LOCUS       sequence-366917-        9805 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible nuclear cGAS with D307A mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9805)
  AUTHORS   Wu Y, Song K, Hao W, Li J, Wang L, Li S
  TITLE     Nuclear soluble cGAS senses double-stranded DNA virus infection.
  JOURNAL   Commun Biol. 2022 May 10;5(1):433. doi: 10.1038/s42003-022-03400-1.
  PUBMED    35538147
REFERENCE   2  (bases 1 to 9805)
  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: "Commun 
            Biol."; date: "2022-05-10"; volume: "5(1):433. doi"; pages: " 
            10.1038/s42003-022-03400-1"
COMMENT     Sequence Label: pCW57-mcGAS-LL/RK-NLS/D307A
FEATURES             Location/Qualifiers
     source          1..9805
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        37..351
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    45..63
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    81..99
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    116..134
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    152..170
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    188..206
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    223..241
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    259..277
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     324..348
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      368..377
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1898..1924
                     /codon_start=1
                     /product="nuclear localization signal (Makkerh et al., 
                     1996)"
                     /label=NLS
                     /translation="PAAKKKKLD"
     CDS             1946..2017
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     CDS             2039..2095
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     promoter        2123..2633
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2552..2570
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2651..3673
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             2651..3673
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3689..3742
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(3743..4360,4361..4366,4367..4402,4403..4405,4406..4441,
                     4442..4444,4445..4480,4481..4489)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(4529..4549)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4947..4958)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5163..5396
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5468..5589
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5505..5524)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5559..5578
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5629..5764
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5764..5835
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5836..5904,5905..6696)
                     /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     complement(6054..6073)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6867..7455
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7356..7375
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7609..7626
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7743..7764
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7779..7784,7785..7802,7803..7809)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7817..7833
                     /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)."
     primer_bind     7822..7844
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7841..7857
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7841..7857
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7876..7896
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7878..7896
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7924..8150
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             8151..8331
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8378..8503
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8996..9229
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9414..9458
                     /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    9671..9788
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 cagcagagat ccactttggc cgcgaatcga tatgtcgagt ttactcccta tcagtgatag
       61 agaacgtatg tcgagtttac tccctatcag tgatagagaa cgatgtcgag tttactccct
      121 atcagtgata gagaacgtat gtcgagttta ctccctatca gtgatagaga acgtatgtcg
      181 agtttactcc ctatcagtga tagagaacgt atgtcgagtt tatccctatc agtgatagag
      241 aacgtatgtc gagtttactc cctatcagtg atagagaacg tatgtcgagg taggcgtgta
      301 cggtgggagg cctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagaatt
      361 ggctagcgcc accatggaag atccgcgtag aaggacgacg gcgccacgcg ctaagaagcc
      421 gtccgcgaag cgcgccccga cgcagcccag caggaccagg gcccacgcgg aaagctgcgg
      481 cccgcaaagg ggggctcgat cgcggcgggc ggagcgtgac ggggacacca cggagaagcc
      541 acgtgcccca gggccccgag tgcatccagc aagggccact gagctcacca aagatgcaca
      601 gccctcggcc atggacgcgg caggagccac cgcgcggcct gccgtccggg tgccccagca
      661 gcaggccatc ctggatccgg agctgcccgc cgtacgggag ccccagccgc ccgcggatcc
      721 cgaggcgcgg aaagtcgtaa ggggacctag ccacagaagg ggcgcgcgct ccaccgggca
      781 gcccagagcg ccgcgagggt ccaggaagga accggacaag ctaaagaagg tgctggacaa
      841 aaggagaaag aaacgcaaag atatctcgga ggcggccgag acggtgaata aagttgtgga
      901 acgcctgctg cgcagaatgc agaaacggga gtcggagttc aaaggtgtgg agcagctgaa
      961 cactggcagc tactatgaac atgtgaagat ttctgctcct aatgaatttg atgttatgtt
     1021 taaactggaa gtccccagga ttgagctaca agaatattat gaaacaggtg ctttctatct
     1081 tgtgaaattc aaaagaattc cacgaggaaa tccgctgagt catttcttag aaggggaagt
     1141 attatcagct accaagatgc tgtcaaagtt taggaaaatc attaaagaag aagttaaaga
     1201 aatcaaagat atagatgtca gtgtggagaa ggaaaaacca ggaagccctg ctgtaacact
     1261 tcttatcagg aaccctgaag aaatctctgt ggctataatt ctggctttgg agtcaaaagg
     1321 cagctggcct attagtacca aagaaggact acctattcaa ggctggctgg gcacaaaagt
     1381 gaggaccaat ctaagacgag agccgtttta tctcgtaccc aagaatgcaa aggatggaaa
     1441 tagttttcaa ggagagacct ggcgcctctc tttctctcac actgaaaagt acattttgaa
     1501 taatcacggg atagagaaaa catgctgtga atcttccgga gcaaaatgct gcagaaaaga
     1561 atgtttaaaa ttaatgaaat accttttgga acagttgaaa aaagagtttc aagagctgga
     1621 tgcattctgt tcctaccatg tgaaaactgc catctttcac atgtggaccc aggacccgca
     1681 ggacagtcag tgggacccca ggaacctcag ctcctgcttc gataagttgt tagcattctt
     1741 tcttgagtgc ctcaggacag agaaactgga tcattatttt attccaaagt tcaatctatt
     1801 ctctcaagaa ctaattgacc gaaaaagtaa agaatttcta tcgaagaaaa ttgaatatga
     1861 aagaaataat gggtttccaa tttttgacaa gcttagcccg gcagctaaga aaaagaaact
     1921 ggatgcggcc gcaatcgatg tcgaggatta caaggatgac gacgataagg actataagga
     1981 cgatgatgac aaggactaca aagatgatga cgataaatag gtcgacaccg gttccggagc
     2041 cacgaacttc tctctgttaa agcaagcagg agacgtggaa gaaaaccccg gtcctaggct
     2101 gcagacgcgt tgtacaggat ccggggttgg ggttgcgcct tttccaaggc agccctgggt
     2161 ttgcgcaggg acgcggctgc tctgggcgtg gttccgggaa acgcagcggc gccgaccctg
     2221 ggtctcgcac attcttcacg tccgttcgca gcgtcacccg gatcttcgcc gctacccttg
     2281 tgggcccccc ggcgacgctt cctgctccgc ccctaagtcg ggaaggttcc ttgcggttcg
     2341 cggcgtgccg gacgtgacaa acggaagccg cacgtctcac tagtaccctc gcagacggac
     2401 agcgccaggg agcaatggca gcgcgccgac cgcgatgggc tgtggccaat agcggctgct
     2461 cagcagggcg cgccgagagc agcggccggg aaggggcggt gcgggaggcg gggtgtgggg
     2521 cggtagtgtg ggccctgttc ctgcccgcgc ggtgttccgc attctgcaag cctccggagc
     2581 gcacgtcggc agtcggctcc ctcgttgacc gaatcaccga cctctctccc cagcaattca
     2641 cccgcgcgcg atgaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga
     2701 aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt
     2761 cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt
     2821 ctacaaagat cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt
     2881 gcttgacatt ggggaattta gcgagagcct gacctattgc atctcccgcc gtgcacaggg
     2941 tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga
     3001 ggccatggat gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg
     3061 accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc
     3121 ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc
     3181 tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc
     3241 ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg
     3301 gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc
     3361 gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc
     3421 aggatcgccg cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag
     3481 cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt
     3541 ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg
     3601 gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc
     3661 gagggcaaag gaattcgaag gaagcggaga aggtagaggt tctctcctca cttgtggtga
     3721 tgttgaagaa aaccctggtc caatgtctag actggacaag agcaaagtca taaacggagc
     3781 tctggaatta ctcaatggtg tcggtatcga aggcctgacg acaaggaaac tcgctcaaaa
     3841 gctgggagtt gagcagccta ccctgtactg gcacgtgaag aacaagcggg ccctgctcga
     3901 tgccctgcca atcgagatgc tggacaggca tcatacccac ttctgccccc tggaaggcga
     3961 gtcatggcaa gactttctgc ggaacaacgc caagtcatac cgctgtgctc tcctctcaca
     4021 tcgcgacggg gctaaagtgc atctcggcac ccgcccaaca gagaaacagt acgaaaccct
     4081 ggaaaatcag ctcgcgttcc tgtgtcagca aggcttctcc ctggagaacg cactgtacgc
     4141 tctgtccgcc gtgggccact ttacactggg ctgcgtattg gaggaacagg agcatcaagt
     4201 agcaaaagag gaaagagaga cacctaccac cgattctatg cccccacttc tgagacaagc
     4261 aattgagctg ttcgaccggc agggagccga acctgccttc cttttcggcc tggaactaat
     4321 catatgtggc ctggagaaac agctaaagtg cgaaagcggc gggccgaccg acgcccttga
     4381 cgattttgac ttagacatgc tcccagccga tgcccttgac gactttgacc ttgatatgct
     4441 gcctgctgac gctcttgacg attttgacct tgacatgctc cccgggtaac taagtaagga
     4501 tcgatccaag atatcgtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     4561 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     4621 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     4681 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     4741 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     4801 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     4861 tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc
     4921 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     4981 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     5041 tctccctttg ggccgcctcc ccgcctgttt cgcctcggcg tccggactag aggtaccttt
     5101 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg
     5161 actggaaggg ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc
     5221 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     5281 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     5341 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag
     5401 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag
     5461 tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5521 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5581 tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccat cccgccccta
     5641 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     5701 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     5761 ggcctttcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5821 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5881 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5941 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     6001 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     6061 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     6121 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     6181 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     6241 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     6301 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     6361 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     6421 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     6481 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     6541 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     6601 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     6661 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     6721 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6781 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6841 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6901 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6961 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     7021 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     7081 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     7141 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     7201 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     7261 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     7321 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     7381 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     7441 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     7501 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     7561 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     7621 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     7681 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     7741 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     7801 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     7861 gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctgga gctgcaagct
     7921 taatgtagtc ttatgcaata ctcttgtagt cttgcaacat ggtaacgatg agttagcaac
     7981 atgccttaca aggagagaaa aagcaccgtg catgccgatt ggtggaagta aggtggtacg
     8041 atcgtgcctt attaggaagg caacagacgg gtctgacatg gattggacga accactgaat
     8101 tgccgcattg cagagatatt gtatttaagt gcctagctcg atacataaac gggtctctct
     8161 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     8221 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     8281 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc
     8341 gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct
     8401 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
     8461 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
     8521 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
     8581 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
     8641 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
     8701 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
     8761 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
     8821 taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga gatatgaggg
     8881 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     8941 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     9001 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc
     9061 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     9121 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     9181 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     9241 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     9301 aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca
     9361 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg
     9421 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa
     9481 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa
     9541 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt
     9601 ttcagaccca cctcccaacc ccgaggggac aattctcgac ctcgagacaa atggcagtat
     9661 tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag
     9721 tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc
     9781 aaaattttcg ggtttattac aggga
//
