LOCUS       sequence-327662-       11432 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of HCoV-HKU1 soluble spike trimer protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11432)
  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 11432)
  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-M89-303: CMV51p> HCoV-HKU1
            S-2P-T4f-3C-Hi...
FEATURES             Location/Qualifiers
     source          1..11432
                     /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             6754..6777
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             6781..6804
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             6811..6834
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             6874..6897
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    complement(6899..6923)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(6985..7003)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6986..7003)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(7023..7040)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7029..7253
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7299..7727
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7386..7405)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7596..7617
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7736..7756)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7741..8070
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7921..8056
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7983..8002
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             8137..8931
                     /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(8191..8210)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8801..8820
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    9105..9226
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9142..9161)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9196..9215
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9275..9291)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9275..9291)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9288..9310)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9299..9315
                     /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(9323..9329,9330..9347,9348..9353))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9368..9389
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9506..9523)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9677..10265)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9757..9776)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10436..11227,11228..11296))
                     /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     11059..11078
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11297..11401)
                     /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 cattttcatc ctaccgacta ccctggccgt gatcggagac
     2881 ttcaactgca ctaactcctt tatcaacgac tacaacaaga ctattccgcg catttccgag
     2941 gacgtggtgg acgtgtcact gggcctgggc acttactacg tgctgaaccg ggtgtacctg
     3001 aataccaccc tcctgtttac cggatacttc cctaagtcgg gggcgaactt ccgggatctg
     3061 gctctgaagg gatccatcta cctctctacc ctgtggtaca agccgccgtt cctgtccgac
     3121 ttcaataacg gcatcttcag caaggtcaag aacaccaagc tctacgtgaa caacaccctc
     3181 tactccgagt tctccacaat cgtcattggt tcagtgttcg tgaacactag ctacacaatc
     3241 gtggtgcagc cccataacgg tattctggag atcaccgcct gccaatacac tatgtgcgag
     3301 taccctcaca ccgtgtgtaa atcaaagggc agcatcagaa acgagagctg gcacatcgat
     3361 tccagcgaac ccctctgcct cttcaaaaag aacttcacct acaacgtgtc agccgactgg
     3421 ctctacttcc acttctacca agagcgcgga gtgttctacg cctattacgc cgacgtgggc
     3481 atgcctacta ccttcctctt ctccctgtac ctgggtacta tcctctccca ttactacgtg
     3541 atgccgctga cctgtaacgc gatcagcagc aacactgata atgagacttt ggaatattgg
     3601 gtcacgcctt tgtcgaggcg gcagtatctg ctgaacttcg acgaacacgg cgtgatcacc
     3661 aacgccgtgg actgcagcag ctccttcctg tcggagattc agtgcaagac ccagtccttt
     3721 gcgcctaaca ccggagtgta cgatctctcc ggcttcactg tgaaacctgt ggccaccgtc
     3781 taccgccgca ttcccaacct ccccgattgc gacatcgaca actggctgaa caacgtgtcc
     3841 gtgccatcgc cgctgaactg ggagcggagg atcttctcga actgcaactt caatctgtca
     3901 actctgctgc ggctggtgca cgtggactcc ttttcctgca acaacctgga caagtcgaag
     3961 attttcggat cctgctttaa cagcatcacc gtcgacaagt tcgccatccc gaaccggaga
     4021 agggatgatc ttcagcttgg gtcctccggg ttcttgcagt cctccaacta caagatcgat
     4081 atcagttcct catcctgcca actgtactac tcgttgccgc tcgtgaatgt gaccattaac
     4141 aacttcaacc caagctcctg gaaccggcgc tacggattcg gtagctttaa cctgtcgtcc
     4201 tacgatgtgg tgtactcaga ccactgtttc tcggtcaact ccgacttctg cccatgcgct
     4261 gacccttccg tcgtgaattc ctgcgccaag tcgaagccgc cttcggcgat ttgtcctgct
     4321 ggaaccaagt atcggcactg cgacctggat actactctgt acgtaaagaa ctggtgccgc
     4381 tgctcctgtc tgcccgaccc catctccact tactccccga acacgtgccc ccagaagaag
     4441 gtggtcgtgg gaatcggaga gcactgtcca ggattgggaa ttaacgagga aaaatgcggc
     4501 acccagctga accacagcag ctgcttctgc tcccctgacg cattcctggg atggtccttc
     4561 gattcatgca tttccaacaa ccggtgtaac atattctcca acttcatctt caacgggatc
     4621 aattccggca ccacttgctc gaacgatctg ctatattcca acaccgagat ctccaccggc
     4681 gtctgcgtga actacgacct ctacggcatc acgggtcaag gcatcttcaa ggaagtcagc
     4741 gcggcctact acaacaactg gcagaacttg ctgtacgaca gcaacggaaa catcattggt
     4801 ttcaaggact tcctgactaa caagacctac accatccttc cctgctactc cgggagagtc
     4861 tccgccgcct tctaccaaaa ctcctctagc cccgcccttc tgtatcgcaa cctgaagtgc
     4921 agctacgtcc tcaacaacat atcgttcatc agccagcctt tctactttga ctcctacctg
     4981 ggctgcgtgc tcaacgcagt gaatcttacc tcatattcgg tcagcagctg tgacctccgc
     5041 atgggctcag ggttctgcat cgattacgca ctcccgtcct ccggtggaag cggcagcggc
     5101 atctcatccc cctaccggtt cgtgaccttc gaacccttca acgtgtcctt cgtgaacgac
     5161 tccgtggaaa ccgtcggagg gctgttcgag atccagatcc ctaccaattt caccattgcc
     5221 ggacacgagg aattcatcca aaccagctcg cccaaggtca ccatcgactg ttccgccttt
     5281 gtgtgctcca attacgcggc atgccacgat cttctgtccg aatacggaac cttctgcgat
     5341 aacatcaact ccattctcaa cgaggtcaat gatctgctgg atattaccca gttacaagtg
     5401 gccaacgccc tgatgcaggg cgtgactctg tcctccaacc tgaatactaa cctccattcg
     5461 gatgtggaca acatcgactt caagtcgttg ctgggctgcc tgggaagcca atgcggatcc
     5521 tcctcacggt cgctgctgga agatctgctg tttaacaagg ttaagctgag cgacgtggga
     5581 tttgtggaag cctataacaa ctgtaccggg ggatccgaga tccgggacct cctctgcgtg
