LOCUS       sequence-208130-       13012 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses sgRNA targeting mouse Kif26b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13012)
  AUTHORS   Susman MW, Karuna EP, Kunz RC, Gujral TS, Cantu AV, Choi SS, Jong 
            BY, Okada K, Scales MK, Hum J, Hu LS, Kirschner MW, Nishinakamura R,
            Yamada S, Laird DJ, Jao LE, Gygi SP, Greenberg ME, Ho HH
  TITLE     Kinesin superfamily protein Kif26b links Wnt5a-Ror signaling to the 
            control of cell and tissue behaviors in vertebrates.
  JOURNAL   Elife. 2017 Sep 8;6. pii: e26509. doi: 10.7554/eLife.26509.
  PUBMED    28885975
REFERENCE   2  (bases 1 to 13012)
  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: "2017-09-8"; volume: "6. pii: e26509. doi"; pages: " 
            10.7554/eLife.26509"
COMMENT     Sequence Label: lentiCRISPR v2 kif26b sgRNA1
FEATURES             Location/Qualifiers
     source          1..13012
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(40..59)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        231..610
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        612..810
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     765..785
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             828..1008
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1055..1180
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1673..1906
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2091..2135
                     /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    2433..2550
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2601..2849
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2601..2621
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2772..2791
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2870..2945
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3007..3218
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3237..3246
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3243..7346
                     /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             3243..7346
                     /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             7347..7394
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7395..7418
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             7428..7484
                     /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             7485..8081
                     /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     7978..7998
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8097..8685
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8150..8170)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8568..8579)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8757..8990
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9016..9033)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9022..9246
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9292..9720
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9379..9398)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9589..9610
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9729..9749)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9734..10063
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9914..10049
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9976..9995
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10111..10158
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10177..10551
                     /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    10681..10802
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10718..10737)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10772..10791
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10851..10867)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10851..10867)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10864..10886)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10875..10891
                     /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(10899..10905,10906..10923,10924..10929))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10944..10965
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11082..11099)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11253..11841)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11333..11352)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12012..12803,12804..12872))
                     /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     12635..12654
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12873..12977)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc
       61 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
      121 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
      181 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt
      241 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
      301 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      361 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      421 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      481 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      541 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      601 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      661 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      721 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      781 gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg tctctctggt
      841 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
      901 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
      961 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
     1021 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
     1081 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
     1141 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
     1201 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
     1261 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
     1321 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
     1381 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
     1441 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
     1501 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
     1561 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     1621 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     1681 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     1741 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     1801 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     1861 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     1921 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     1981 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     2041 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     2101 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     2161 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     2221 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     2281 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
     2341 gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa
     2401 ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg
     2461 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta
     2521 caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca
     2581 gtttggttaa ttaaggtacc gagggcctat ttcccatgat tccttcatat ttgcatatac
     2641 gatacaaggc tgttagagag ataattagaa ttaatttgac tgtaaacaca aagatattag
     2701 tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat
     2761 gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt
     2821 tatatatctt gtggaaagga cgaaacaccg cttacgagga gtcgcgcgcg ttttagagct
     2881 agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc
     2941 ggtgcttttt tgaattcgct agctaggtct tgaaaggagt gggaattggc tccggtgccc
     3001 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
     3061 attgatccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
     3121 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
     3181 acgttctttt tcgcaacggg tttgccgcca gaacacagga ccggttctag agcgctgcca
     3241 ccatggacaa gaagtacagc atcggcctgg acatcggcac caactctgtg ggctgggccg
     3301 tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc
     3361 ggcacagcat caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg
     3421 aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct
     3481 gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca
     3541 gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg
     3601 gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa
     3661 agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc
     3721 acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg
     3781 acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc
     3841 ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca
     3901 gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa
     3961 acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg
     4021 aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg
     4081 cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca
     4141 tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct
     4201 ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc
     4261 ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg
     4321 ccggctacat tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc
     4381 tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc
     4441 ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc
     4501 acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga
     4561 tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca
     4621 gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg
     4681 aagtggtgga caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata
     4741 agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg
     4801 tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc
     4861 tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga
     4921 ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa
     4981 tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa
     5041 ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg
     5101 tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg
     5161 cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg
     5221 gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc
     5281 tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg
     5341 acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc
     5401 tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga
     5461 cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg
     5521 tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga
     5581 gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc
     5641 ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc
     5701 gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacc
     5761 atatcgtgcc tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa
     5821 gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga
     5881 agaactactg gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc
     5941 tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac
     6001 agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga
     6061 acactaagta cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt
     6121 ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca
     6181 actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa
     6241 agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga
     6301 agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca
     6361 gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc
     6421 ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt
     6481 ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg
     6541 tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc gataagctga
     6601 tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcgacagc cccaccgtgg
     6661 cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg
     6721 tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg
     6781 actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta
     6841 agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct gccggcgaac
     6901 tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca
     6961 gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg
     7021 aacagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag
     7081 tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata
     7141 agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc aatctgggag
     7201 cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagaggtac accagcacca
     7261 aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga
     7321 tcgacctgtc tcagctggga ggcgacaagc gacctgccgc cacaaagaag gctggacagg
     7381 ctaagaagaa gaaagattac aaagacgatg acgataaggg atccggcgca acaaacttct
     7441 ctctgctgaa acaagccgga gatgtcgaag agaatcctgg accgaccgag tacaagccca
     7501 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt
     7561 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca
     7621 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     7681 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     7741 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     7801 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     7861 ccgtcggagt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     7921 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     7981 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     8041 gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg aacgcgttaa gtcgacaatc
     8101 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     8161 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     8221 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     8281 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     8341 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     8401 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     8461 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     8521 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     8581 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     8641 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcgtcga ctttaagacc
     8701 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
     8761 agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg
     8821 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     8881 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     8941 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gggcccgttt
     9001 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     9061 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     9121 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     9181 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     9241 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     9301 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     9361 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     9421 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     9481 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     9541 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     9601 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     9661 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     9721 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     9781 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     9841 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     9901 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     9961 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
    10021 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
    10081 tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc
    10141 ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt
    10201 ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg
    10261 ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg
    10321 ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg
    10381 cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac
    10441 gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg
    10501 cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta
    10561 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
    10621 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
    10681 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
    10741 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
    10801 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
    10861 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
    10921 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
    10981 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
    11041 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
    11101 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
    11161 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
    11221 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
    11281 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
    11341 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
    11401 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
    11461 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
    11521 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
    11581 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
    11641 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
    11701 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
    11761 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
    11821 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
    11881 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
    11941 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
    12001 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
    12061 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
    12121 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
    12181 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
    12241 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
    12301 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
    12361 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
    12421 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
    12481 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
    12541 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
    12601 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
    12661 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
    12721 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
    12781 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
    12841 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
    12901 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
    12961 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cg
//
