LOCUS       sequence-362124-       13498 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  COX2-KO in MDCK.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13498)
  AUTHORS   Watabe T, Yamahira S, Takakura K, Thumkeo D, Narumiya S, Matsuda M, 
            Terai K
  TITLE     Calcium transients trigger switch-like discharge of prostaglandin 
            E(2) in an extracellular signal-regulated kinase-dependent manner.
  JOURNAL   eLife. 2024 Jan 26;12:RP86727. doi: 10.7554/eLife.86727.
  PUBMED    38276879
REFERENCE   2  (bases 1 to 13498)
  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: "eLife."; 
            date: "2024-01-26"; volume: "12:RP86727. doi"; pages: " 
            10.7554/eLife.86727"
COMMENT     Sequence Label: LentiCRISPRv2-hyg2-COX2-4
FEATURES             Location/Qualifiers
     source          1..13498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        177..556
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        558..756
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     711..731
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             774..954
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1001..1126
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1619..1852
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2037..2081
                     /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    2379..2496
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2547..2795
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2547..2567
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2718..2737
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2817..2892
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2954..3165
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3184..3193
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3190..7293
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             3190..7293
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7294..7341
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7342..7365
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    7405..7972
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7566..7583)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7793..7812
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             8011..9030
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     LTR             9189..9422
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9448..9465)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9454..9678
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9724..10152
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9811..9830)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10021..10042
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10161..10181)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10166..10495
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10346..10481
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10408..10427
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10543..10590
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10609..10983
                     /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"
     polyA_signal    11113..11234
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11150..11169)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11204..11223
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11283..11299)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11283..11299)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11296..11318)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11307..11323
                     /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(11331..11337,11338..11355,11356..11361))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11376..11397
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11514..11531)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11685..12273)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11765..11784)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12444..13235,13236..13304))
                     /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     13067..13086
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13305..13409)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(join(13484..13498,1..5))
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
       61 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      121 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      181 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      241 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      301 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      361 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      421 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      481 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      541 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      601 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      661 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      721 cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc
      781 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      841 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      901 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      961 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
     1021 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
     1081 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
     1141 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
     1201 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct
     1261 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
     1321 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
     1381 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
     1441 aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga
     1501 gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag
     1561 tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag
     1621 gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga
     1681 cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc
     1741 tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc
     1801 tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt
     1861 ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta
     1921 ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta
     1981 acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga
     2041 atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa
     2101 caaattggct gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa
     2161 gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat
     2221 cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag
     2281 aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatcg gcactgcgtg
     2341 cgccaattct gcagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt
     2401 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
     2461 gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga
     2521 gatccagttt ggttaattaa ggtaccgagg gcctatttcc catgattcct tcatatttgc
     2581 atatacgata caaggctgtt agagagataa ttagaattaa tttgactgta aacacaaaga
     2641 tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa
     2701 aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct
     2761 tggctttata tatcttgtgg aaaggacgaa acaccgaagt tcttcgaatg atgtatgttt
     2821 tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca
     2881 ccgagtcggt gcttttttga attcgctagc taggtcttga aaggagtggg aattggctcc
     2941 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
     3001 gtcggcaatt gatccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
     3061 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
     3121 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggaccg gttctagagc
     3181 gctgccacca tggacaagaa gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc
     3241 tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac
     3301 accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga cagcggcgaa
     3361 acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac
     3421 cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc
     3481 ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc
     3541 atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac
     3601 ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc
     3661 ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac
     3721 aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag
     3781 gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc
     3841 aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg
     3901 ttcggaaacc tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac
     3961 ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac
     4021 ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc
     4081 gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg
     4141 agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct
     4201 ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac
     4261 ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa gttcatcaag
     4321 cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac
     4381 ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga
     4441 gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg
     4501 gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg
     4561 ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac
     4621 ttcgaggaag tggtggacaa gggcgcttcc gcccagagct tcatcgagcg gatgaccaac
     4681 ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac
     4741 ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc
     4801 gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg
     4861 aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc
     4921 gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg
     4981 ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa
     5041 gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa
     5101 acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc
     5161 ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag
     5221 acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc
     5281 cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc
     5341 gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc
     5401 ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag
     5461 aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc
     5521 cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa
     5581 gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag
     5641 aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat
     5701 gtggaccata tcgtgcctca gagctttctg aaggacgact ccatcgacaa caaggtgctg
     5761 accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag
     5821 aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca gagaaagttc
     5881 gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc
     5941 aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc
     6001 cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa agtgatcacc
     6061 ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag
     6121 atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg
     6181 atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac
     6241 gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc
     6301 ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc
     6361 cggaagcggc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg ggataagggc
     6421 cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag
     6481 accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgat
     6541 aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc
     6601 accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg
     6661 aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat
     6721 cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag
     6781 ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc
     6841 ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac
     6901 ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg
     6961 tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag cgagttctcc
     7021 aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta caacaagcac
     7081 cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat
     7141 ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc
     7201 agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag
     7261 acacggatcg acctgtctca gctgggaggc gacaagcgac ctgccgccac aaagaaggct
     7321 ggacaggcta agaagaagaa agattacaaa gacgatgacg ataagggctc tagaggatcc
     7381 cggtttccct ctagcgggat caattccgcc ccccccccct aacgttactg gccgaagccg
     7441 cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt
     7501 tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct
     7561 ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct
     7621 ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc
     7681 acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc
     7741 ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc
     7801 ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc
     7861 tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct
     7921 aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat aagcttgcca
     7981 caacccgtac caaagatgga tagatccgga aagcctgaac tcaccgcgac gtctgtcgag
     8041 aagtttctga tcgaaaagtt cgacagcgtg tccgacctga tgcagctctc ggagggcgaa
     8101 gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc
     8161 tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc
     8221 ccgattccgg aagtgcttga cattggggag ttcagcgaga gcctgaccta ttgcatctcc
     8281 cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg
     8341 cagccggtcg cggaggccat ggatgcgatc gctgcggccg atcttagcca gacgagcggg
     8401 ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc
     8461 gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg
     8521 tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg
     8581 cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca
     8641 gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc
     8701 ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt cgagcggagg
     8761 catccggagc ttgcaggatc gccgcggctc cgggcgtata tgctccgcat tggtcttgac
     8821 caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga
     8881 tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga
     8941 agcgcggccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc
     9001 cccagcactc gtccgagggc aaaggaatag agggcctgct gccggctctg cggcctcttc
     9061 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcgtc
     9121 gactttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     9181 ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac
     9241 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     9301 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     9361 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     9421 cagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     9481 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     9541 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     9601 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     9661 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     9721 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     9781 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     9841 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     9901 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     9961 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
    10021 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
    10081 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
    10141 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
    10201 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
    10261 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
    10321 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
    10381 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
    10441 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
    10501 cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg
    10561 gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagttg
    10621 accagtgccg ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc
    10681 gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac
    10741 gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc
    10801 tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg
    10861 aacttccggg acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg
    10921 gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac
    10981 tgacacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
    11041 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
    11101 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
    11161 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
    11221 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
    11281 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
    11341 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
    11401 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
    11461 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
    11521 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
    11581 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
    11641 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
    11701 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
    11761 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
    11821 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
    11881 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
    11941 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
    12001 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
    12061 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
    12121 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
    12181 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
    12241 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
    12301 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
    12361 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
    12421 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
    12481 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
    12541 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
    12601 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
    12661 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
    12721 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
    12781 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
    12841 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
    12901 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
    12961 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
    13021 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
    13081 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
    13141 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
    13201 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
    13261 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
    13321 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
    13381 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
    13441 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctga
//
