LOCUS       sequence-334256-       11219 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of SARS-CoV-2 soluble spike trimer protein 
            (B.1.1.7 variant).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11219)
  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 11219)
  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: "Protein 
            Expr Purif."; date: "2020-06-4"; volume: "174:105686. doi"; pages: "
            10.1016/j.pep.2020.105686"
COMMENT     Sequence Label: R620-M01-303: CMV51p> SARS-CoV-2
            S(1-1208)-2P-...
FEATURES             Location/Qualifiers
     source          1..11219
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(325..344)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    494..514
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     enhancer        525..828
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        829..1032
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     983..1003
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1029..1053
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    1098..1122
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4836..4859
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4863..4886
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             4893..4916
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4956..4979
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    complement(4981..5005)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(5067..5085)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5068..5085)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5105..5122)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5111..5335
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5381..5809
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5468..5487)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5678..5699
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5818..5838)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5823..6152
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6003..6138
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6065..6084
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6219..7013
                     /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(6273..6292)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6883..6902
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7187..7308
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7224..7243)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7278..7297
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7357..7373)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7357..7373)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7370..7392)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7381..7397
                     /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(7405..7411,7412..7429,7430..7435))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7450..7471
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7588..7605)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7759..8347)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7839..7858)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8518..9309,9310..9378))
                     /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     9141..9160
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9379..9483)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(9718..11219,1..288)
                     /label=oriP
                     /note="Epstein-Barr virus oriP replication origin (Yates et
                     al., 2000)"
ORIGIN
        1 tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatt
       61 tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc
      121 tatatctggg tagtatatgc tatcctaatc tgtatccggg tagcatatgc tatcctcatg
      181 catatacagt cagcatatga tacccagtag tagagtggga gtgctatcct ttgcatatgc
      241 cgccacctcc caagggggcg tgaattttcg ctgcttgtcc ttttcctgct ggggatctcc
      301 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
      361 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
      421 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
      481 gctgcttcga tcacaacttt gtatagaaaa gttggccctg caggcgttac ataacttacg
      541 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      601 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      661 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtcc gccccctatt
      721 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttacgggac
      781 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      841 tggcagtaca ccaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      901 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      961 cgtaataacc ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     1021 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
     1081 gacctccata gaagacaaca actttgtaca aaaaagttgg caccatgttc gtgttccttg
     1141 tcctcctgcc gctggtgtca tcgcagtgcg tgaacctgac cacccggact cagctgcctc
     1201 ctgcttacac taactccttc actcgcgggg tgtactaccc tgacaaagtg ttccggtcgt
     1261 ccgtgctgca tagcacccag gacctgttcc ttccgttctt ctccaacgtg acctggttcc
     1321 acgccatttc ggggaccaac ggcactaaga gattcgacaa ccccgtcttg cccttcaatg
     1381 acggagtata cttcgcctcg acggaaaagt ccaacatcat tcggggttgg atcttcggca
     1441 ctaccctgga cagcaagacc cagtccctgc tcattgtcaa caacgccacc aacgtcgtga
     1501 tcaaagtctg cgagtttcag ttttgtaacg atccgttcct cggcgtgtac cacaagaaca
     1561 acaagagctg gatggagtcc gaattccggg tctactcctc cgccaacaac tgtaccttcg
