LOCUS       sequence-327663-       10928 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of HCoV-229E soluble spike trimer protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10928)
  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 10928)
  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-M90-303: CMV51p> HCoV-229E
            S-2P-T4f-3C-Hi...
FEATURES             Location/Qualifiers
     source          1..10928
                     /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             6250..6273
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             6277..6300
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             6307..6330
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             6370..6393
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    complement(6395..6419)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(6481..6499)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6482..6499)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(6519..6536)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6525..6749
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6795..7223
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6882..6901)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7092..7113
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7232..7252)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7237..7566
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7417..7552
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7479..7498
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             7633..8427
                     /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(7687..7706)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8297..8316
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    8601..8722
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8638..8657)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8692..8711
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8771..8787)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8771..8787)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8784..8806)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8795..8811
                     /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(8819..8825,8826..8843,8844..8849))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8864..8885
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9002..9019)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9173..9761)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9253..9272)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9932..10723,10724..10792))
                     /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     10555..10574
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10793..10897)
                     /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 tgttcgtgct gctggtcgcc tacgccttgc ttcacattgc gggatgccag
     2881 actaccaatg gcaccaacac cagccactcg gtttgcaacg gatgcgtggg ccacagcgaa
     2941 aacgtgttcg ctgtggagtc cggaggatac atcccctcca acttcgcatt caacaactgg
     3001 ttcctgctga ccaacacctc cagcgtggtg gatggtgtcg tcagatcgtt ccagccgctc
     3061 ctgctgaact gcctgtggag cgtgtccggc tcccaattca ccaccgggtt cgtgtacttc
     3121 aacggcactg gaagaggggc gtgcaagggc ttttactcaa acgcctcctc cgacgtgatc
     3181 cgctacaaca tcaacttcga agagaaccta cggagaggca ccatcttgtt caagacctcc
     3241 tatggggccg tggtgtttta ctgcaccaac aacacccttg tgtccgggga tgcccatatc
     3301 ccttccggta ctgtgctggg caacttctac tgtttcgtga acaccaccat cgggaacgaa
     3361 actacctccg ccttcgtcgg agctctgcca aagacagtgc gcgagttcgt catctcgcgg
     3421 actggccatt tctatattaa cggatacaga tacttctcac tgggcgatgt tgaagccgtg
     3481 aacttcaacg tgaccaacgc agccaccacc gtgtgcaccg tcgcactggc ttcctacgcc
     3541 gatgtgctcg tgaacgtgtc gcagactgcc atcgccaaca ttatctactg caactccgtg
     3601 atcaacaggc tgcgctgcga ccagctctcg tttgatgtgc cggatggctt ctactcgacc
     3661 tcgccaatcc aaccggtgga actgcccgtg tccatcgtgt cgctcccggt gtaccacaaa
     3721 cataccttca tcgtgctgta cgtgaacttc gaacatagac ggggacccgg aaagtgctac
     3781 aactgtaggc ccgcagtcat caacatcacg ctggccaact tcaacgaaac caaggggccc
     3841 ctctgtgtcg atacctccca ctttaccacc caatttgtgg acaacgtgaa gctcgcacgg
     3901 tggtctgcgt ccatcaatac cggaaactgc cccttctcct tcggaaaggt caacaacttt
     3961 gtgaagtttg gctccgtgtg cttcagcttg aaggatatcc ctggaggttg tgcgatgccg
     4021 atcatggcca acctggtcaa ttcgaagtcg cacaacattg gtagcctcta cgtatcctgg
     4081 agcgatggcg acgtgatcac cggagtgccc aagcctgtcg aaggggtgtc ctccttcatg
     4141 aacgtgaccc tcaacaagtg cactaagtac aacatctacg atgtgtccgg agtcggtgtc
     4201 attcgcatct caaacgacac cttcctgaac ggaatcacct acacttccac ttcggggaat
     4261 ctccttgggt tcaaggacgt cactaatggc acgatctact caatcacccc gtgtaacccg
     4321 ccggaccagc ttgtggtgta ccagcaggcc gtagtcggag ccatgctgtc tgagaacttc
     4381 acctcatacg gattctcaaa cgtggtggag atgccgaagt tcttttacgc ctctaatggc
     4441 acctacaact gcaccgacgc ggtgctgaca tactcgtcct tcggagtgtg cgctgacggt
     4501 tccatcattg ccgtccagcc acggaacgtg tcctacgact ccgtgagcgc catcgtgacc
     4561 gcaaacctgt ccattccctt caactggacc acctcagtcc aagtggagta cctccagatt
     4621 accagcactc ccattgtcgt cgactgctcc acctacgtgt gcaacggcaa cgtccgctgc
     4681 gtggagctcc tgaagcagta cacttccgcc tgcaagacta tagaggatgc actccgcaac
     4741 tcggccatgc tcgaatcggc cgatgtcagc gagatgctga ccttcgacaa aaaggccttt
     4801 actctcgcca atgtgtccag ctttggcgat tacaatctgt cctcggtgat cccctcactt
     4861 ccgagaagcg gatccagggt ggccggaaga agcgccattg aggacatctt gttctccaag
     4921 ctcgtgacca gcggcttggg aactgtggac gccgattaca agaagtgcac caagggtctg
     4981 tccattgccg acctggcttg cgcccaatac tacaacggca tcatggtgct gccaggagtc
     5041 gcggacgcag aacggatggc tatgtatact gggtcgctga tcggaggcat tgccctcggc
     5101 ggactgactt ccgccgcctc cattccgttc tcccttgcta ttcaatcacg cctgaactac
     5161 gtggcgctgc agacggacgt gctgcaggaa aaccaacgca tccttgcggc ctccttcaac
     5221 aaggccatga ccaatatcgt ggatgctttc actggagtga acgatgcgat cacgcagacc
     5281 agccaggctc tgcagactgt ggccaccgcg ctgaacaaga tccaggacgt cgtgaaccag
     5341 caggggaaca gtctgaacca cctgacttcc caactgcgcc agaatttcca ggccatcagc
