LOCUS       sequence-313905-        9331 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Allows for constituitive mammalian expression of the HCoV-NL63 3CL 
            protease with C144A mutation via transfection or lentivirus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9331)
  AUTHORS   Resnick SJ, Iketani S, Hong SJ, Zask A, Liu H, Kim S, Melore S, Lin 
            FY, Nair MS, Huang Y, Lee S, Tay NES, Rovis T, Yang HW, Xing L, 
            Stockwell BR, Ho DD, Chavez A
  TITLE     Inhibitors of Coronavirus 3CL Proteases Protect Cells from 
            Protease-Mediated Cytotoxicity.
  JOURNAL   J Virol. 2021 Jun 24;95(14):e0237420. doi: 10.1128/JVI.02374-20. 
            Epub 2021 Jun 24.
  PUBMED    33910954
REFERENCE   2  (bases 1 to 9331)
  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: "J Virol."; 
            date: "2021-06-24"; volume: "95(14):e0237420. doi"; pages: " 
            10.1128/JVI.02374-20. Epub 2021 Jun 24"
COMMENT     Sequence Label: pLEX307-HCoV-NL63-3CL-C144A
FEATURES             Location/Qualifiers
     source          1..9331
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        96..1274
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          327..1265
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1222..1242
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1294..1318
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2240..2264)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2272..2313
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    2364..2952
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2417..2437)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2835..2846)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3024..3257
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3335..3456
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3372..3391)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3426..3445
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3496..3631
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3651..3670)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3652..3670)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3680..3697)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3680..3696)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3689..3711)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3838..4293
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3925..3944)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4135..4156
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4319..4423
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4424..4492,4493..5284)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4642..4661)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5455..6043
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5944..5963
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6197..6214
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6331..6352
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6367..6372,6373..6390,6391..6397)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6405..6421
                     /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     6410..6432
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6429..6445
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6429..6445
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6464..6484
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6466..6484
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6512..6738
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6739..6919
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6966..7091
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7584..7817
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8002..8046
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        8277..8487
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        8299..8495
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8346..8481
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             8527..9129
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGAS"
     primer_bind     complement(8527..8546)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9023..9043
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9172..9289
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ctcgaggcct gcaggtgcaa agatggataa agttttaaac agagaggaat ctttgcagct
       61 aatggacctt ctaggtcttg aaaggagtgg gaattggctc cggtgcccgt cagtgggcag
      121 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg
      181 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
      241 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
      301 gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc
      361 tctttacggg ttatggccct tgcgtgcctt gaattacttc cacctggctg cagtacgtga
      421 ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag
      481 gagccccttc gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc
      541 gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa
      601 atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc
      661 aagatctgca cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt
      721 cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg
      781 ggtagtctca agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc
      841 cgccctgggc ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc
      901 ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg
      961 gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact
     1021 ccacggagta ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt tggagtacgt
     1081 cgtctttagg ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg
     1141 agactgaagt taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg
     1201 agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat
     1261 ttcaggtgtc gtgaggctag catcgattga tcaacaagtt tgtacaaaaa agcaggctac
     1321 aaaatgtctg gtcttaagaa gatggcacaa ccatctggtt gtgttgagag atgtgtggtt
     1381 cgcgtctgtt atggtagtac tgtgcttaat ggagtttggt taggtgacac tgttacttgt
     1441 cctagacatg tcatagcacc atcaaccact gttcttattg attatgatca tgcatatagt
     1501 actatgcgtt tgcataattt ttcagtgtct tataatggtg tcttcttggg agttgttggt
     1561 gttacaatgc atggttctgt gttgcgtatt aaggtttcac aatctaatgt acatacacct
     1621 aaacatgttt ttaaaacgtt gaaacctggt gattctttta atattttagc atgttatgaa
     1681 ggtattgcat ctggtgtttt tggtgttaat ttacgtacaa actttactat taaaggttct
     1741 tttataaatg gagctgccgg ttctcctggt tataatgtta gaaatgatgg tactgttgag
     1801 ttttgttatt tacaccaaat tgagttaggt agtggtgctc atgttggttc tgattttact
     1861 ggtagtgttt atggtaattt tgatgaccaa cctagtttgc aagttgagag tgccaacctt
     1921 atgctatcag ataatgttgt tgcctttttg tatgctgctt tgttgaatgg ttgtaggtgg
     1981 tggttgcgtt caactagagt taatgttgat ggttttaatg aatgggctat ggctaatggt
     2041 tatacaagtg tttctagtgt tgagtgctat tctattttgg cagcaaaaac tggtgttagt
     2101 gttgaacaat tgttagcttc cattcaacat cttcatgaag gttttggtgg taaaaacata
     2161 cttggttatt ctagtttatg tgatgagttc acactagctg aagttgtgaa gcagatgtat
     2221 ggtgttaact tgcaataaaa cccagctttc ttgtacaaag tggttgatat cggtaagcct
     2281 atccctaacc ctctcctcgg tctcgattct acgtagtaat gaactagtac cggttaagtc
     2341 gacaatcaac gcgttaagtc gacaatcaac ctctggatta caaaatttgt gaaagattga
