LOCUS       sequence-422110-       12319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of SARS-CoV-2 NSP16 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12319)
  AUTHORS   Broderick K, Moutaoufik MT, Saccon T, Malty R, Amin S, Phanse S, 
            Joseph TP, Zilocchi M, Hosseinnia A, Istace Z, Hajikarimlou M, Abrar
            S, Fisher J, Brassard R, Perera R, Kumar A, Aoki H, Rahmatbakhsh M, 
            Jessulat M, Kobasa D, Dehne F, Prasad B, Gagarinova A, Joanne 
            Lemieux M, Cochrane A, Houry WA, Aly KA, Golshani A, Babu M
  TITLE     Human protein interaction networks of ancestral and variant 
            SARS-CoV-2 in organ-specific cells and bodily fluids.
  JOURNAL   Nat Commun. 2025 Jul 1;16(1):5784. doi: 10.1038/s41467-025-60949-1.
  PUBMED    40593736
REFERENCE   2  (bases 1 to 12319)
  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: "Nat 
            Commun."; date: "2025-07-1"; volume: "16(1):5784. doi"; pages: " 
            10.1038/s41467-025-60949-1"
COMMENT     Sequence Label: pLD-puro-Cc-SARS-CoV-2_NSP16-VA
FEATURES             Location/Qualifiers
     source          1..12319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(162..750)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(921..1712,1713..1781))
                     /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     1544..1563
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1782..1886)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1912..2367)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2049..2070)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2261..2280
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2509..2525
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2535..2553
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      2574..2709
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2749..2870
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2760..2779)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2814..2833
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             2942..3175
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     CDS             complement(3303..3902)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3386..3406)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3883..3902
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(3924..4434)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3987..4005)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     misc_feature    4483..4600
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(4648..4713)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(4726..4746)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(4759..4779)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(4789..4806)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4819..4842)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(4879..4902)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    4982..5006
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5904..6003)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(6019..6035)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      6099..6185
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6278..6305
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      complement(6635..7223)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7316..8107)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     primer_bind     8538..8554
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8559..8577
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    8595..8694
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(9592..9616)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(9647..9671)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(9668..9871)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(9697..9717)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(10160..10180)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     CDS             complement(10478..10522)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    10707..10940
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    11433..11558
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             11605..11785
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(11786..12012)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(12040..12060)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(12040..12058)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(12079..12095)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12092..12114)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12103..12119
                     /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(12127..12133,12134..12151,12152..12157))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12172..12193
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