     5401 agctcgattc aggctatcta cgaccggctc gatccacctc aggccgacca acaagtggac
     5461 cggctgatca ccggtagact cgccgctctg aatgtgttcg tgagccacac cctgaccaag
     5521 tacacggaag tccgcgcgag cagacagctg gcccagcaga aggtcaacga atgcgtgaag
     5581 tcccagtcca agcgctacgg gttctgcgga aacggaactc acatcttttc cctggtcaac
     5641 gctgcacctg aaggattggt gttcctgcac actgtcctgc tgcccaccca gtacaaagac
     5701 gtggaggctt ggtctgggct gtgtgtggat ggcatcaacg gctatgtgct gagacagcct
     5761 aatctcgccc tctacaagga agggaactac taccgcatca ctagccggat catgttcgag
     5821 ccccgaattc ccactattgc cgacttcgtg caaatcgaaa actgtaatgt cacattcgtg
     5881 aacatctcgc ggtccgaact gcaaaccatt gtcccggagt acatcgacgt gaacaagacg
     5941 ctgcaggaac tctcctacaa actgcctaac tataccgtgc ccgacctcgt ggtggaacag
     6001 tacaaccaga ctatcctgaa cttgacttcc gagatcagca ctctcgagaa caagtcggcc
     6061 gagctgaatt acaccgtgca aaagcttcag accctcatcg acaacattaa ctccacactt
     6121 gtggacctga aatggctcaa ccgggtcgaa accggttccg gttacattcc ggaagcccct
     6181 agggacggac aggcctacgt ccggaaggac ggagaatggg tgctgttatc cactttcctg
     6241 gggcggtcgt tggaggtgct gttccaaggt ccaggccacc accaccatca tcaccaccac
     6301 tccgcttggt cacaccctca attcgagaag ggcggcggat caggaggggg aggatccgga
     6361 ggctcagcct ggagccaccc acagttcgag aagtaatcaa ctttcttgta caaagttgtg
     6421 aaattctgca gatatccatc acactggcgg ccgctcgagc atgcatctag agggccctat
     6481 tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg
     6541 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     6601 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     6661 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     6721 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     6781 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     6841 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     6901 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     6961 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     7021 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     7081 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     7141 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     7201 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     7261 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     7321 aggtgtggaa agtccctagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     7381 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     7441 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     7501 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     7561 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     7621 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     7681 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     7741 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     7801 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     7861 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     7921 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     7981 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     8041 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     8101 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     8161 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     8221 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     8281 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     8341 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     8401 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     8461 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     8521 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     8581 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     8641 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     8701 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     8761 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     8821 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     8881 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     8941 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     9001 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     9061 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     9121 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     9181 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     9241 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     9301 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     9361 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     9421 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     9481 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     9541 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     9601 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     9661 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     9721 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     9781 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     9841 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     9901 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     9961 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
    10021 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
    10081 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
    10141 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
    10201 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
    10261 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
    10321 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
    10381 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
    10441 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
    10501 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
    10561 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
    10621 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
    10681 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
    10741 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
    10801 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
    10861 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
    10921 ctgacgtc
//
