LOCUS       sequence-413637-       13013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  knock out WAVE2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13013)
  AUTHORS   Liu X, Nie L, Zhang Y, Yan Y, Wang C, Colic M, Olszewski K, Horbath 
            A, Chen X, Lei G, Mao C, Wu S, Zhuang L, Poyurovsky MV, James You M,
            Hart T, Billadeau DD, Chen J, Gan B
  TITLE     Actin cytoskeleton vulnerability to disulfide stress mediates 
            disulfidptosis.
  JOURNAL   Nat Cell Biol. 2023 Mar;25(3):404-414. doi: 
            10.1038/s41556-023-01091-2. Epub 2023 Feb 6.
  PUBMED    36747082
REFERENCE   2  (bases 1 to 13013)
  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"; doi: 
            "10.1038/s41556-023-01091-2"; journalName: "Nat Cell Biol"; date: 
            "2023-03"; volume: "25"; issue: "3"; pages: "404-414"
COMMENT     Sequence Label: lentiCRISPR v2-sgWAVE2-2
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     141..161
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             204..384
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    431..556
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1049..1282
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1467..1511
                     /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    1809..1926
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1977..2225
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1977..1997
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2148..2167
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2247..2322
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2384..2595
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2614..2623
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2620..6723
                     /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             2620..6723
                     /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             6724..6771
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6772..6795
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6805..6861
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             6862..7458
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     7355..7375
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7474..8062
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7527..7547)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7945..7956)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8134..8367
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8393..8410)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8399..8623
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8669..9097
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8756..8775)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8966..8987
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9106..9126)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9111..9440
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9291..9426
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9353..9372
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9488..9535
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9554..9928
                     /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    10058..10179
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10095..10114)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10149..10168
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10228..10244)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10228..10244)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10241..10263)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10252..10268
                     /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(10276..10282,10283..10300,10301..10306))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10321..10342
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10459..10476)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10630..11218)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10710..10729)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11389..12180,12181..12249))
                     /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     12012..12031
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12250..12354)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12429..12448)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        12620..12999
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(13001..13013,1..186)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
       61 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      121 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      181 aagcagcgcg ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc
      241 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
      301 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
      361 agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa
      421 accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga
      481 ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga
      541 tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg
      601 gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga
      661 gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat
      721 actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa
      781 tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc
      841 tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc
      901 tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat
      961 ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg
     1021 tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag
     1081 caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt
     1141 ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt
     1201 tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat
     1261 acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca
     1321 ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca
     1381 cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa
     1441 ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat
     1501 gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata aaattattca
     1561 taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga
     1621 atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg
     1681 gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca
     1741 ttcgattagt gaacggatcg gcactgcgtg cgccaattct gcagacaaat ggcagtattc