       61 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
      121 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
      181 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      241 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      301 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      361 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      421 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      481 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
      541 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      601 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
      661 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
      721 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
      781 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
      841 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
      901 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
      961 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     1021 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     1081 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     1141 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     1201 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     1261 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     1321 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     1381 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     1441 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     1501 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     1561 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     1621 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     1681 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     1741 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     1801 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     1861 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
     1921 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     1981 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     2041 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     2101 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     2161 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     2221 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     2281 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     2341 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg
     2401 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     2461 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     2521 ccagtgagcg cgcgtaatac gactcactat agggcgaatt gggtacgtcc ctaggcctcc
     2581 aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc
     2641 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
     2701 gggcgggatg ggcggagtta ggggcgggat agctagagcc agacatgata agatacattg
     2761 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     2821 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt cctctcactc
     2881 tctgatattc atttctttgc aagttataaa tactgaataa taagatgaca tgaactacta
     2941 ctgctagaga ttttccacac tgactaaaag ggtctgaggg atctctagtt accagagtca
     3001 cacaacagac gggcacacac tacttgaagc actcaaggca agctttattg aggcttaagc
     3061 agtgggttcc ctagttagcc agagagctcc caggctcaga tctggtctaa ccagagagac
     3121 ccagtacaag caaaaagcag atcttgtctt cgttgggagt gaattagccc ttccagtccc
     3181 cccttttctt ttaaaaagtg gctaagatct acagctgcct tgtaagtcat tggtcttaaa
     3241 ggtaccgatg catggggtcg tgcgctcctt tcggtcgggc gctgcgggtc gtggggcggg
     3301 cgtcaggcac cgggcttgcg ggtcatgcac caggtgcgcg gtccttcggg cacctcgacg
     3361 tcggcggtga cggtgaagcc gagccgctcg tagaagggga ggttgcgggg cgcggaggtc
     3421 tccaggaagg cgggcacccc ggcgcgctcg gccgcctcca ctccggggag cacgacggcg
     3481 ctgcccagac ccttgccctg gtggtcgggc gagacgccga cggtggccag gaaccacgcg
     3541 ggctccttgg gccggtgcgg cgccaggagg ccttccatct gttgctgcgc ggccagccgg
     3601 gaaccgctca actcggccat gcgcgggccg atctcggcga acaccgcccc cgcttcgacg
     3661 ctctccggcg tggtccagac cgccaccgcg gcgccgtcgt ccgcgaccca caccttgccg
     3721 atgtcgagcc cgacgcgcgt gaggaagagt tcttgcagct cggtgacccg ctcgatgtgg
     3781 cggtccggat cgacggtgtg gcgcgtggcg gggtagtcgg cgaacgcggc ggcgagggtg
     3841 cgtacggccc tggggacgtc gtcgcgggtg gcgaggcgca ccgtgggctt gtactcggtc
     3901 atggtaagct ccggtggatc cccctgggga gagaggtcgg tgattcggtc aacgagggag
     3961 ccgactgccg acgtgcgctc cggaggcttg cagaatgcgg aacaccgcgc gggcaggaac
     4021 agggcccaca ctaccgcccc acaccccgcc tcccgcaccg ccccttcccg gccgctgctc
     4081 tcggcgcgcc ctgctgagca gccgctattg gccacagccc atcgcggtcg gcgcgctgcc
     4141 attgctccct ggcgctgtcc gtctgcgagg gtactagtga gacgtgcggc ttccgtttgt
     4201 cacgtccggc acgccgcgaa ccgcaaggaa ccttcccgac ttaggggcgg agcaggaagc
     4261 gtcgccgggg ggcccacaag ggtagcggcg aagatccggg tgacgctgcg aacggacgtg
     4321 aagaatgtgc gagacccagg gtcggcgccg ctgcgtttcc cggaaccacg cccagagcag
