LOCUS       sequence-327090-       11477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of HCoV-NL63 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 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-M91-303: CMV51p> HCoV-NL63
            S-2P-T4f-3C-Hi...
FEATURES             Location/Qualifiers
     source          1..11477
                     /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             complement(3193..3204)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             6799..6822
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             6826..6849
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             6856..6879
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             6919..6942
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    complement(6944..6968)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(7030..7048)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(7031..7048)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(7068..7085)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7074..7298
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7344..7772
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7431..7450)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7641..7662
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7781..7801)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7786..8115
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7966..8101
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8028..8047
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             8182..8976
                     /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(8236..8255)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8846..8865
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    9150..9271
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9187..9206)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9241..9260
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9320..9336)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9320..9336)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9333..9355)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9344..9360
                     /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(9368..9374,9375..9392,9393..9398))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9413..9434
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9551..9568)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9722..10310)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9802..9821)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10481..11272,11273..11341))
                     /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     11104..11123
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11342..11446)
                     /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 tgaagctgtt cctgatcctg ctggtgctgc cgctcgcttc ctgcttcttc
     2881 acctgtaaca gcaacgcgaa cctgtccatg ctgcaactcg gggtgcccga taactcctca
     2941 actatcgtga ccggactgtt acccacccac tggttctgcg caaatcagtc gacctccgtg
     3001 tacagcgcga acgggttctt ctacattgat gtcggaaacc atagatccgc cttcgcactg
     3061 cacaccgggt actacgacgc caaccagtat tacatctacg tgaccaacga aatcggtctg
     3121 aatgcctcag tgaccctgaa gatctgcaaa ttctcccgca atactacttt tgattttctg
     3181 agcaacgcat cctcgtcctt cgattgcatc gtgaaccttc tgttcaccga gcaattgggt
     3241 gccccgcttg gaattaccat ctccggagag actgtccggc tgcacctgta caacgtcact
     3301 cgcaccttct acgtgcccgc cgcttacaag ctcactaagc tgagcgtcaa gtgctacttc
     3361 aactactcct gcgtgttctc ggtggtcaac gcaactgtga cggttaacgt caccacccac
     3421 aacggccgcg tggtcaacta cactgtgtgc gacgactgca atggatacac agacaacatt
     3481 ttctcggtgc aacaggacgg acgcattccg aacggattcc ctttcaacaa ctggttcttg
     3541 ctgaccaacg gcagcaccct ggtcgacgga gtgtcgcggc tgtaccagcc actgcgcctg
     3601 acttgcttgt ggccagtgcc tgggctcaag tccagcacag gctttgtgta ctttaacgct
     3661 accggatccg acgtgaactg taacggctac cagcacaact cggtggtgga tgtgatgcgg
     3721 tacaacctta acttctccgc taattccctc gacaacctga agtcgggcgt gatcgtgttt
     3781 aagaccctcc agtacgacgt gctgttctac tgctccaact cgagctccgg agtgctcgac
     3841 acaaccattc ccttcggacc atcaagccag ccctactact gctttatcaa cagcaccatc
     3901 aacaccactc atgtgtccac tttcgtggga atcctgccac cgaccgtcag ggaaatcgtg
     3961 gtggccagga ctggccagtt ctacatcaac ggcttcaagt acttcgacct cggattcatt
     4021 gaggccgtga acttcaatgt cactaccgcg tccgcgactg acttctggac tgtggccttc
     4081 gccaccttcg tggacgtgct ggtcaacgtg tcggccacta acatccagaa cctcctgtac
     4141 tgcgactcac cgttcgagaa gctgcagtgc gagcatctgc agtttggcct gcaagatgga
     4201 ttctactccg ccaacttcct cgacgacaac gtgctccccg aaacctatgt ggccttgcct
     4261 atctattacc aacacaccga catcaacttt accgccaccg cgtcgttcgg aggatcctgc
     4321 tacgtctgca agccccacca ggtcaacatc tccttgaacg gcaatacctc agtgtgcgtg
     4381 cgcacctccc acttctcaat tcggtacatc tacaatagag tgaagtctgg ttcacctggg
     4441 gacagctcat ggcatatcta cctgaagtcc ggcacctgtc cgtttagctt cagcaagctg
     4501 aacaactttc agaagttcaa aaccatctgc ttctccaccg tcgaggtgcc agggagctgt
     4561 aacttcccgc tcgaagccac ctggcactac acctcataca ccatcgtggg agccctttac
     4621 gtgacctggt ctgaaggaaa cagcatcact ggagtgccgt acccagtgtc cggcatcagg
     4681 gagttctcca acctggtgct caacaactgc accaagtaca acatctacga ctacgttggc
     4741 actggcatca tcagatccag caaccagtca ctcgccggtg gcatcaccta cgtgtccaac
     4801 tcgggaaacc tcctgggatt caagaacgtg tccaccggca acatcttcat cgtgaccccc
     4861 tgcaaccagc cggatcaagt cgcggtgtac cagcagtcca tcatcggagc catgacggcc
     4921 gtgaacgaat caagatacgg gttgcaaaat ctcctgcaac tgccgaactt ctactacgtg
     4981 tcgaacggag gaaacaattg cacgactgcc gtcatgacct acagcaactt cggaatttgt
     5041 gccgatgggt ccctgatccc ggtccggcct agaaactcct cggacaacgg catcagcgcg
     5101 atcatcaccg ccaatctgtc cattccatca aactggacca ctagcgtcca ggtcgagtat
     5161 ctgcagatca ccagcacccc gattgtggtg gattgcgcca cttacgtgtg caacgggaat
     5221 ccccgctgca agaacctcct gaagcagtat acttccgcct gcaagactat cgaggatgcc
     5281 ctccgccttt ctgcccactt agaaaccaac gatgtgtcca gcatgctgac cttcgatagc
     5341 aacgctttct cgcttgccaa cgtcacctcc ttcggcgatt acaacctttc ctcggtactg
     5401 ccgcagcgga acattcggtc gtcacggatc gcggggagat cagcgctgga ggacctcctg
     5461 ttctcgaaag tggtcacttc gggactgggg actgtcgacg tggactacaa aagttgcact
     5521 aaggggctct ccattgcgga tcttgcttgc gctcaatact acaacgggat tatggtgctg
     5581 cctggagtgg ccgatgccga aagaatggcc atgtacaccg gctcccttat cggaggaatg
