LOCUS       sequence-327089-       11465 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of HCoV-OC43 soluble spike trimer protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11465)
  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 11465)
  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: R619-M66-303: CMV51p> HCoV-OC43
            S-2P-T4f-3C-Hi...
FEATURES             Location/Qualifiers
     source          1..11465
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      204..1993
                     /label=oriP
                     /note="Epstein-Barr virus oriP replication origin (Yates et
                     al., 2000)"
     primer_bind     complement(2030..2049)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    2199..2219
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     enhancer        2230..2533
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2534..2737
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2688..2708
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2734..2758
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2803..2827
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             6787..6810
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             6814..6837
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             6844..6867
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             6907..6930
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    complement(6932..6956)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(7018..7036)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(7019..7036)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(7056..7073)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7062..7286
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7332..7760
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7419..7438)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7629..7650
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7769..7789)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7774..8103
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7954..8089
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8016..8035
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             8170..8964
                     /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(8224..8243)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8834..8853
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    9138..9259
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9175..9194)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9229..9248
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9308..9324)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9308..9324)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9321..9343)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9332..9348
                     /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(9356..9362,9363..9380,9381..9386))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9401..9422
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9539..9556)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9710..10298)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9790..9809)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10469..11260,11261..11329))
                     /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     11092..11111
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11330..11434)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatccgcc tttatgtgta actcttggct gaagctctta caccaatgct
       61 gggggacatg tacctcccag gggcccagga agactacggg aggctacacc aacgtcaatc
      121 agaggggcct gtgtagctac cgataagcgg accctcaaga gggcattagc aatagtgttt
      181 ataaggcccc cttgttaacc ctaaacgggt agcatatgct tcccgggtag tagtatatac
      241 tatccagact aaccctaatt caatagcata tgttacccaa cgggaagcat atgctatcga
      301 attagggtta gtaaaagggt cctaaggaac agcgatatct cccaccccat gagctgtcac
      361 ggttttattt acatggggtc aggattccac gagggtagtg aaccatttta gtcacaaggg
      421 cagtggctga agatcaagga gcgggcagtg aactctcctg aatcttcgcc tgcttcttca
      481 ttctccttcg tttagctaat agaataactg ctgagttgtg aacagtaagg tgtatgtgag
      541 gtgctcgaaa acaaggtttc aggtgacgcc cccagaataa aatttggacg gggggttcag
      601 tggtggcatt gtgctatgac accaatataa ccctcacaaa ccccttgggc aataaatact
      661 agtgtaggaa tgaaacattc tgaatatctt taacaataga aatccatggg gtggggacaa
      721 gccgtaaaga ctggatgtcc atctcacacg aatttatggc tatgggcaac acataatcct
      781 agtgcaatat gatactgggg ttattaagat gtgtcccagg cagggaccaa gacaggtgaa
      841 ccatgttgtt acactctatt tgtaacaagg ggaaagagag tggacgccga cagcagcgga
      901 ctccactggt tgtctctaac acccccgaaa attaaacggg gctccacgcc aatggggccc
      961 ataaacaaag acaagtggcc actctttttt ttgaaattgt ggagtggggg cacgcgtcag
     1021 cccccacacg ccgccctgcg gttttggact gtaaaataag ggtgtaataa cttggctgat
     1081 tgtaaccccg ctaaccactg cggtcaaacc acttgcccac aaaaccacta atggcacccc
     1141 ggggaatacc tgcataagta ggtgggcggg ccaagatagg ggcgcgattg ctgcgatctg
     1201 gaggacaaat tacacacact tgcgcctgag cgccaagcac agggttgttg gtcctcatat
     1261 tcacgaggtc gctgagagca cggtgggcta atgttgccat gggtagcata tactacccaa
     1321 atatctggat agcatatgct atcctaatct atatctgggt agcataggct atcctaatct
     1381 atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct atcctaattt
     1441 atatctgggt agcataggct atcctaatct atatctgggt agcatatgct atcctaatct
     1501 atatctgggt agtatatgct atcctaatct gtatccgggt agcatatgct atcctaatag
     1561 agattagggt agtatatgct atcctaattt atatctgggt agcatatact acccaaatat
     1621 ctggatagca tatgctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     1681 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     1741 ctgggtagta tatgctatcc taatttatat ctgggtagca taggctatcc taatctatat