     1801 atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
     1861 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
     1921 aattttcggg tttattacag ggacagcaga gatccagttt ggttaattaa ggtaccgagg
     1981 gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt agagagataa
     2041 ttagaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt
     2101 aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc
     2161 ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa
     2221 acaccggggg taagattgtt gagagggttt tagagctaga aatagcaagt taaaataagg
     2281 ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcttttttga attcgctagc
     2341 taggtcttga aaggagtggg aattggctcc ggtgcccgtc agtgggcaga gcgcacatcg
     2401 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gatccggtgc ctagagaagg
     2461 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
     2521 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
     2581 gccgccagaa cacaggaccg gttctagagc gctgccacca tggacaagaa gtacagcatc
     2641 ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg
     2701 cccagcaaga aattcaaggt gctgggcaac accgaccggc acagcatcaa gaagaacctg
     2761 atcggagccc tgctgttcga cagcggcgaa acagccgagg ccacccggct gaagagaacc
     2821 gccagaagaa gatacaccag acggaagaac cggatctgct atctgcaaga gatcttcagc
     2881 aacgagatgg ccaaggtgga cgacagcttc ttccacagac tggaagagtc cttcctggtg
     2941 gaagaggata agaagcacga gcggcacccc atcttcggca acatcgtgga cgaggtggcc
     3001 taccacgaga agtaccccac catctaccac ctgagaaaga aactggtgga cagcaccgac
     3061 aaggccgacc tgcggctgat ctatctggcc ctggcccaca tgatcaagtt ccggggccac
     3121 ttcctgatcg agggcgacct gaaccccgac aacagcgacg tggacaagct gttcatccag
     3181 ctggtgcaga cctacaacca gctgttcgag gaaaacccca tcaacgccag cggcgtggac
     3241 gccaaggcca tcctgtctgc cagactgagc aagagcagac ggctggaaaa tctgatcgcc
     3301 cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc tgattgccct gagcctgggc
     3361 ctgaccccca acttcaagag caacttcgac ctggccgagg atgccaaact gcagctgagc
     3421 aaggacacct acgacgacga cctggacaac ctgctggccc agatcggcga ccagtacgcc
     3481 gacctgtttc tggccgccaa gaacctgtcc gacgccatcc tgctgagcga catcctgaga
     3541 gtgaacaccg agatcaccaa ggcccccctg agcgcctcta tgatcaagag atacgacgag
     3601 caccaccagg acctgaccct gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac
     3661 aaagagattt tcttcgacca gagcaagaac ggctacgccg gctacattga cggcggagcc
     3721 agccaggaag agttctacaa gttcatcaag cccatcctgg aaaagatgga cggcaccgag
     3781 gaactgctcg tgaagctgaa cagagaggac ctgctgcgga agcagcggac cttcgacaac
     3841 ggcagcatcc cccaccagat ccacctggga gagctgcacg ccattctgcg gcggcaggaa
     3901 gatttttacc cattcctgaa ggacaaccgg gaaaagatcg agaagatcct gaccttccgc
     3961 atcccctact acgtgggccc tctggccagg ggaaacagca gattcgcctg gatgaccaga
     4021 aagagcgagg aaaccatcac cccctggaac ttcgaggaag tggtggacaa gggcgcttcc
     4081 gcccagagct tcatcgagcg gatgaccaac ttcgataaga acctgcccaa cgagaaggtg
     4141 ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg
     4201 aaatacgtga ccgagggaat gagaaagccc gccttcctga gcggcgagca gaaaaaggcc
     4261 atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac
     4321 tacttcaaga aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga agatcggttc
     4381 aacgcctccc tgggcacata ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg
     4441 gacaatgagg aaaacgagga cattctggaa gatatcgtgc tgaccctgac actgtttgag
     4501 gacagagaga tgatcgagga acggctgaaa acctatgccc acctgttcga cgacaaagtg
     4561 atgaagcagc tgaagcggcg gagatacacc ggctggggca ggctgagccg gaagctgatc
     4621 aacggcatcc gggacaagca gtccggcaag acaatcctgg atttcctgaa gtccgacggc
     4681 ttcgccaaca gaaacttcat gcagctgatc cacgacgaca gcctgacctt taaagaggac
     4741 atccagaaag cccaggtgtc cggccagggc gatagcctgc acgagcacat tgccaatctg
     4801 gccggcagcc ccgccattaa gaagggcatc ctgcagacag tgaaggtggt ggacgagctc
     4861 gtgaaagtga tgggccggca caagcccgag aacatcgtga tcgaaatggc cagagagaac
     4921 cagaccaccc agaagggaca gaagaacagc cgcgagagaa tgaagcggat cgaagagggc
     4981 atcaaagagc tgggcagcca gatcctgaaa gaacaccccg tggaaaacac ccagctgcag
     5041 aacgagaagc tgtacctgta ctacctgcag aatgggcggg atatgtacgt ggaccaggaa
     5101 ctggacatca accggctgtc cgactacgat gtggaccata tcgtgcctca gagctttctg
     5161 aaggacgact ccatcgacaa caaggtgctg accagaagcg acaagaaccg gggcaagagc
     5221 gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga actactggcg gcagctgctg
     5281 aacgccaagc tgattaccca gagaaagttc gacaatctga ccaaggccga gagaggcggc
     5341 ctgagcgaac tggataaggc cggcttcatc aagagacagc tggtggaaac ccggcagatc
     5401 acaaagcacg tggcacagat cctggactcc cggatgaaca ctaagtacga cgagaatgac
     5461 aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc cgatttccgg
     5521 aaggatttcc agttttacaa agtgcgcgag atcaacaact accaccacgc ccacgacgcc
     5581 tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt accctaagct ggaaagcgag
     5641 ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag
     5701 gaaatcggca aggctaccgc caagtacttc ttctacagca acatcatgaa ctttttcaag
     5761 accgagatta ccctggccaa cggcgagatc cggaagcggc ctctgatcga gacaaacggc
     5821 gaaaccgggg agatcgtgtg ggataagggc cgggattttg ccaccgtgcg gaaagtgctg
     5881 agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa
     5941 gagtctatcc tgcccaagag gaacagcgat aagctgatcg ccagaaagaa ggactgggac
     6001 cctaagaagt acggcggctt cgacagcccc accgtggcct attctgtgct ggtggtggcc
     6061 aaagtggaaa agggcaagtc caagaaactg aagagtgtga aagagctgct ggggatcacc
     6121 atcatggaaa gaagcagctt cgagaagaat cccatcgact ttctggaagc caagggctac
     6181 aaagaagtga aaaaggacct gatcatcaag ctgcctaagt actccctgtt cgagctggaa
     6241 aacggccgga agagaatgct ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc
     6301 ctgccctcca aatatgtgaa cttcctgtac ctggccagcc actatgagaa gctgaagggc
     6361 tcccccgagg ataatgagca gaaacagctg tttgtggaac agcacaagca ctacctggac
     6421 gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga cgctaatctg
     6481 gacaaagtgc tgtccgccta caacaagcac cgggataagc ccatcagaga gcaggccgag
     6541 aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt caagtacttt
     6601 gacaccacca tcgaccggaa gaggtacacc agcaccaaag aggtgctgga cgccaccctg
     6661 atccaccaga gcatcaccgg cctgtacgag acacggatcg acctgtctca gctgggaggc
     6721 gacaagcgac ctgccgccac aaagaaggct ggacaggcta agaagaagaa agattacaaa
     6781 gacgatgacg ataagggatc cggcgcaaca aacttctctc tgctgaaaca agccggagat
