LOCUS       sequence-479628-       11477 bp    DNA     circular SYN 24-JUL-2025
DEFINITION  mammalian expression of MERS-CoV soluble spike trimer protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11477)
  AUTHORS   Esposito D, Mehalko J, Drew M, Snead K, Wall V, Taylor T, Frank P, 
            Denson JP, Hong M, Gulten G, Sadtler K, Messing S, Gillette W
  TITLE     Optimizing high-yield production of SARS-CoV-2 soluble spike trimers
            for serology assays.
  JOURNAL   Protein Expr Purif. 2020 Jun 4;174:105686. doi: 
            10.1016/j.pep.2020.105686.
  PUBMED    32504802
REFERENCE   2  (bases 1 to 11477)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 24, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Protein 
            Expr Purif."; date: "2020-06-4"; volume: "174:105686. doi"; pages: "
            10.1016/j.pep.2020.105686"
COMMENT     Sequence Label: R619-M88-303: CMV51p> MERS-CoV
            S-2P-T4f-3C-His...
FEATURES             Location/Qualifiers
     source          1..11477
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     505..529
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    574..598
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1248..1259
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             4570..4593
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4597..4620
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             4627..4650
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4690..4713
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    complement(4715..4739)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(4801..4819)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4802..4819)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4839..4856)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4845..5069
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5115..5543
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5202..5221)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5412..5433
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5552..5572)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5557..5886
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5737..5872
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5799..5818
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5953..6747
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6007..6026)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6617..6636
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6921..7042
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6958..6977)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7012..7031
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7091..7107)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7091..7107)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7104..7126)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7115..7131
                     /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)."
     promoter        complement(join(7139..7145,7146..7163,7164..7169))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7184..7205
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7322..7339)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7493..8081)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7573..7592)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8252..9043,9044..9112))
                     /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     8875..8894
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9113..9217)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      9452..11241
                     /label=oriP
                     /note="Epstein-Barr virus oriP replication origin (Yates et
                     al., 2000)"
     primer_bind     complement(11278..11297)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    11447..11467
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtccgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta cgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacaccaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      541 ccatccacgc tgttttgacc tccatagaag acaacaactt tgtacaaaaa agttggcacc
      601 atgattcaca gcgtgttcct cctgatgttc cttctgaccc ccaccgagag ctacgtggac
      661 gtcgggccag atagcgtgaa gtcggcgtgt atcgaggtgg acatccagca gaccttcttc
      721 gataagacct ggcctaggcc catcgacgtg tccaaggcgg atggcatcat ctacccgcag
      781 ggtcggactt actccaacat cactattacc taccaaggcc tgttcccata tcaaggagat
      841 cacggcgata tgtacgtcta ctcggctgga catgcgaccg gcacaacccc ccagaaactg
      901 tttgtggcca actactccca ggacgtgaag cagttcgcca atggtttcgt cgtgcggatt
      961 ggagcggccg ccaactcgac cggaaccgtg attattagcc ctagcacctc cgccaccatc
     1021 aggaagatct acccggcctt tatgcttggg tcgagcgtgg gaaacttcag cgacggcaaa
     1081 atgggccgct tcttcaacca taccctcgtg ctgctgcctg acggctgcgg taccctcctc
     1141 cgcgccttct actgtatcct ggagcctaga tccggcaacc attgcccggc tgggaactca
     1201 tacacctcct tcgcaactta ccacactcca gccaccgatt gctcagatgg gaactacaac
     1261 cggaatgcgt cgctgaactc gttcaaggaa tacttcaact tgcgaaactg cactttcatg
     1321 tacacctaca acatcactga ggacgaaatt ctcgagtggt ttggtattac tcagaccgcc
     1381 cagggagtgc atctgttctc ctcgcgctat gtcgacctat acggtggaaa catgttccag
     1441 ttcgccacac ttcctgtgta tgacaccatt aagtactatt caatcattcc tcactcgatt
     1501 agatctatcc agtccgacag aaaggcctgg gccgcattct acgtgtacaa gctgcagcct
     1561 ctgaccttcc tgctggattt tagcgtggac ggctacatca gacgcgccat tgactgcggc
     1621 ttcaatgacc tgagccagct gcactgctcg tacgagtctt tcgatgtgga gtcaggcgta
     1681 tactcagttt cgtctttcga agcgaagccg agcggttccg tcgtcgaaca agccgaaggg
