LOCUS       sequence-259307-       13407 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycyclin inducible shRNA knockdown of human cGAS gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13407)
  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 13407)
  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-Hsa
FEATURES             Location/Qualifiers
     source          1..13407
                     /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..6757
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6352..6369)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6579..6598
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6760..7362
                     /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     7259..7279
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7378..7966
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7431..7451)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7849..7860)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8173..8406
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8507..8524)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8513..8737
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8783..9211
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8870..8889)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9080..9101
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9220..9240)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9225..9554
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9405..9540
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9467..9486
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    10762..10883
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10932..10948)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10932..10948)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10945..10967)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10956..10972
                     /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(10980..10986,10987..11004,11005..11010))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11025..11046
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11163..11180)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11334..11922)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11414..11433)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12093..12884,12885..12953))
                     /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     12716..12735
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12954..13058)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    13106..13240
                     /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 gtgagcgcag
     3841 gaaggaaatg gtttccaagt agtgaagcca cagatgtact tggaaaccat ttccttcctt
     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 ccccntaacg ttactggccg aagccgcttg gaataaggcc
     6241 ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg
     6301 cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca
     6361 aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa
     6421 gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt
     6481 gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt
     6541 gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca
     6601 acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc
     6661 ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc
     6721 acggggacgt ggttttcctt tgaaaaacac gataatacca tggccaccga gtacaagccc
     6781 acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg
     6841 ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat cgagcgggtc
     6901 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     6961 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     7021 gtgttcgccg agatcggctc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     7081 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     7141 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     7201 ggagtggagg cggccgagcg cgctggggtg cccgccttcc tggagacctc cgcgccccgc
     7261 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     7321 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gagttcgcgt ctggaacaat
     7381 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     7441 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     7501 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     7561 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     7621 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     7681 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     7741 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc
     7801 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     7861 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     7921 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg aattaattct
     7981 gcagtcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca gctaccaatg
     8041 ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca gtcacacctc
     8101 aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
     8161 aaaagagggg actggaaggg ctaattcact cccaacgaag acaagatctg ctttttgctt
     8221 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     8281 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     8341 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     8401 ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa
     8461 tatcagagag tgagaggcct tgacattgtt taaacccgct gatcagcctc gactgtgcct
     8521 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     8581 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     8641 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     8701 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     8761 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     8821 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     8881 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     8941 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     9001 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     9061 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     9121 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     9181 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     9241 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     9301 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     9361 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     9421 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     9481 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     9541 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg
     9601 tgatgaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg
     9661 acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg
     9721 atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag
     9781 atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca
     9841 ttggggaatt aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc
     9901 acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc
     9961 atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg
    10021 caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat
    10081 gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc
    10141 gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat
    10201 ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc
    10261 gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg
    10321 ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga
    10381 tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg
    10441 gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga
    10501 tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc
    10561 gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg
    10621 gcaaaggaat agcacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg
    10681 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
    10741 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
    10801 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
    10861 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
    10921 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
    10981 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
    11041 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
    11101 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
    11161 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
    11221 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
    11281 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
    11341 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
    11401 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
    11461 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
    11521 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
    11581 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
    11641 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
    11701 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
    11761 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
    11821 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
    11881 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
    11941 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
    12001 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
    12061 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
    12121 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
    12181 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
    12241 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
    12301 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
    12361 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
    12421 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
    12481 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
    12541 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
    12601 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
    12661 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
    12721 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
    12781 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
    12841 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
    12901 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
    12961 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
    13021 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
    13081 cctgacgtcg acggatcggg agatcaactt gtttattgca gcttataatg gttacaaata
    13141 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
    13201 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa
    13261 ccaaaatcat cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt
    13321 tcatttactc taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg
    13381 ttaaatatgt actacaaact tagtagt
//