     1801 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatctgtat
     1861 ccgggtagca tatgctatcc tcatgcatat acagtcagca tatgataccc agtagtagag
     1921 tgggagtgct atcctttgca tatgccgcca cctcccaagg gggcgtgaat tttcgctgct
     1981 tgtccttttc ctgctgggga tctcccgatc ccctatggtg cactctcagt acaatctgct
     2041 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     2101 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     2161 atctgcttag ggttaggcgt tttgcgctgc ttcgatcaca actttgtata gaaaagttgg
     2221 ccctgcaggc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     2281 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     2341 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     2401 tcatatgcca agtccgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     2461 tgcccagtac atgaccttac gggactttcc tacttggcag tacatctacg tattagtcat
     2521 cgctattacc atggtgatgc ggttttggca gtacaccaat gggcgtggat agcggtttga
     2581 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     2641 aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg
     2701 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc
     2761 ctggagacgc catccacgct gttttgacct ccatagaaga caacaacttt gtacaaaaaa
     2821 gttggcacca tgttcctgat tctgttaatt tcgctgccga ctgcgtttgc ggtcatcggg
     2881 gacctgaagt gtcccctcga ctcccggacc ggtagcctga acaatattga taccggccct
     2941 ccgtccatct ccaccgcaac cgtcgatgtg accaacggac tcggtaccta ctacgtcctg
     3001 gacagagtct atctgaacac caccctgttc cttaacgggt actaccctac ctccggatcg
     3061 acctaccgga acatggccct gaagggaacc gacaagctgt caaccctgtg gttcaagccg
     3121 ccgtttctta gtgacttcat caacgggatt tttgcgaagg tcaagaacac caaggtgttc
     3181 aaggacggag tgatgtactc cgagttcccc gccattacca ttgggtccac gtttgtcaac
     3241 accagctatt ccgtggttgt gcagcctcgg actatcaaca gcacccagga cggcgtgaat
     3301 aagctgcagg gactcctgga ggtgtccgtg tgtcagtaca acatgtgtga atacccccac
     3361 actatttgcc accccaagct cgggaaccac ttcaaggaac tgtggcacat ggacactggg
     3421 gtggtgtcct gcctgtacaa gcgcaacttc acttacgacg tgaacgccac ttacctgtac
     3481 tttcacttct atcaagaggg aggaaccttc tacgcctact ttactgacac cggcgtggtg
     3541 accaagttcc tgttcaacgt gtacctgggg atggcactgt cccactacta cgtgatgccc
     3601 ctgacgtgca tctcccggcg ggatatcggc ttcactctcg agtactgggt gacccctctg
     3661 acctcacgcc agtacctcct ggccttcaac caggacggca ttatctttaa cgccgtcgat
     3721 tgcatgtctg acttcatgag cgagatcaag tgtaaaaccc agtcgattgc accaccaacc
     3781 ggagtgtacg aactgaacgg atacacggtg caaccaatcg cggatgtgta caggcggaag
     3841 ccagacctcc ccaactgcaa catcgaagcc tggctgaacg acaagtcggt cccttcaccg
     3901 ctcaactggg agcgaaaaac cttctccaat tgcaacttta acatgtcgtc actgatgagc
     3961 ttcatccagg cggactcatt cacttgcaac aacatcgacg ccgccaaaat ctacgggatg
     4021 tgtttctcct ccatcaccat tgacaagttc gccattccta acgggaggaa ggtggacctc
     4081 cagcttggca acctcggcta tctgcagtcc ttcaactacc ggatcgacac caccgccacg
     4141 tcctgccagc tctactataa cctccccgcc gctaacgtgt ccgtgagccg gttcaaccct
     4201 tcaacctgga ataagcgctt cggcttcatc gaaaactcgg tgttcaagcc tcagcctgct
     4261 ggagtgctga ctaatcacga cgtggtgtac gcccagcact gcttcaaggc cccgaagaac
     4321 ttctgcccgt gcaaattgaa ctcgtcgctg tgcgtgggct caggacccgg aaagaacaac
     4381 ggcatcggaa cttgccctgc gggaaccaac tacctgactt gccacaacct gtgcaaccct
     4441 gaccccataa ctttcaccgg tccgtacaag tgtccgcaaa ccaagtccct cgtgggaatt
     4501 ggagagcact gcagcggcct tgccgtgaag tccgattact gcggaggaaa cccctgcaca
     4561 tgccagccgc aggccttcct cggctggtcg gctgacagct gcctgcaggg cgataagtgt
     4621 aacatcttcg ccaacctcat cctgcatgac gtgaactcgg gactcacctg ttccaccgac
     4681 ctccagaagg ctaacaccga tatcaagttg ggagtctgtg tcaactatga cctgtacgga
     4741 atcagcggcc agggcatctt tgtggaggtc aacgccacct actacaactc ctggcagaat
     4801 ctgctgtacg actccaacgg caatctgtac gggttcagag actacattac taatcgcact
     4861 tttatgatcc gctcctgcta ctccggacgc gtgtcagccg cattccatgc caactcctcg
     4921 gaacccgccc tgctgtttcg caacatcaag tgcaactacg tgttcaacaa cagcctcatt
     4981 cggcaactgc agccgatcaa ctactttgat tcgtacctgg gatgtgtcgt gaacgcctac
     5041 aactccaccg ccatctccgt ccaaacttgc gatcttaccg tgggctccgg ctactgcgtg
     5101 gactactcaa agaacggtgg cagcggcaga gccattacca ccggataccg cttcaccaac
     5161 ttcgagcctt tcactgtgaa ctccgtgaat gactccctgg aacccgtcgg cggtctgtac
     5221 gagatccaga tcccgtccga attcacaatc ggcaacatgg aagaattcat ccaaacctcc
     5281 agccccaaag tcactatcga ttgcgccgca ttcgtgtgcg gggactacgc agcgtgcaaa
     5341 tcccaactgg tggaatacgg atccttctgc gacaacatta acgcgatcct gaccgaagtc
     5401 aacgaactgc tggacactac ccagcttcaa gtggccaatt ccctgatgaa cggcgtgact
     5461 ctgagcacca agctgaagga tggcgtcaac ttcaacgtgg atgacataaa cttcagctcc
     5521 gtgctggggt gtttgggttc tgagtgctcc aaggcctcat cccgctccgc cattgaggac
     5581 ctcctgttcg acaaggtcaa gctctccgac gtgggctttg tggccgccta caacaactgt
     5641 accggaggcg ccgagatcag agatttgatc tgcgtccagt cctacaaggg tatcaaggtg
     5701 ctgccgccac tcctcagcga gaaccagatc agcgggtaca ccctcgctgc tacctccgca
     5761 tcgctgttcc cgccgtggac tgcagctgcc ggagtgccat tctacctaaa tgtgcagtac
     5821 cggattaacg gtctgggagt cactatggac gtgctgagcc agaaccagaa gctcatcgcc
     5881 aacgcgttca acaatgcgct ggacgcgatc caggaaggct ttgatgcaac caacagcgca
     5941 ctcgtgaaga tccaggccgt agtgaacgcg aacgctgagg ccctgaacaa cttgctgcag
     6001 cagttgtcaa acagattcgg agccatctcc tcgagcctgc aggaaatcct gtcccggttg
     6061 gatcctcctg aagccgaggc tcaaatcgac cggctcatta acgggaggct gactgccttg
     6121 aacgcttacg tcagccagca attgtccgac tccacgctcg tgaagttctc cgccgcccaa
     6181 gctatggaaa aggtcaacga atgcgtgaag tctcaatcct caagaatcaa cttttgcgga
     6241 aacggcaacc acatcatttc gctggtgcag aacgccccct atggcctgta cttcatccat