     4381 ccgcgtccct gcgcaaaccc agggctgcct tggaaaaggc gcaaccccaa ccccctcgag
     4441 ccgcggccaa agtggatctc tgctgtccct gtaataaacc cgaaaatttt gaatttttgt
     4501 aatttgtttt tgtaattctt tagtttgtat gtctgttgct attatgtcta ctattctttc
     4561 ccctgcactg taccccccaa tccccccttt tcttttaaaa ttgtggatga atactgccat
     4621 ttgtctcgag gtcgagaatt cgaactactt gtcatcgtca tccttgtaat cgatgtcatg
     4681 atctttataa tcaccgtcat ggtctttgta gtctcttcct gcaggcccct gaaagtacag
     4741 gttctcgctg cctcctctcc cctggaagta cagattttct cctgcagggt gatggtgatg
     4801 gtgatggccg cttccgcctt tctcgaactg cgggtggctc cagcttcctc ctccgctgcc
     4861 tccgccgctt cctcctccct tctcgaactg agggtggctc caggcgatcg ccttgatcac
     4921 cacggggggc tggttctcgc ccagcaggtt gaacagctct ctggcgatcg ctctagaatc
     4981 aaccactttg tacaagaaag ttgggtagtt gttcaccagc acgtcagagg agatgaccac
     5041 tctgttgttt tccctgatga tcaggcgtcc cttggacagc aggctcagga tcatgtcgtt
     5101 gatctgtccc tccttcaggc tcatcacagc ggtgcctctc agcttcagtg ggaacttaga
     5161 catgtcgaac agggagtagc tagacagctg gatagggttg gtgttgcgcc agaagatgta
     5221 gttggcgtgc atcacgtaac cgtcgatctg ttcacggggc tttcccaggt agttgcagcc
     5281 gatcaggaaa gcctcggagc tagaggcgtt cacgttggtc acgaaagcgg tccaccaggc
     5341 gaagtgtccc atcagcttgt acaggtcagc gttccaggag tgttcggtga tcttgatagc
     5401 cacagatccg cccagggcca gcttctgctg gatgaagccg cagatgtagg tgaagaaacc
     5461 ttccttgctg tcgttctcct tggtcacgtt cttggtcttt gggtcgtaca tgtcagagat
     5521 gatcaggtcc cacttgttag cggtgtgcac agtagcacag tcgccgatca gggtgctgtc
     5581 agcatcagac acgaagtcgt tcaggtcaga gtccaccagc agggtaccgg tgggcagcca
     5641 ctgtctcagc acagcggtac ctggagccac tcccttgtca gaaccagcac cgaagtggat
     5701 cacgcgcatg ttgtaaggca cggccagggt cagggtgttc aggtactggc acagctgggt
     5761 gtacttagcc acgttcatca tgataccctt gggcagggtg gcggagtctc cgtagttctg
     5821 caggtcgcac ttctccagca gcatacgctg catcttgtac aggttaggca tagccactcc
     5881 aggctgccaa gcctgggaag acatgccaac ttttttgtac aaagttggca ttataaaaaa
     5941 gcattgctca tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     6001 ttggggcccg agcttaagac tggccgtcgt tttacaacgt cgtgactggg aaaacatcca
     6061 tgctagcgtt aacgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt
     6121 cggaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     6181 caaatccgcc gggagcggat ttgaacgttg tgaagcaacg gcccggaggg ntggcgggca
     6241 ggacgcccgc cataaactgc caggcatcaa actaagcaga aggccatcct gacggatggc
     6301 ctttttgcgt ttctacaaac tcttcctggc tagcggtacg cgtattaatt gcgttgcgct
     6361 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     6421 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     6481 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     6541 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     6601 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     6661 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6721 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6781 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6841 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6901 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6961 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     7021 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     7081 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     7141 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     7201 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     7261 agtggaacga cgcgtaactc acgttaaggg attttggtca tgggtggctc gagggttatt
     7321 tgccgactac cttggtgatc tcgcctttca cgtagtggac aaattcttcc aactgatctg
     7381 cgcgcgaggc caagcgatct tcttcttgtc caagataagc ctgtctagct tcaagtatga
     7441 cgggctgata ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg
     7501 cgattttgcc ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct
     7561 catcgccagc ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa
     7621 atagatcctg ttcaggaacc ggatcaaaga gttcctccgc cgctggacct accaaggcaa
     7681 cgctatgttc tcttgctttt gtcagcaaga tagccagatc aatgtcgatc gtggctggct
     7741 cgaagatacc tgcaagaatg tcattgcgct gccattctcc aaattgcagt tcgcgcttag
     7801 ctggataacg ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga
     7861 gaatctcgct ctctccaggg gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc
     7921 gcgttgtttc atcaagcctt acggtcaccg taaccagcaa atcaatatca ctgtgtggct
     7981 tcaggccgcc atccactgcg gagccgtaca aatgtacggc cagcaacgtc ggttcgagat
     8041 ggcgctcgat gacgccaact acctctgata gttgagtcga tacttcggcg atcaccgctt
     8101 ccctcataat gtttaacttt gttttagggc gactgccctg ctgcgtaaca tcgttgctgc
     8161 tccataacat caaacatcga cccacggcgt aacgcgcttg ctgcttggat gcccgaggca
     8221 tagactgtac cccaaaaaaa cagtcataac aagccatgaa aaccgccact gcgccgttac
     8281 caccgctgcg ttcggtcaag gttctggacc agttgcgtga gcgcatacgc tacttgcatt
     8341 acagcttacg aaccgaacag gcttatgtcc actgggttcg tgccttcatc cgtttccacg
