LOCUS       sequence-256941-       13406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycyclin inducible shRNA knockdown of mouse cGAS gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13406)
  AUTHORS   Vanpouille-Box C, Alard A, Aryankalayil MJ, Sarfraz Y, Diamond JM, 
            Schneider RJ, Inghirami G, Coleman CN, Formenti SC, Demaria S
  TITLE     DNA exonuclease Trex1 regulates radiotherapy-induced tumour 
            immunogenicity.
  JOURNAL   Nat Commun. 2017 Jun 9;8:15618. doi: 10.1038/ncomms15618.
  PUBMED    28598415
REFERENCE   2  (bases 1 to 13406)
  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: "2017-06-9"; volume: "8:15618. doi"; pages: " 
            10.1038/ncomms15618"
COMMENT     Sequence Label: pTRIPZ shcGAS-Mmu
FEATURES             Location/Qualifiers
     source          1..13406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2063..2180
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(2245..2619)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     regulatory      2616..2625
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2629..2647)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2639..2686)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     protein_bind    2722..2740
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2757..2775
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2793..2811
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2829..2847
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2864..2882
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2900..2918
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        2930..2968
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2965..2989
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3005..3700
                     /codon_start=1
                     /product="red fluorescent protein from Entacmaea 
                     quadricolor"
                     /label=TurboRFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
     ncRNA           3713..3834
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           3862..3876
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           3904..4030
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     promoter        complement(4125..4143)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        4156..5367
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5204..5223
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    complement(5380..5413)
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are 
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     CDS             5420..6127
                     /codon_start=1
                     /product="reverse tetracycline transactivator 3"
                     /label=rtTA3
                     /note="activates expression from Tet-responsive promoters 
                     in the presence of doxycycline"
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPG"
     protein_bind    complement(6128..6161)
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are 
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     misc_feature    6184..6756
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6351..6368)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6578..6597
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6759..7361
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MATEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGSRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     7258..7278
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7377..7965
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7430..7450)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7848..7859)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8172..8405
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8506..8523)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8512..8736
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8782..9210
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8869..8888)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9079..9100
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9219..9239)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9224..9553
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9404..9539
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9466..9485
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    10761..10882
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10931..10947)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10931..10947)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10944..10966)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10955..10971
                     /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(10979..10985,10986..11003,11004..11009))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11024..11045
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11162..11179)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11333..11921)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11413..11432)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12092..12883,12884..12952))
                     /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     12715..12734
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12953..13057)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    13105..13239
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac
     2041 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
     2101 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
     2161 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttag
     2221 taccgggccc gctctagtcc ggaatcagtc ctgctcctcg gccacgaagt gcacgcagtt
     2281 gccggccggg tcgcgcaggg cgaactcccg cccccacggc tgctcgccga tctcggtcat
     2341 ggccggcccg gaggcgtccc ggaagttcgt ggacacgacc tccgaccact cggcgtacag
     2401 ctcgtccagg ccgcgcaccc acacccaggc cagggtgttg tccggcacca cctggtcctg
     2461 gaccgcgctg atgaacaggg tcacgtcgtc ccggaccaca ccggcgaagt cgtcctccac
     2521 gaagtcccgg gagaacccga gccggtcggt ccagaactcg accgctccgg cgacgtcgcg
     2581 cgcggtgagc accggaacgg cactggtcaa cttggccatg gtggccctcc tatagtgagt
     2641 cgtattatac tatgccgata tactatgccg atgattaatt gtcaacacgt gctgcaggtc