     1621 aatacgtgtc ccaacctttc ctcatggacc tcgaaggaaa acagggaaac ttcaagaact
     1681 tgagagagtt tgtgttcaag aatatcgatg gctacttcaa gatctactcg aagcataccc
     1741 cgatcaacct cgtgcgggac ctacctcaag ggttctccgc acttgagccg ctcgtggacc
     1801 tccccattgg catcaacatc accagattcc agactttact ggcactgcat agatcgtatc
     1861 tcacacccgg ggactcctcc tccgggtgga cagctggtgc tgcggcctac tacgtgggat
     1921 atctccagcc ccggaccttc ttgctgaagt acaacgagaa cggcaccatc acggatgccg
     1981 tggactgtgc actggacccc ttgagtgaaa ccaagtgcac gctgaagtcc ttcaccgtgg
     2041 aaaaaggcat ctaccagacc agcaacttca gggtgcagcc tactgaatca atcgtgcggt
     2101 tccctaacat taccaatctg tgcccgttcg gcgaagtgtt caacgccact agattcgcct
     2161 cggtgtacgc ctggaaccga aagcggatca gcaactgcgt ggccgactac tcagtgctct
     2221 acaactcggc cagcttctcc accttcaagt gctacggagt gtcgcccacc aagctgaacg
     2281 atctgtgctt caccaacgtg tacgcggatt ccttcgtgat ccgcggagat gaagtccgcc
     2341 aaatcgcccc cggtcaaacc ggaaagattg ccgactacaa ctataagctg ccggacgact
     2401 ttaccggatg cgtgattgcc tggaattcca acaatttgga ttcaaaagtc ggcggcaact
     2461 acaactacct gtaccgcctg ttccggaagt caaacctgaa gccgttcgaa cgcgacatca
     2521 gcaccgaaat ctatcaagcg ggaagcactc cctgcaacgg ggtggagggc ttcaactgct
     2581 acttcccgct gcaaagctac ggattccagc ctacctacgg cgttggatac cagccatatc
     2641 gcgtggtcgt gctctccttt gaacttctgc acgcccctgc cactgtgtgc ggccccaaga
     2701 agtccactaa cctcgtcaag aataagtgcg tgaatttcaa ctttaacgga ttgaccggga
     2761 ccggagtgct caccgagtcc aacaagaaat tcctgccgtt ccaacagttt ggacgggaca
     2821 tcgatgatac tacggatgct gtgcgggacc cacagactct ggagatcctg gacattactc
     2881 cctgttcctt tggcggggtg tcggtcatca cccctggtac taacaccagc aaccaggtcg
     2941 ccgtgctgta tcagggggtg aactgcaccg aagtgcccgt ggcgatccac gccgaccagc
     3001 tgactccaac ttggcgcgtg tactcgactg gttcgaacgt gttccagact cgggccggtt
     3061 gcctgatcgg agcggagcat gtcaacaaca gctacgagtg cgacatccct attggagccg
     3121 ggatttgcgc cagctaccag acccagacca attcgcatgg gtcggccagc agcgtggctt
     3181 cccaatcgat catcgcctac accatgtccc tgggagccga gaactcagtg gcgtactcca
     3241 acaactcgat tgccatcccc atcaatttca cgatcagcgt gaccaccgag atcctgcctg
     3301 tgtccatgac caagacctcc gtggactgca ccatgtacat ctgcggagac tccaccgagt
     3361 gctccaacct cctgctgcaa tacggctcgt tctgtaccca actgaaccga gccctgactg
     3421 ggatcgccgt ggagcaggac aagaacacac aggaggtgtt cgcacaagtc aagcagatct
     3481 acaagacccc gcccatcaag gatttcggag ggttcaattt ctcccaaatt ctgccggacc
     3541 cttccaagcc gtccaagagg tcattcattg aggatctcct gttcaacaag gtcaccctgg
     3601 ccgatgctgg tttcatcaag cagtacggag actgcctggg ggacatcgca gcccgcgatc
     3661 tgatctgcgc ccaaaagttc aacggcctga ccgtcctgcc tccgcttctg actgatgaaa
     3721 tgattgccca gtacacctct gcgctgctgg ctggcactat cacctccggc tggaccttcg
     3781 gcgctggagc agcactgcag attccatttg ccatgcaaat ggcctaccgc ttcaacggaa
     3841 tcggagtgac tcagaacgtc ctgtacgaga accagaagct catcgccaac cagttcaact
     3901 ccgccattgg gaagattcag gattccctgt cgagcactgc gtcagccctg ggaaagctgc
     3961 aggacgtcgt gaaccagaac gcacaggccc tgaacaccct tgtgaagcag ctgtcgtcga
     4021 acttcggggc gatctcctcc gtgttgaacg acatcctggc ccggctggac ccgcctgagg
     4081 cagaagtgca gatcgaccgg ctgattaccg gacgcctcca gtcccttcaa acttatgtga
     4141 cccagcagct gattcgcgcg gcggaaattc gggctagcgc caatctggct gccaccaaga
     4201 tgagcgaatg cgtgcttgga cagtccaaga gagtggactt ttgtggaaag ggctaccacc
     4261 tgatgagctt cccccaatcc gccccacacg gtgtcgtgtt tctgcacgtg acctacgtgc
     4321 cggcgcagga aaagaacttc accactgccc ctgccatctg ccacgatggg aaggcgcact
     4381 tcccgaggga gggagtgttc gtgtcaaacg gaacccactg gttcgtgacc caaaggaact
     4441 tctacgagcc ccagattatc accacccaca acactttcgt gagcggcaac tgcgacgtgg
     4501 tcatcggcat cgtgaacaac actgtgtacg acccgctgca accggagctc gatagcttta
     4561 aggaagaact cgacaagtac tttaagaatc atacgtcccc cgatgtggat ctgggcgaca
     4621 ttagcggtat caacgcgtct gtcgtgaaca ttcagaagga gattgacagg ctcaacgaag
     4681 tcgccaaaaa cctgaacgag agcctgatag acttgcagga actgggaaaa tacgagcagg
     4741 gctccggtta catccccgaa gctccacggg acggacaggc ctacgtgcgc aaggacggag
     4801 aatgggtgct gctgagcacc ttcctcgggc ggtccctgga ggtgctgttc caaggcccag
     4861 gccaccatca tcatcaccac caccactcgg cttggtcaca cccccaattc gaaaagggcg
     4921 gtggttctgg aggaggcgga tccggaggat ccgcgtggag tcatccgcag tttgaaaagt
     4981 aatcaacttt cttgtacaaa gttgtgaaat tctgcagata tccatcacac tggcggccgc
     5041 tcgagcatgc atctagaggg ccctattcta tagtgtcacc taaatgctag agctcgctga
     5101 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     5161 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     5221 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     5281 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     5341 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     5401 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     5461 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     5521 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     5581 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     5641 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     5701 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     5761 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     5821 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5881 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cctaggctcc ccagcaggca