     8401 gtgtgcgtca cccggcaacc ttgggtagca gcgaagtcga ggcatttctg tcctggctgg
     8461 tctagaggag gcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag
     8521 acacgggcca gagctgccag gaaacagcta tgaccatgta atacgactca ctatagggga
     8581 tatcagctgg atggcaaata atgattttat tttgactgat agtgacctgt tcgttgcaac
     8641 aaattgataa gcaatgcttt cttataatgc caactttgta caagaaagtt gggtagttgt
     8701 tcaccagcac gtcagaggag atgaccactc tgttgttttc cctgatgatc aggcgtccct
     8761 tggacagcag gctcaggatc atgtcgttga tctgtccctc cttcaggctc atcacagcgg
     8821 tgcctctcag cttcagtggg aacttagaca tgtcgaacag ggagtagcta gacagctgga
     8881 tagggttggt gttgcgccag aagatgtagt tggcgtgcat cacgtaaccg tcgatctgtt
     8941 cacggggctt tcccaggtag ttgcagccga tcaggaaagc ctcggagcta gaggcgttca
     9001 cgttggtcac gaaagcggtc caccaggcga agtgtcccat cagcttgtac aggtcagcgt
     9061 tccaggagtg ttcggtgatc ttgatagcca cagatccgcc cagggccagc ttctgctgga
     9121 tgaagccgca gatgtaggtg aagaaacctt ccttgctgtc gttctccttg gtcacgttct
     9181 tggtctttgg gtcgtacatg tcagagatga tcaggtccca cttgttagcg gtgtgcacag
     9241 tagcacagtc gccgatcagg gtgctgtcag catcagacac gaagtcgttc aggtcagagt
     9301 ccaccagcag ggtaccggtg ggcagccact gtctcagcac agcggtacct ggagccactc
     9361 ccttgtcaga accagcaccg aagtggatca cgcgcatgtt gtaaggcacg gccagggtca
     9421 gggtgttcag gtactggcac agctgggtgt acttagccac gttcatcatg atacccttgg
     9481 gcagggtggc ggagtctccg tagttctgca ggtcgcactt ctccagcagc atacgctgca
     9541 tcttgtacag gttaggcata gccactccag gctgccaagc ctgggaagac atgccaactt
     9601 ttttgtacaa acttgtacgc gtacagcgtg gatggcgtct ccaggcgatc tgacggttca
     9661 ctaaacgagc tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca
     9721 atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac
     9781 tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac
     9841 gcccattgat gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga
     9901 tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc
     9961 atttaccgtc attgacgtca atagggggcg tacttggcat atgatagtcc attttaaaac
    10021 ataattttaa aactgcaaac tacccaagaa attattactt tctacgtcac gtattttgta
    10081 ctaatatctt tgtgtttaca gtcaaattaa ttccaattat ctctctaaca gccttgtatc
    10141 gtatatgcaa atatgaagga atcatgggaa ataggccctc ggtgaagggg gcggccgctc
    10201 gaggctagtc tcgtgatcga taccgtcgag atccgttcac taatcgaatg gatctgtctc
    10261 tgtctctctc tccaccttct tcttctattc cttcgggcct gtcgggtccc ctcggggttg
    10321 ggaggtgggt ctgaaacgat aatggtgaat atccctgcct aactctattc actatagaaa
    10381 gtacagcaaa aactattctt aaacctacca agcctcctac tatcattatg aataatttta
    10441 tataccacag ccaatttgtt atgttaaacc aattccacaa acttgcccat ttatctaatt
    10501 ccaataattc ttgttcattc ttttcttgct ggttttgcga ttcttcaatt aaggagtgta
    10561 ttaagcttgt gtaattgtta atttctctgt cccactccat ccaggtcgtg tgattccaaa
    10621 tctgttccag agatttatta ctccaactag cattccaagg cacagcagtg gtgcaaatga
    10681 gttttccaga gcaaccccaa atccccagga gctgttgatc ctttaggtat ctttccacag
    10741 ccaggattct tgcctggagc tgcttgatgc cccagactgt gagttgcaac agatgctgtt
    10801 gcgcctcaat agccctcagc aaattgttct gctgctgcac tataccagac aataattgtc
    10861 tggcctgtac cgtcagcgtc attgacgctg cgcccatagt gcttcctgct gctcccaaga
    10921 acccaaggaa caaagctcct attcccactg ctcttttttc tctctgcacc actcttctct
    10981 ttgccttggt gggtgctact cctaatggtt caatttttac tactttatat ttatataatt
    11041 cacttctcca attgtccctc atatctcctc ctccaggtct gaagatcagc ggccgcttgc
    11101 tgtgcggtgg tcttactttt gttttgctct tcctctatct tgtctaaagc ttccttggtg
    11161 tcttttatct ctatcctttg atgcacacaa tagagggttg ctactgtatt atataatgat
    11221 ctaagttctt ctgatcctgt ctgaagggat ggttgtagct gtcccagtat ttgtctacag
    11281 ccttctgatg tttctaacag gccaggatta actgcgaatc gttctagctc cctgcttgcc
    11341 catactatat gttttaattt atattttttc tttccccctg gccttaaccg aattttttcc
    11401 catcgcgatc taattctccc ccgcttaata ctgacgctct cgcacccatc tctctccttc
    11461 tagcctccgc tagtcaaaat ttttggcgta ctcaccagtc gccgcccctc gcctcttgcc
    11521 gtgcgcgctt cagcaagccg agtcctgcgt cgagagagct cctctggttt ccctttcgct
    11581 ttcaagtccc tgttcgggcg ccactgctag agattttcca cactgactaa aagggtctga
    11641 gggatctcta gttaccagag tcacacaaca gacgggcaca cactacttga agcactcaag
    11701 gcaagcttta ttgaggctta agcagtgggt tccctagtta gccagagagc tcccaggctc
    11761 agatctggtc taaccagaga gacccgttta tgtatcgagc taggcactta aatacaatat
    11821 ctctgcaatg cggcaattca gtggttcgtc caatccatgt cagacccgtc tgttgccttc
    11881 ctaataaggc acgatcgtac caccttactt ccaccaatcg gcatgcacgg tgctttttct
    11941 ctccttgtaa ggcatgttgc taactcatcg ttaccatgtt gcaagactac aagagtattg
    12001 cataagacta cattaagctt gcagctccag cttttgttcc ctttagtgag ggttaattgc
    12061 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
    12121 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
    12181 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
    12241 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
    12301 ttccgcttcc tcgctcact
//