     1741 gtggaatgcg acttcagccc cttgttgagc gggactcccc cacaagtgta caacttcaag
     1801 cgcctggtgt tcactaactg caactataac ctgactaagc tgctttcgct gttctccgtg
     1861 aacgacttta cctgttccca aatctcccct gccgctatcg cctccaactg ttactcaagc
     1921 ctcatcctgg attacttctc ctacccactg tcgatgaaga gtgacctctc ggtgtcctct
     1981 gccggaccga tcagccagtt caattacaag cagagcttta gcaaccccac ttgcctcatt
     2041 ctcgcaaccg tgccccacaa cctgaccacg attactaagc ccctgaagta ctcatatatc
     2101 aacaagtgct cccgcttcct ctccgacgac cgcactgaag tcccgcagct cgtcaacgca
     2161 aaccagtact ccccctgcgt gtccatcgtg ccctccaccg tgtgggaaga tggggattac
     2221 taccggaagc agctgtcacc gctcgaaggc ggaggatggc ttgtggcctc cggaagcacc
     2281 gtggccatga ccgaacagct gcagatgggg ttcggcatca ctgtgcagta cggaacggac
     2341 accaactccg tatgccccaa gctggagttc gccaacgaca cgaagattgc tagccagctc
     2401 ggcaactgcg tggagtactc actgtacggg gtgtcaggac gcggcgtgtt ccagaactgc
     2461 accgccgtgg gagtcagaca gcagcgcttc gtgtacgacg cctaccaaaa cctggtcgga
     2521 tactactctg acgacgggaa ctactactgc ctgcgggctt gcgtgtccgt gccggtgtcc
     2581 gtgatctacg acaaggagac aaagacccac gccaccctct ttggatcagt ggcctgcgag
     2641 cacatctcgt caactatgtc gcaatactcg cggtccacac ggtccatgct gaagcgccgc
     2701 gattcgacct atggaccgct gcagactcct gtgggctgtg tgcttggact cgtgaacagc
     2761 tccctgttcg tggaggattg caagctgccg cttggacaaa gtctgtgcgc gctgccagat
     2821 actccgtcca ctctgacccc tgcttccgtc ggatccgtgc ccggagaaat gcggctggcg
     2881 tccattgcct tcaaccaccc gatccaagtg gaccagctga actcctcata ctttaagttg
     2941 agcatcccta cgaatttctc gttcggcgtg acccaggagt acatccagac tactattcaa
     3001 aaggtcaccg tggactgcaa gcagtatgtc tgcaacggat tccagaagtg cgagcagctg
     3061 ctgagagaat atgggcagtt ttgctccaaa atcaaccagg ccctccatgg agccaacttg
     3121 cggcaggacg attccgtcag gaacctgttc gcgtccgtga agtcctccca gtcgagccca
     3181 atcatcccgg gattcggcgg agatttcaac ctgacgcttc tggaaccggt gtccatttcc
     3241 accgggagca gatcggccag aagcgccatt gaagatctgc tgttcgataa ggtcaccata
     3301 gcagaccctg gctacatgca gggatacgac gactgcatgc agcagggtcc ggctagcgca
     3361 agagatctga tctgcgcgca atacgtggcg ggctacaagg tcctgccgcc tctcatggac
     3421 gtgaatatgg aggcggctta cacctcctct ctgctcgggt ccatcgcggg agtgggatgg
     3481 actgctggcc tttcgtcgtt cgccgccatt ccgttcgccc agtccatttt ctaccggctg
     3541 aacggcgtcg gaatcactca acaagtcctc tccgaaaacc agaagctgat tgccaacaag
     3601 ttcaaccagg cactgggagc catgcagacc gggttcacca ccacgaacga agcgttccac
     3661 aaggtccagg atgccgtcaa caacaacgcc caagccctct ccaaactcgc ctcagagctt
     3721 tccaatacct tcggcgccat ttccgcctcc atcggcgaca ttatccagag gttggaccct
     3781 cctgaacaag atgcgcagat cgaccgcctc atcaacggca ggcttaccac cctgaacgcc
     3841 ttcgtggctc agcagctcgt gcggagcgag agtgctgccc tgtcggctca gttggcaaag
     3901 gacaaagtga acgaatgcgt gaaggcccag tcaaagcggt cgggcttctg cggtcaagga
     3961 acccacattg tgtccttcgt cgtgaacgcc ccgaatggcc tgtacttcat gcatgtcggc
     4021 tactacccct ccaaccacat cgaagtcgtg tccgcgtacg gactctgcga tgccgcgaac
     4081 cccaccaact gtattgcccc cgtgaacggc tactttatta agaccaacaa cactcggatc
     4141 gtggatgagt ggtcgtacac cgggagctcc ttctacgctc cggaacctat cacgtcactg
     4201 aacaccaaat acgtcgcgcc gcaagtcacc tatcagaaca tctcgactaa cttgcctccc
     4261 cctctgctgg gaaacagcac cggcattgac tttcaagacg agctggacga attcttcaag
     4321 aacgtcagca cctccatacc caacttcgga tcccttaccc aaatcaatac caccttgctc
     4381 gacctcacct acgagatgct gtcactgcaa caggtcgtca aggccctgaa cgaaagttac
     4441 attgacctca aggagctggg gaactacacc tacggaagcg gatacatccc tgaggcacca
     4501 agggacgggc aggcatacgt ccggaaggac ggagaatggg tgctgctgtc caccttcctg
     4561 ggacggagcc tggaagtgct gttccaaggc ccgggacatc accaccatca ccatcatcac
     4621 tcagcatggt cgcaccccca atttgagaag ggcggtggtt ccggtggagg aggatcggga
     4681 ggttccgcat ggtcccatcc ccaattcgag aaataatcaa ctttcttgta caaagttgtg
     4741 aaattctgca gatatccatc acactggcgg ccgctcgagc atgcatctag agggccctat
     4801 tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg
     4861 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     4921 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     4981 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     5041 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     5101 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     5161 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     5221 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     5281 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     5341 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     5401 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     5461 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     5521 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     5581 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     5641 aggtgtggaa agtccctagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     5701 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     5761 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     5821 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     5881 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     5941 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     6001 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     6061 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     6121 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     6181 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     6241 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     6301 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     6361 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     6421 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     6481 