     5941 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     6001 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     6061 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     6121 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     6181 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     6241 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     6301 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     6361 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     6421 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     6481 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     6541 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     6601 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     6661 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     6721 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     6781 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     6841 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     6901 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     6961 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     7021 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     7081 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     7141 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     7201 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     7261 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     7321 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     7381 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     7441 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     7501 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     7561 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     7621 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     7681 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     7741 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     7801 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     7861 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     7921 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     7981 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     8041 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     8101 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     8161 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     8221 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     8281 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     8341 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     8401 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     8461 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     8521 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     8581 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     8641 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     8701 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     8761 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     8821 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     8881 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     8941 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     9001 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     9061 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     9121 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     9181 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     9241 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     9301 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     9361 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     9421 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     9481 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc cgcctttatg
     9541 tgtaactctt ggctgaagct cttacaccaa tgctggggga catgtacctc ccaggggccc
     9601 aggaagacta cgggaggcta caccaacgtc aatcagaggg gcctgtgtag ctaccgataa
     9661 gcggaccctc aagagggcat tagcaatagt gtttataagg cccccttgtt aaccctaaac
     9721 gggtagcata tgcttcccgg gtagtagtat atactatcca gactaaccct aattcaatag
     9781 catatgttac ccaacgggaa gcatatgcta tcgaattagg gttagtaaaa gggtcctaag
     9841 gaacagcgat atctcccacc ccatgagctg tcacggtttt atttacatgg ggtcaggatt
     9901 ccacgagggt agtgaaccat tttagtcaca agggcagtgg ctgaagatca aggagcgggc
     9961 agtgaactct cctgaatctt cgcctgcttc ttcattctcc ttcgtttagc taatagaata
    10021 actgctgagt tgtgaacagt aaggtgtatg tgaggtgctc gaaaacaagg tttcaggtga
    10081 cgcccccaga ataaaatttg gacggggggt tcagtggtgg cattgtgcta tgacaccaat
    10141 ataaccctca caaacccctt gggcaataaa tactagtgta ggaatgaaac attctgaata
    10201 tctttaacaa tagaaatcca tggggtgggg acaagccgta aagactggat gtccatctca
    10261 cacgaattta tggctatggg caacacataa tcctagtgca atatgatact ggggttatta
    10321 agatgtgtcc caggcaggga ccaagacagg tgaaccatgt tgttacactc tatttgtaac
    10381 aaggggaaag agagtggacg ccgacagcag cggactccac tggttgtctc taacaccccc
    10441 gaaaattaaa cggggctcca cgccaatggg gcccataaac aaagacaagt ggccactctt
    10501 ttttttgaaa ttgtggagtg ggggcacgcg tcagccccca cacgccgccc tgcggttttg
    10561 gactgtaaaa taagggtgta ataacttggc tgattgtaac cccgctaacc actgcggtca
    10621 aaccacttgc ccacaaaacc actaatggca ccccggggaa tacctgcata agtaggtggg
    10681 cgggccaaga taggggcgcg attgctgcga tctggaggac aaattacaca cacttgcgcc
    10741 tgagcgccaa gcacagggtt gttggtcctc atattcacga ggtcgctgag agcacggtgg
    10801 gctaatgttg ccatgggtag catatactac ccaaatatct ggatagcata tgctatccta
    10861 atctatatct gggtagcata ggctatccta atctatatct gggtagcata tgctatccta
    10921 atctatatct gggtagtata tgctatccta atttatatct gggtagcata ggctatccta
    10981 atctatatct gggtagcata tgctatccta atctatatct gggtagtata tgctatccta
    11041 atctgtatcc gggtagcata tgctatccta atagagatta gggtagtata tgctatccta
    11101 atttatatct gggtagcata tactacccaa atatctggat agcatatgct atcctaatct
    11161 atatctgggt agcatatgct atcctaatct atatctgggt agcataggct atcctaatc
//