     5641 gtcctcggcg gactcacctc tgccgcggca atcccctttt ccctggcctt gcaagcccgc
     5701 ctgaactacg tggctctgca gactgacgtg ctgcaggaga accagaagat tctggccgcc
     5761 tcattcaaca aggccatcaa caacatcgtg gcctccttct cctccgtgaa tgatgcgatt
     5821 acccagaccg ccgaagccat ccacactgtc acgattgccc tgaacaagat tcaggatgtc
     5881 gtgaatcagc aaggctcagc cctgaatcac ctgacttcgc aattgcggca caacttccaa
     5941 gcgatctcga acagcatcca ggccatctac gacagactgg atcctccgca agccgaccaa
     6001 caggtcgacc ggctgattac aggacgcctc gcagcactca acgccttcgt gagccaggtc
     6061 ttgaacaagt acactgaagt gcggggatca agacgcctgg cgcagcagaa gattaacgag
     6121 tgcgtcaaga gccagtccaa ccgatacggt ttttgcggga acggcaccca catcttctcc
     6181 attgtgaata gcgcaccgga tggcctcctg ttccttcata ccgtgctgct gcctaccgac
     6241 tacaagaacg tgaaggcttg gagcgggatc tgtgtcgacg gcatctacgg ttacgtgctg
     6301 aggcagccga acctggtgtt gtacagcgac aacggagtgt tcagagtgac gtcccggatc
     6361 atgtttcaac caagacttcc tgtgctctcc gacttcgtgc agatatacaa ctgtaacgtg
     6421 accttcgtga acatttcccg cgtggaactc cacaccgtga ttcctgacta cgtggacgtg
     6481 aacaagaccc tgcaggagtt cgcgcagaat ctccctaaat acgtgaagcc caacttcgac
     6541 ctgaccccct ttaacctgac atacctcaac ctgtcctccg aattgaagca gctggaggcc
     6601 aagaccgcct cgctgttcca aaccaccgtg gagctgcagg gcctgatcga ccagattaat
     6661 tccacctacg tcgacctgaa gctgctgaac cgcttcgaga acggctccgg ctacatcccg
     6721 gaagctcccc gagatgggca ggcctacgtc cggaaggacg gagaatgggt cctcctctcc
     6781 accttcctgg gccgctcgct agaagtgctg ttccaagggc cgggacacca tcaccaccac
     6841 catcatcatt cggcatggtc gcaccctcaa ttcgagaagg gcggaggcag tggaggtgga
     6901 ggttccggcg gatccgcttg gtcccacccg caattcgaaa agtaatcaac tttcttgtac
     6961 aaagttgtga aattctgcag atatccatca cactggcggc cgctcgagca tgcatctaga
     7021 gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct cgactgtgcc
     7081 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     7141 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     7201 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     7261 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     7321 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     7381 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     7441 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     7501 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     7561 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     7621 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     7681 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     7741 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     7801 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     7861 tcagcaacca ggtgtggaaa gtccctaggc tccccagcag gcagaagtat gcaaagcatg
     7921 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     7981 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     8041 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     8101 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     8161 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     8221 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     8281 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     8341 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     8401 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     8461 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     8521 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     8581 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     8641 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     8701 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     8761 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     8821 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     8881 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     8941 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     9001 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     9061 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     9121 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     9181 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     9241 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     9301 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     9361 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     9421 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     9481 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     9541 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     9601 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     9661 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     9721 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     9781 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     9841 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     9901 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     9961 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
    10021 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
    10081 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
    10141 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
    10201 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
    10261 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
    10321 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
    10381 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
    10441 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
    10501 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
    10561 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
    10621 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
    10681 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
    10741 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
    10801 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
    10861 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
    10921 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
    10981 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
    11041 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
    11101 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
    11161 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
    11221 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
    11281 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
    11341 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
    11401 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
    11461 aagtgccacc tgacgtc
//