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     6541 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     6601 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     6661 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     6721 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     6781 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     6841 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     6901 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     6961 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     7021 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     7081 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     7141 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     7201 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     7261 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     7321 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     7381 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     7441 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     7501 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     7561 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     7621 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     7681 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     7741 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     7801 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     7861 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     7921 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     7981 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     8041 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     8101 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     8161 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     8221 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     8281 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     8341 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     8401 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     8461 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     8521 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     8581 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     8641 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     8701 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     8761 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     8821 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     8881 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     8941 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     9001 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     9061 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     9121 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     9181 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     9241 ctgacgtcga cggatcggga gatccgcctt tatgtgtaac tcttggctga agctcttaca
     9301 ccaatgctgg gggacatgta cctcccaggg gcccaggaag actacgggag gctacaccaa
     9361 cgtcaatcag aggggcctgt gtagctaccg ataagcggac cctcaagagg gcattagcaa
     9421 tagtgtttat aaggccccct tgttaaccct aaacgggtag catatgcttc ccgggtagta
     9481 gtatatacta tccagactaa ccctaattca atagcatatg ttacccaacg ggaagcatat
     9541 gctatcgaat tagggttagt aaaagggtcc taaggaacag cgatatctcc caccccatga
     9601 gctgtcacgg ttttatttac atggggtcag gattccacga gggtagtgaa ccattttagt
     9661 cacaagggca gtggctgaag atcaaggagc gggcagtgaa ctctcctgaa tcttcgcctg
     9721 cttcttcatt ctccttcgtt tagctaatag aataactgct gagttgtgaa cagtaaggtg
     9781 tatgtgaggt gctcgaaaac aaggtttcag gtgacgcccc cagaataaaa tttggacggg
     9841 gggttcagtg gtggcattgt gctatgacac caatataacc ctcacaaacc ccttgggcaa
     9901 taaatactag tgtaggaatg aaacattctg aatatcttta acaatagaaa tccatggggt
     9961 ggggacaagc cgtaaagact ggatgtccat ctcacacgaa tttatggcta tgggcaacac
    10021 ataatcctag tgcaatatga tactggggtt attaagatgt gtcccaggca gggaccaaga
    10081 caggtgaacc atgttgttac actctatttg taacaagggg aaagagagtg gacgccgaca
    10141 gcagcggact ccactggttg tctctaacac ccccgaaaat taaacggggc tccacgccaa
    10201 tggggcccat aaacaaagac aagtggccac tctttttttt gaaattgtgg agtgggggca
    10261 cgcgtcagcc cccacacgcc gccctgcggt tttggactgt aaaataaggg tgtaataact
    10321 tggctgattg taaccccgct aaccactgcg gtcaaaccac ttgcccacaa aaccactaat
    10381 ggcaccccgg ggaatacctg cataagtagg tgggcgggcc aagatagggg cgcgattgct
    10441 gcgatctgga ggacaaatta cacacacttg cgcctgagcg ccaagcacag ggttgttggt
    10501 cctcatattc acgaggtcgc tgagagcacg gtgggctaat gttgccatgg gtagcatata
    10561 ctacccaaat atctggatag catatgctat cctaatctat atctgggtag cataggctat
    10621 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
    10681 cctaatttat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
    10741 cctaatctat atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat
    10801 cctaatagag attagggtag tatatgctat cctaatttat atctgggtag catatactac
    10861 ccaaatatct ggatagcata tgctatccta atctatatct gggtagcata tgctatccta
    10921 atctatatct gggtagcata ggctatccta atctatatct gggtagcata tgctatccta
    10981 atctatatct gggtagtata tgctatccta atttatatct gggtagcata ggctatccta
    11041 atctatatct gggtagcata tgctatccta atctatatct gggtagtata tgctatccta
    11101 atctgtatcc gggtagcata tgctatcctc atgcatatac agtcagcata tgatacccag
    11161 tagtagagtg ggagtgctat cctttgcata tgccgccacc tcccaagggg gcgtgaattt
    11221 tcgctgcttg tccttttcct gctggggatc tcccgatccc ctatggtgca ctctcagtac
    11281 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
    11341 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
    11401 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgatcacaac tttgtataga
    11461 aaagttggcc ctgcagg
//
