcattc tgtgtcatgg
     1021 gtttgaactc actgctgtgt ccagctccaa tcagatggtg ggctttgact cctacttcag
     1081 caaatttttc attttcagta atccactggg aaagcgcata tgcactctgt cgtgtttttg
     1141 taaagattat tcctcgtgct gattcctcag tcctagtata ttgctccatt atggtatttc
     1201 ttaatttggt cagcttttca ttttcatatt ctgggttttc agccagcctt ttcaacattt
     1261 tattgttttc aaaaaataaa gtcatgagaa atctatctgt ttcatccagt ttcaaaggtt
     1321 tctttaaatc atcctcatct tcatcaccat cacaatactc atcatcacca ccctcatcac
     1381 tatcatcttc tatgactgca aacttcttat ctttctcttc attatagaaa gtttcaagat
     1441 gagtatacgc atctatcatt cgaattgtgt cattaatttg tagggcctca ttgtacttcc
     1501 tcaaatgttc tgcacaaaca cgttctttgc gatttccttc ttttgcagct tttttttcca
     1561 tttgaatggc ccattgttca tagggttgag ttccaaaatc tgacattgga ctcatttgac
     1621 aataagtttg aatccttgtc attatttcta gaagtttctc tttaaatgga tcttctctgg
     1681 ttgcatctgc aatggcaaac ttcttgcatg gctcctgtat ttggtttttc agttgatcaa
     1741 ggttttcttt aacagtttta atagtaaatg catcaagatt ggcacatagt tttaaaatgt
     1801 gttcttcagc tttggcttgc ttcgtggccc ttccaacacc aggtgaagct gttagtccca
     1861 gtatctgagg aaggggaatc actggtttgt tttctttctt gagtctattg tttttcaact
     1921 tctgcatcaa ataatgcctc atgatgttat tatacactgc ttctttgttg gtgtgatgac
     1981 attcatcaat gataatgagg gaaaagtctg acaattgaac accagcatct tctccatttt
     2041 ccaagtttaa gagggagttt tcaaggattt gagctgtact gataataata tcacaggact
     2101 tgacaacttc tggaaatgat attttcagtt gggtatcacc acttaatcca ataacacgat
     2161 accatttctt caaaaatggt tggaactcct tgcggaagag ctgttcaact agcagtacct
     2221 tattgacaag aactataact tttccaggct cagatgcttt tttcttcttg tctaagtgat
     2281 ccttggcaat gtaaacagcc actctggttt ttccactccc tgtagggagg cagatgatga
     2341 tattcttccc ttccaaggct ggctgggcaa cttccatttg gtaaggcctg agctggagtt
     2401 ctggctccgg ggatgctctt gctgccacat tctcttcatc tgaatcactt cccatggtgc
     2461 ctgaatcact gcccatgttg ctgttatgtc caagactttc atctaagcag ctgacacttc
     2521 cttctgccaa acttgtgtct gattctgaaa ctacagaaga atctgcaaaa gatgattctg
     2581 atgagttatt ctccatgccc cagacctcct tctccagatt tggctgaact gtggttgaaa
     2641 gaagttgctc ttccacttga ggaccatcaa cttgtgataa attctcaatc tctgcattgc
     2701 tttctgagca atcagagcct gttaactctt ggacaagttc attgtttcct gtttgacgaa
     2761 gaacattcag aaatgcagag aaccagtttt ctttctgcac aatccttttt agtagctctc
     2821 ttacacctga ttcatttcca ttgttttctg cagcagcaat ccggtttctg tcttcaattg
     2881 tcaacagttc ctcctccatg cacttatcca agacgtctct aactagaagc ttgtccacca
     2941 gagtgggctg aaggaggttc agcagttgga gatattcatc atgagcgttc tcaaacgatg
     3001 gagagggcaa gtccgtgagc tcagggttca tgtagcgggc ggccagaggg ctgccggttc
     3061 tccggagggc ctccacgaat tcccgagtcc aaccaaggtg ccagactccc ttctccaagg
     3121 tgctcagcag cagttcaact gcctgcatgt tcccggaggt ggcgactgtc ctctgaatct
     3181 gctccttcac ctctgcaggc agaaaggtca ggtagtccag cacaggctcc acctggatgt
     3241 acattttcac cctggccctg aagcacgaga tgagatagcg gaaattctcg tctgtggaat
     3301 acccattcga catagccccg ggtccctgaa agaggacttc aagagccgcg gagtgatggt
     3361 gatggtgatg accagaacca ctagtcgctt cgtcaccgaa ccagcaaata ttacgggcag
     3421 ccataatcag tgctccggct ttttcgtcgg tcaacccttc gatatcagcc agatcatcaa
     3481 tgccctgttc ggcgagatct tccagcgtac aaacgccacg ggcggccagt ttgaatgcca
     3541 aatcacgatc taccccttca aggttcagca gatcgtcagc cggtttgtta tcaccgaggc
     3601 tttcttcctg ggcctgtgca atggtggcca gtgcattttt agcacgctcg cgcagtgctt
     3661 caacggtcgg ctcatcaagg ccttcgattt ccaacagctc tttcatcggc acataggcca
     3721 attcttccag cgtcgagaag ccttcttcta ccagaacagt cgcgaagtct tcgtcgatgt
     3781 cgagatattt ggtgaaggtg tcgatcgctg cgtgcgcttc cgcctgatgc ttagcttgca
     3841 ggtcgtcaac ggtcatcacg ttgagttccc aaccgctcag ttgcgaagcc agacgcacgt
     3901 tctgaccgtt acggccaatc gcctgcgcca gattaccggc ttcaacggcg atgtccatgg
     3961 tgtgtttatc ttcatccacc acgatagaag caacgtctgc cggtgccatt gcgttaatca
     4021 cgaactgcgc cgggttatca tcccacagga cgatatcgat acgctcgcca cccagttcag
     4081 tagacaccgc ctgaacacgc gcgccacgca tacctacgca agcacctacc ggatcgatac
     4141 gtttatcgtt ggttttcacc gcgattttcg cacgagaacc cggatcgcga gccgctgctt
     4201 taatttcaat cacttcttcg ccgatttctg gcacttcaat acggaacagt tcgatcagca
     4261 tttccggctt ggaacgagtg acgaacagtt gcgcgccacg cgcttccggg cgaacggaat
     4321 agagcacgcc acgaacgcgg tcgccagggc ggaagttttc acgcggcagc atatcttcgc
     4381 gcaggatcac ggcttcagcg ttgttgccca gatccagaga gatgttgtcg cggtttactt
     4441 ttttcaccac gccggtgatg atttcacctt cgtgttcacg gaactgatca accaccatcg
     4501 cacgttcggc ttcacgcact ttctgcacga taacctgttt tgccgtctgg gtagtgatac
     4561 ggtcaaaggt aacagactca atctgatctt caacgtaatc gcccaggttc aggctttcat
     4621 cttcataacg tgcggcttca agggtgattt ccttggtcgg ctgggtgact tcatcaacaa
     4681 ctaaccagcg acggaaagtg tcaaaatcac cgcttttgcg atcgatctgt acgcggacgt
     4741 cgatctcttg ttcatatttt ttctttgttg ctgtcgccag cgcgctttcc aatgcttcga
     4801 aaatcttctc gcgaggtagc gccttttcat tggatacggc ttcaactaca gccaaaattt
     4861 ctttgttcat actagaatgg tgatggtggt gatggctgct gcccatatgt atatctcctt
     4921 cttaaagtta aacaaaatta tttctagagg ggaattgtta tccgctcaca attcccctat
     4981 agtgagtcgt attaatttcg cgggatcgag atcgatctcg atcctctacg ccggacgcat
     5041 cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac
     5101 cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat
     5161 ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg caccattcct
     5221 tgcggcggcg gtgctcaacg gcctcaacct actactgggc tgcttcctaa tgcaggagtc
     5281 gcataaggga gagcgtcgag atcccggaca ccatcgaatg gcgcaaaacc tttcgcggta
     5341 tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt
     5401 tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc
     5461 aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga
     5521 attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg
     5581 ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc
     5641 gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag
     5701 cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact
     5761 atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt
     5821 tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg
     5881 gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag
     5941 cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca
     6001 ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt
     6061 ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca
     6121 acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg
     6181 cggacatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt
     6241 taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc
     6301 aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa
     6361 gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     6421 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     6481 aatgtaagtt agctcactca ttaggcaccg ggatctcgac cgatgccctt gagagccttc
     6541 aacccagtca gctccttccg gtgggcgcgg ggcatgacta gcatgatcgt gctcctgtcg
     6601 ttgaggaccc ggctaggctg gcggggttgc cttactggtt agcagaatga atcaccgata
     6661 cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc aacaacatga
     6721 atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc gccctgcacc
     6781 attatgttcc ggatctgcat cgcaggatgc tgctggctac cctgtggaac acctacatct
     6841 gtattaacga agcgctggca ttgaccctga gtgatttttc tctggtcccg ccgcatccat
     6901 accgccagtt gtttaccctc acaacgttcc agtaaccggg catgttcatc atcagtaacc
     6961 cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta cccccatgaa cagaaatccc
     7021 ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt
     7081 atcagaagcc agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag
     7141 gcagacatct gtgaatcgct tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg
     7201 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     7261 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     7321 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta
     7381 actatgcggc atcagagcag attgtactga gagtgcacca ttgcggtgtg aaataccgca
     7441 cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc
     7501 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     7561 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     7621 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     7681 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     7741 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7801 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7861 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7921 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7981 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     8041 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     8101 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     8161 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     8221 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     8281 tcagtggaac gaaaactcac gttaagggat tttggtcatg aacaataaaa ctgtctgctt
     8341 acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg tcttgctcta
     8401 ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata
     8461 atgtcgggca atcaggtgcg acaatctatc gattgtatgg gaagcccgat gcgccagagt
     8521 tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac
     8581 taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg
     8641 atgatgcatg gttactcacc actgcgatcc ccggcaaaac agcattccag gtattagaag
     8701 aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc
     8761 attcgattcc tgtttgtaat tgtcctttta acagtgatcg cgtatttcgt ctcgctcagg
     8821 cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg
     8881 gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt
     8941 cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa
     9001 taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc
     9061 tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg
     9121 gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct
     9181 aagaattaat tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     9241 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa
     9301 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     9361 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     9421 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     9481 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     9541 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     9601 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     9661 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     9721 agggcgcgtc ccattcgcca atccggatat agttcctcct ttcag
//