     5641 cagtcgttca acggtatcaa ggtcctgccc cctattctga gcgagactca gatttcggga
     5701 tacactaccg ctgcgactgt ggcagctatg ttccctccgt ggtctgccgc tgctggagtg
     5761 ccattctccc tgaacgtcca gtaccgcatc aatggactgg gagtgaccat ggatgtgctg
     5821 aacaagaacc agaagctcat cgccaacgcc ttcaacaagg ccctgctgag catccagaac
     5881 ggattcacgg ccaccaattc cgccctggcc aaaatccaat cagtcgtgaa cgctaacgcc
     5941 caggccctga acagcctgct ccagcagctg ttcaacaagt tcggcgccat ttcctcctcg
     6001 ctccaagaaa ttctgtcacg gctggatccg cctgaagcac aggtccagat tgacagactc
     6061 atcaatgggc gactgactgc cctgaacgcc tacgtcagcc aacagttgag cgacatcacc
     6121 ttgattaagg ccggagcctc cagagctatc gaaaaagtca acgaatgcgt caagtcccag
     6181 tcgccacgga tcaacttctg cggaaatgga aaccatattc tgtccctcgt gcaaaacgcc
     6241 ccgtacggac tgttgttcat ccacttctca tacaagccaa cctcgttcaa gaccgtgctg
     6301 gtgtccccgg gactctgtct ctccggggat cggggaattg cgccgaagca ggggtacttc
     6361 attaagcaga acgactcctg gatgttcaca ggatcttcct actactaccc cgaacccatc
     6421 tccgacaaga acgtcgtgtt tatgaattcg tgcagcgtca actttaccaa ggcccccttc
     6481 atctacttga acaactcgat cccgaacctc agcgacttcg aagccgagct cagcttatgg
     6541 ttcaagaacc acacctccat cgcacccaat ctgaccttca actcgcatat caacgcgacc
     6601 ttcttggacc tttactatga gatgaacgtg atccaggaaa gcatcaagtc ccttaatggc
     6661 tccggctaca tccctgaagc accgagggac ggtcaagcct acgtgcgcaa ggacggggaa
     6721 tgggtgctcc tgtccacgtt cctgggtcgc tccctggagg tgctgttcca gggaccgggg
     6781 catcaccacc accatcatca ccactcagcc tggtcacacc cccaattcga aaagggagga
     6841 ggttcgggcg gtggaggaag cggtggctct gcgtggtccc acccacaatt cgagaagtaa
     6901 tcaactttct tgtacaaagt tgtgaaattc tgcagatatc catcacactg gcggccgctc
     6961 gagcatgcat ctagagggcc ctattctata gtgtcaccta aatgctagag ctcgctgatc
     7021 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     7081 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     7141 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     7201 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     7261 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     7321 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     7381 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     7441 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     7501 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     7561 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     7621 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     7681 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     7741 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     7801 atgcatctca attagtcagc aaccaggtgt ggaaagtccc taggctcccc agcaggcaga
     7861 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     7921 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     7981 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     8041 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     8101 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     8161 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     8221 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     8281 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     8341 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     8401 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     8461 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     8521 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     8581 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     8641 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     8701 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     8761 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     8821 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     8881 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     8941 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     9001 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     9061 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     9121 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     9181 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     9241 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     9301 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     9361 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     9421 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     9481 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     9541 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     9601 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     9661 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     9721 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     9781 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     9841 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     9901 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     9961 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
    10021 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
    10081 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
    10141 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
    10201 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
    10261 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
    10321 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
    10381 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
    10441 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
    10501 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
    10561 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
    10621 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
    10681 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
    10741 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
    10801 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
    10861 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
    10921 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
    10981 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
    11041 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
    11101 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
    11161 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
    11221 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
    11281 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
    11341 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
    11401 gcacatttcc ccgaaaagtg ccacctgacg tc
//