     6841 gtcgaagaga atcctggacc gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     6901 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     6961 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     7021 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     7081 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     7141 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg
     7201 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggagtctc gcccgaccac
     7261 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc
     7321 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     7381 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     7441 cgcaagcccg gtgcctgaac gcgttaagtc gacaatcaac ctctggatta caaaatttgt
     7501 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     7561 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     7621 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     7681 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     7741 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     7801 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     7861 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     7921 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     7981 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     8041 tccctttggg ccgcctcccc gcgtcgactt taagaccaat gacttacaag gcagctgtag
     8101 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     8161 gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     8221 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     8281 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     8341 tttagtcagt gtggaaaatc tctagcaggg cccgtttaaa cccgctgatc agcctcgact
     8401 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     8461 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     8521 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     8581 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     8641 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     8701 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     8761 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     8821 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     8881 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     8941 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     9001 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     9061 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     9121 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     9181 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     9241 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     9301 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     9361 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     9421 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     9481 agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     9541 gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt
     9601 cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact tcgtggagga
     9661 cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg tccaggacca
     9721 ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc
     9781 cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga
     9841 gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt
     9901 gcacttcgtg gccgaggagc aggactgaca cgtgctacga gatttcgatt ccaccgccgc
     9961 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
    10021 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
    10081 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
    10141 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
    10201 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
    10261 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
    10321 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
    10381 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
    10441 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
    10501 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
    10561 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
    10621 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
    10681 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
    10741 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
    10801 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
    10861 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
    10921 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
    10981 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
    11041 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
    11101 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
    11161 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
    11221 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
    11281 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
    11341 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
    11401 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
    11461 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
    11521 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
    11581 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
    11641 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
    11701 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
    11761 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
    11821 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
    11881 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
    11941 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
    12001 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
    12061 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
    12121 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
    12181 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
    12241 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
    12301 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
    12361 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
    12421 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
    12481 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
    12541 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
    12601 cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg
    12661 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
    12721 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
    12781 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
    12841 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
    12901 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
    12961 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttg
//