     2701 cgaggttcta gacgagttta ctccctatca gtgatagaga acgatgtcga gtttactccc
     2761 tatcagtgat agagaacgta tgtcgagttt actccctatc agtgatagag aacgtatgtc
     2821 gagtttactc cctatcagtg atagagaacg tatgtcgagt ttatccctat cagtgataga
     2881 gaacgtatgt cgagtttact ccctatcagt gatagagaac gtatgtcgag gtaggcgtgt
     2941 acggtgggag gcctatataa gcagagctcg tttagtgaac cgtcagatcg caccggtcgc
     3001 caccatgagc gagctgatca aggagaacat gcacatgaag ctgtacatgg agggcaccgt
     3061 gaacaaccac cacttcaagt gcacatccga gggcgaaggc aagccctacg agggcaccca
     3121 gaccatgaag atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg acatcctggc
     3181 taccagcttc atgtacggca gcaaagcctt catcaaccac acccagggca tccccgactt
     3241 ctttaagcag tccttccctg agggcttcac atgggagaga atcaccacat acgaagacgg
     3301 gggcgtgctg accgctaccc aggacaccag cttccagaac ggctgcatca tctacaacgt
     3361 caagatcaac ggggtgaact tcccatccaa cggccctgtg atgcagaaga aaacacgcgg
     3421 ctgggaggcc aacaccgaga tgctgtaccc cgctgacggc ggcctgagag gccacagcca
     3481 gatggccctg aagctcgtgg gcgggggcta cctgcactgc tccttcaaga ccacatacag
     3541 atccaagaaa cccgctaaga acctcaagat gcccggcttc cacttcgtgg accacagact
     3601 ggaaagaatc aaggaggccg acaaagagac ctacgtcgag cagcacgaga tggctgtggc
     3661 caagtactgc gacctcccta gcaaactggg gcacagataa tcgatagttt gtttgaatga
     3721 ggcttcagta ctttacagaa tcgttgcctg cacatcttgg aaacacttgc tgggattact
     3781 tcttcaggtt aacccaacag aaggctcgag aaggtatatt gctgttgaca gtgagcgaag
     3841 gattgagcta caagaatatt agtgaagcca cagatgtaat attcttgtag ctcaatcctg
     3901 tgcctactgc ctcggaattc aaggggctac tttaggagca attatcttgt ttactaaaac
     3961 tgaatacctt gctatctctt tgatacattt ttacaaagct gaattaaaat ggtataaatt
     4021 aaatcacttt tttcaattgg aagactaatg cggccggcca ttactccgtc tcgtgtcttg
     4081 ttgcatatgt ctgctggttt gtttgatgtt gtttgcgggc gggccctata gtgagtcgta
     4141 ttacctagga cgcgtggcct ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcc
     4201 tcacggcgag cgctgccacg tcagacgaag ggcgcagcga gcgtcctgat ccttccgccc
     4261 ggacgctcag gacagcggcc cgctgctcat aagactcggc cttagaaccc cagtatcagc
     4321 agaaggacat tttaggacgg gacttgggtg actctagggc actggttttc tttccagaga
     4381 gcggaacagg cgaggaaaag tagtcccttc tcggcgattc tgcggaggga tctccgtggg
     4441 gcggtgaacg ccgatgatta tataaggacg cgccgggtgt ggcacagcta gttccgtcgc
     4501 agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta
     4561 gcgggctgct gggctggccg gggctttcgt ggccgccggg ccgctcggtg ggacggaagc
     4621 gtgtggagag accgccaagg gctgtagtct gggtccgcga gcaaggttgc cctgaactgg
     4681 gggttggggg gagcgcagca aaatggcggc tgttcccgag tcttgaatgg aagacgcttg
     4741 tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg caagaaccca
     4801 aggtcttgag gccttcgcta atgcgggaaa gctcttattc gggtgagatg ggctggggca
     4861 ccatctgggg accctgacgt gaagtttgtc actgactgga gaactcggtt tgtcgtctgt
     4921 tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca cccgtacctt tgggagcgcg
     4981 cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg gtggggccac
     5041 ctgccggtag gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc gggcctaggg
     5101 taggctctcc tgaatcgaca ggcgccggac ctctggtgag gggagggata agtgaggcgt
     5161 cagtttcttt ggtcggtttt atgtacctat cttcttaagt agctgaagct ccggttttga
     5221 actatgcgct cggggttggc gagtgtgttt tgtgaagttt tttaggcacc ttttgaaatg
     5281 taatcatttg ggtcaatatg taattttcag tgttagacta gtaaattgtc cgctaaattc
     5341 tggccgtttt tggctttttt gttagacgct agcggatcca taacttcgta tagtatacat
     5401 tatacgaagt tatgccacca tgtctaggct ggacaagagc aaagtcataa acggagctct
     5461 ggaattactc aatggtgtcg gtatcgaagg cctgacgaca aggaaactcg ctcaaaagct
     5521 gggagttgag cagcctaccc tgtactggca cgtgaagaac aagcgggccc tgctcgatgc
     5581 cctgccaatc gagatgctgg acaggcatca tacccacttc tgccccctgg aaggcgagtc
     5641 atggcaagac tttctgcgga acaacgccaa gtcataccgc tgtgctctcc tctcacatcg
     5701 cgacggggct aaagtgcatc tcggcacccg cccaacagag aaacagtacg aaaccctgga
     5761 aaatcagctc gcgttcctgt gtcagcaagg cttctccctg gagaacgcac tgtacgctct
     5821 gtccgccgtg ggccacttta cactgggctg cgtattggag gaacaggagc atcaagtagc
     5881 aaaagaggaa agagagacac ctaccaccga ttctatgccc ccacttctga gacaagcaat
     5941 tgagctgttc gaccggcagg gagccgaacc tgccttcctt ttcggcctgg aactaatcat
     6001 atgtggcctg gagaaacagc taaagtgcga aagcggcggg ccgaccgacg cccttgacga
     6061 ttttgactta gacatgctcc cagccgatgc ccttgacgat tttgaccttg acatgctccc
     6121 cgggtaaata acttcgtata gtatacatta tacgaagtta tggatccgcg gccgcaaatt
     6181 ccgcccctct ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg
     6241 gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc
     6301 ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa
     6361 aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag
     6421 acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg
     6481 cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg
     6541 ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa
     6601 caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg
     6661 gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca
     6721 cggggacgtg gttttccttt gaaaaacacg ataataccat ggccaccgag tacaagccca
     6781 cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt
     6841 tcgccgacta ccccgccacg cgccacaccg tcgacccgga ccgccacatc gagcgggtca
     6901 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     6961 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     7021 tgttcgccga gatcggctcg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     7081 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     7141 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     7201 gagtggaggc ggccgagcgc gctggggtgc ccgccttcct ggagacctcc gcgccccgca
     7261 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     7321 gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg agttcgcgtc tggaacaatc
     7381 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     7441 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     7501 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     7561 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     7621 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     7681 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     7741 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct
     7801 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     7861 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     7921 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga attaattctg
     7981 cagtcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc
     8041 tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca
     8101 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     8161 aaagagggga ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg
     8221 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     8281 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     8341 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     8401 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     8461 atcagagagt gagaggcctt gacattgttt aaacccgctg atcagcctcg actgtgcctt
     8521 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     8581 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     8641 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     8701 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     8761 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     8821 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     8881 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     8941 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     9001 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     9061 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     9121 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     9181 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     9241 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     9301 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     9361 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     9421 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     9481 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     9541 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt
     9601 gatgaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga
     9661 cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga
     9721 tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga
     9781 tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg attccggaag tgcttgacat
     9841 tggggaatta attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca
     9901 cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca
     9961 tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc
    10021 aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg
    10081 tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg
    10141 atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt
    10201 tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg
    10261 aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt
    10321 tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat
    10381 cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg
    10441 ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat
    10501 ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg
    10561 atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg
    10621 caaaggaata gcacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg
    10681 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
    10741 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
    10801 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
    10861 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
    10921 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
    10981 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
    11041 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
    11101 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
    11161 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
    11221 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
    11281 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
    11341 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
    11401 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
    11461 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
    11521 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
    11581 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
    11641 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
    11701 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
    11761 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
    11821 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
    11881 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
    11941 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
    12001 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
    12061 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
    12121 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
    12181 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
    12241 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
    12301 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
    12361 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
    12421 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
    12481 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
    12541 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
    12601 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
    12661 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
    12721 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
    12781 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
    12841 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
    12901 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
    12961 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
    13021 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
    13081 ctgacgtcga cggatcggga gatcaacttg tttattgcag cttataatgg ttacaaataa
    13141 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
    13201 ttgtccaaac tcatcaatgt atcttatcat gtctggatca actggataac tcaagctaac
    13261 caaaatcatc ccaaacttcc caccccatac cctattacca ctgccaatta cctgtggttt
    13321 catttactct aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt
    13381 taaatatgta ctacaaactt agtagt
//