     2401 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     2461 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     2521 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     2581 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     2641 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     2701 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     2761 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     2821 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     2881 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     2941 ccgcctcccc gcgtcgactt taagaccaat gacttacaag gcagctgtag atcttagcca
     3001 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct
     3061 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     3121 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     3181 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     3241 gtggaaaatc tctagcagta cgtatagtag ttcatgtcat cttattattc agtatttata
     3301 acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg
     3361 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3421 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg
     3481 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3541 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3601 cagaagtagt gaggaggctt ttttggaggc ctagggacgt acccaattcg ccctatagtg
     3661 agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     3721 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     3781 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg
     3841 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3901 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3961 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4021 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     4081 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4141 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     4201 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4261 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     4321 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4381 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4441 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4501 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4561 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4621 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4681 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4741 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4801 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4861 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4921 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4981 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5041 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5101 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5161 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5221 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5281 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5341 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5401 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5461 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5521 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5581 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5641 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5701 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5761 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5821 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5881 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5941 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6001 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6061 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6121 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     6181 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     6241 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     6301 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     6361 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     6421 atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta
     6481 aagggaacaa aagctggagc tgcaagctta atgtagtctt atgcaatact cttgtagtct
     6541 tgcaacatgg taacgatgag ttagcaacat gccttacaag gagagaaaaa gcaccgtgca
     6601 tgccgattgg tggaagtaag gtggtacgat cgtgccttat taggaaggca acagacgggt
     6661 ctgacatgga ttggacgaac cactgaattg ccgcattgca gagatattgt atttaagtgc
     6721 ctagctcgat acataaacgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     6781 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     6841 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     6901 gtgtggaaaa tctctagcag tggcgcccga acagggactt gaaagcgaaa gggaaaccag
     6961 aggagctctc tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc
     7021 ggcgactggt gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt
     7081 gcgagagcgt cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa
     7141 ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag
     7201 aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg
     7261 gacagctaca accatccctt cagacaggat cagaagaact tagatcatta tataatacag
     7321 tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag
     7381 acaagataga ggaagagcaa aacaaaagta agaccaccgc acagcaagcg gccgctgatc
     7441 ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa
     7501 gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag
     7561 agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga
     7621 agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt
     7681 atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa
     7741 ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta
     7801 aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct
     7861 gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc
     7921 tggatggagt gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa
     7981 gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca
     8041 agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg
     8101 atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga
     8161 gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc
     8221 atgcattgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     8281 gcagaagtat gcaaagcatg cgtctcaatt agtcagcaac catagtcccg cccctaactc
     8341 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     8401 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     8461 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct ttctagaggt accaccggag
     8521 cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
     8581 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
     8641 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     8701 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg ccgtggcggt ctggaccacg
     8761 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     8821 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
     8881 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     8941 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
     9001 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     9061 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     9121 gccagttaac gaattctaga tcttgagaca aatggcagta ttcatccaca attttaaaag
     9181 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
     9241 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta
     9301 cagggacagc agagatccac tttggcgccg g
//