     6301 ttctcctacg tgccaaccaa atacgtgact gccaaagtgt cgccgggtct gtgtattgcc
     6361 ggggacaggg gaatcgcccc taagtccgga tacttcgtga acgtgaacaa cacttggatg
     6421 tacaccggca gcggatacta ctaccctgag cccattactg aaaacaacgt cgtcgtgatg
     6481 tccacctgtg ctgtgaacta cactaaggcc ccttacgtga tgctgaacac atccacccct
     6541 aatctgccgg atttcagaga agaactggac cagtggttca agaaccagac cagcgtggcg
     6601 ccagacctct cacttgacta catcaatgtg accttcttgg atctgcaagt ggagatgaat
     6661 cggctccagg aagcgattaa ggtcctgaac gggtctggct atatccccga agccccaagg
     6721 gatggacagg catacgtgcg caaggatgga gaatgggtgc ttttgtcaac ctttttgggt
     6781 cggtccctgg aagtgctgtt ccaagggccg ggtcatcacc accatcacca tcatcactcc
     6841 gcttggagcc accctcagtt cgaaaaggga ggaggatccg gcggtggagg atcgggagga
     6901 tccgcctggt cgcatcccca gttcgagaag taatcaactt tcttgtacaa agttgtgaaa
     6961 ttctgcagat atccatcaca ctggcggccg ctcgagcatg catctagagg gccctattct
     7021 atagtgtcac ctaaatgcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca
     7081 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     7141 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     7201 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     7261 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     7321 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     7381 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     7441 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     7501 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     7561 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     7621 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     7681 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     7741 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     7801 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     7861 tgtggaaagt ccctaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     7921 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     7981 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     8041 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     8101 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     8161 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     8221 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     8281 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     8341 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     8401 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     8461 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     8521 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     8581 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     8641 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     8701 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     8761 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     8821 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     8881 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     8941 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     9001 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     9061 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     9121 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     9181 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     9241 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     9301 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     9361 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     9421 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     9481 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     9541 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     9601 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     9661 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     9721 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     9781 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     9841 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     9901 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     9961 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
    10021 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
    10081 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
    10141 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
    10201 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
    10261 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
    10321 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
    10381 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
    10441 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
    10501 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
    10561 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
    10621 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
    10681 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
    10741 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
    10801 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
    10861 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
    10921 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
    10981 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
    11041 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
    11101 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
    11161 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
    11221 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
    11281 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
    11341 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
    11401 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
    11461 acgtc
//
