LOCUS       sequence-362122-       13355 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  COX1-KO in MDCK.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13355)
  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 13355)
  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-bleo-COX1-4
FEATURES             Location/Qualifiers
     source          1..13355
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(38..57)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        229..608
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        610..808
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     763..783
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             826..1006
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1053..1178
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1671..1904
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2089..2133
                     /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    2431..2548
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2599..2847
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2599..2619
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2770..2789
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2869..2944
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3006..3217
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3236..3245
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3242..7345
                     /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             3242..7345
                     /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             7346..7393
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7394..7417
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    7496..8045
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7640..7657)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7867..7886
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             8048..8422
                     /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"
     misc_feature    8438..9026
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8491..8511)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8909..8920)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9098..9331
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9357..9374)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9363..9587
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9633..10061
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9720..9739)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9930..9951
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10070..10090)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10075..10404
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10255..10390
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10317..10336
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10452..10499
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10518..10892
                     /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    11022..11143
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11059..11078)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11113..11132
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11192..11208)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11192..11208)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11205..11227)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11216..11232
                     /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(11240..11246,11247..11264,11265..11270))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11285..11306
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11423..11440)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11594..12182)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11674..11693)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12353..13144,13145..13213))
                     /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     12976..12995
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13214..13318)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat
       61 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
      121 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg
      181 ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat
      241 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
      301 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      361 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      421 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      481 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      541 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      601 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      661 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      721 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      781 gtgtacggtg ggaggtctat ataagcagcg cgttttgcct gtactgggtc tctctggtta
      841 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
      901 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
      961 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
     1021 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
     1081 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     1141 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     1201 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     1261 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
     1321 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     1381 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     1441 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     1501 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat
     1561 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     1621 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     1681 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     1741 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     1801 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     1861 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     1921 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     1981 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
     2041 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
     2101 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
     2161 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
     2221 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
     2281 acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga
     2341 gagagagaca gagacagatc cattcgatta gtgaacggat cggcactgcg tgcgccaatt
     2401 ctgcagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta
     2461 cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca
     2521 aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt
     2581 ttggttaatt aaggtaccga gggcctattt cccatgattc cttcatattt gcatatacga
     2641 tacaaggctg ttagagagat aattagaatt aatttgactg taaacacaaa gatattagta
     2701 caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt
     2761 tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta
     2821 tatatcttgt ggaaaggacg aaacaccgcc gtgacatgct catgcgtcgt tttagagcta
     2881 gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
     2941 gtgctttttt gaattcgcta gctaggtctt gaaaggagtg ggaattggct ccggtgcccg
     3001 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
     3061 ttgatccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
     3121 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
     3181 cgttcttttt cgcaacgggt ttgccgccag aacacaggac cggttctaga gcgctgccac
     3241 catggacaag aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt
     3301 gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg
     3361 gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga
     3421 ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg
     3481 ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag
     3541 actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg
     3601 caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa
     3661 gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca
     3721 catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga
     3781 cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc
     3841 catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag
     3901 acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa
     3961 cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga
     4021 ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc
     4081 ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat
     4141 cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc
     4201 tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg
     4261 gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc
     4321 cggctacatt gacggcggag ccagccagga agagttctac aagttcatca agcccatcct
     4381 ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg
     4441 gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca
     4501 cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat
     4561 cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag
     4621 cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga
     4681 agtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa
     4741 gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt
     4801 gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct
     4861 gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac
     4921 cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat
     4981 ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat
     5041 tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt
     5101 gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc
     5161 ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg
     5221 caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct
     5281 ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga
     5341 cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct
     5401 gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac
     5461 agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt
     5521 gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag
     5581 aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc
     5641 cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg
     5701 ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg atgtggacca
     5761 tatcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag
     5821 cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa
     5881 gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct
     5941 gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca
     6001 gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa
     6061 cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc
     6121 caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa
     6181 ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa
     6241 gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa
     6301 gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag
     6361 caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg
     6421 gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt
     6481 tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt
     6541 gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat
     6601 cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc
     6661 ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt
     6721 gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga
     6781 ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa
     6841 gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact
     6901 gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag
     6961 ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga
     7021 acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt
     7081 gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa
     7141 gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc
     7201 ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa
     7261 agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat
     7321 cgacctgtct cagctgggag gcgacaagcg acctgccgcc acaaagaagg ctggacaggc
     7381 taagaagaag aaagattaca aagacgatga cgataagggc tctagaggat ccgttaacta
     7441 acttaagcta gcaacggttt ccctctagcg ggatcaattc cgcccccccc ccctaacgtt
     7501 actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc
     7561 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc
     7621 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag
     7681 gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg
     7741 cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat
     7801 acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga
     7861 gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc
     7921 cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg
     7981 ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga
     8041 taataccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg
     8101 agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt
     8161 cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt
     8221 gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg
     8281 gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg
     8341 cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt
     8401 cgtggccgag gagcaggact gaacgcgtta agtcgacaat caacctctgg attacaaaat
     8461 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     8521 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     8581 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     8641 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     8701 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     8761 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     8821 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     8881 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     8941 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     9001 gatctccctt tgggccgcct ccccgcgtcg actttaagac caatgactta caaggcagct
     9061 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     9121 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     9181 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     9241 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     9301 cccttttagt cagtgtggaa aatctctagc agggcccgtt taaacccgct gatcagcctc
     9361 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     9421 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     9481 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     9541 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     9601 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     9661 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     9721 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     9781 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     9841 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     9901 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     9961 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
    10021 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
    10081 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
    10141 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
    10201 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
    10261 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
    10321 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
    10381 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
    10441 gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca
    10501 aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg
    10561 acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg
    10621 aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg
    10681 accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt
    10741 acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga
    10801 ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact
    10861 gcgtgcactt cgtggccgag gagcaggact gacacgtgct acgagatttc gattccaccg
    10921 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
    10981 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
    11041 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
    11101 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
    11161 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
    11221 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
    11281 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
    11341 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
    11401 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
    11461 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
    11521 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
    11581 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
    11641 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
    11701 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
    11761 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
    11821 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
    11881 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
    11941 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
    12001 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
    12061 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
    12121 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
    12181 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
    12241 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
    12301 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
    12361 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
    12421 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
    12481 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
    12541 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
    12601 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
    12661 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
    12721 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
    12781 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
    12841 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
    12901 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
    12961 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
    13021 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
    13081 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
    13141 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
    13201 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
    13261 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
    13321 catttccccg aaaagtgcca cctgacgtcg acgga
//
