LOCUS       sequence-366180-       13013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgRNA targeting human MSH2 gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13013)
  AUTHORS   Povedano JM, Liou J, Wei D, Srivatsav A, Kim J, Xie Y, Nijhawan D, 
            McFadden DG
  TITLE     Engineering Forward Genetics into Cultured Cancer Cells for Chemical
            Target Identification.
  JOURNAL   Cell Chem Biol. 2019 Sep 19;26(9):1315-1321.e3. doi: 
            10.1016/j.chembiol.2019.06.006. Epub 2019 Jul 11.
  PUBMED    31303577
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: 
            "10.1016/j.chembiol.2019.06.006"; journalName: "Cell Chem Biol"; 
            date: "2019-09-19- 19"; volume: "26"; issue: "9"; pages: 
            "1315-1321.e3"
COMMENT     Sequence Label: LentiCRISPR-V2_hMSH2
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(115..135)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        120..449
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      300..435
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     362..381
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        497..544
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             563..937
                     /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    1067..1188
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1104..1123)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1158..1177
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1237..1253)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1237..1253)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1250..1272)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1261..1277
                     /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(1285..1291,1292..1309,1310..1315))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1330..1351
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1468..1485)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1639..2227)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1719..1738)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2398..3189,3190..3258))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPEELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3021..3040
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3259..3363)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3438..3457)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3629..4008
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4010..4208
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4163..4183
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4226..4406
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4453..4578
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5071..5304
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5489..5533
                     /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    5831..5948
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5999..6247
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5999..6019
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6170..6189
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        6269..6344
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        6406..6617
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      6636..6645
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6642..10745
                     /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             6642..10745
                     /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             10746..10793
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             10794..10817
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             10827..10883
                     /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             10884..11480
                     /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     11377..11397
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    11496..12084
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11549..11569)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11967..11978)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             12156..12389
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(12415..12432)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    12421..12645
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(12691..13013,1..106)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12778..12797)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12988..13009
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
       61 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
      121 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
      181 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
      241 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
      301 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
      361 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
      421 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
      481 gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata
      541 cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc
      601 gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt
      661 cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt
      721 ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga
      781 gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc
      841 catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg
      901 caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag atttcgattc
      961 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     1021 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     1081 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     1141 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     1201 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     1261 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     1321 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     1381 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     1441 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1501 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     1561 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1621 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1681 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1741 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1801 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1861 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1921 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1981 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2041 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2101 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2161 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2221 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2281 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2341 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2401 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2461 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     2521 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     2581 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     2641 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     2701 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     2761 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     2821 tagctcctcc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     2881 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     2941 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     3001 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     3061 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     3121 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     3181 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     3241 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     3301 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     3361 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     3421 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     3481 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     3541 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
     3601 cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa
     3661 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     3721 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     3781 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     3841 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     3901 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     3961 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     4021 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     4081 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     4141 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     4201 ataagcagcg cgttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga
     4261 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     4321 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     4381 ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg
     4441 aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc
     4501 gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga
     4561 gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt
     4621 cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg
     4681 gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa
     4741 atactgggac agctacaacc atcccttcag acaggatcag aagaacttag atcattatat
     4801 aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa
     4861 gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc
     4921 gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg aattatataa
     4981 atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt
     5041 ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc
     5101 agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt
     5161 gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct
     5221 gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag
     5281 atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac
     5341 cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca
     5401 cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt
     5461 aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa
     5521 atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt
     5581 cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt
     5641 gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag
     5701 gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc
     5761 cattcgatta gtgaacggat cggcactgcg tgcgccaatt ctgcagacaa atggcagtat
     5821 tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag
     5881 tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc
     5941 aaaattttcg ggtttattac agggacagca gagatccagt ttggttaatt aaggtaccga
     6001 gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg ttagagagat
     6061 aattagaatt aatttgactg taaacacaaa gatattagta caaaatacgt gacgtagaaa
     6121 gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg actatcatat
     6181 gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt ggaaaggacg
     6241 aaacaccgtg agaggctgct taatccacgt tttagagcta gaaatagcaa gttaaaataa
     6301 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt gaattcgcta
     6361 gctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat
     6421 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgatccggt gcctagagaa
     6481 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
     6541 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
     6601 ttgccgccag aacacaggac cggttctaga gcgctgccac catggacaag aagtacagca
     6661 tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg
     6721 tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc
     6781 tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa
     6841 ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca
     6901 gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg
     6961 tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg
     7021 cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg
     7081 acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc
     7141 acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc
     7201 agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg
     7261 acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg
     7321 cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg
     7381 gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga
     7441 gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg
     7501 ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga
     7561 gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg
     7621 agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt
     7681 acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag
     7741 ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg
     7801 aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca
     7861 acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg
     7921 aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc
     7981 gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca
     8041 gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt
     8101 ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg
     8161 tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag
     8221 tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg
     8281 ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg
     8341 actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt
     8401 tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc
     8461 tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg
     8521 aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag
     8581 tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga
     8641 tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg
     8701 gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg
     8761 acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc
     8821 tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc
     8881 tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga
     8941 accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg
     9001 gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc
     9061 agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg
     9121 aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc
     9181 tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga
     9241 gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc
     9301 tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg
     9361 gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga
     9421 tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg
     9481 acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc
     9541 ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg
     9601 cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg
     9661 agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc
     9721 aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg aactttttca
     9781 agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg
     9841 gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc
     9901 tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca
     9961 aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg
    10021 accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg
    10081 ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca
    10141 ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct
    10201 acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg
    10261 aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg
    10321 ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg
    10381 gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg
    10441 acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc
    10501 tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg
    10561 agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact
    10621 ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc
    10681 tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag
    10741 gcgacaagcg acctgccgcc acaaagaagg ctggacaggc taagaagaag aaagattaca
    10801 aagacgatga cgataaggga tccggcgcaa caaacttctc tctgctgaaa caagccggag
    10861 atgtcgaaga gaatcctgga ccgaccgagt acaagcccac ggtgcgcctc gccacccgcg
    10921 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
    10981 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
    11041 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
    11101 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
    11161 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
    11221 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggagtc tcgcccgacc
    11281 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
    11341 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
    11401 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
    11461 cccgcaagcc cggtgcctga acgcgttaag tcgacaatca acctctggat tacaaaattt
    11521 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
    11581 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
    11641 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
    11701 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
    11761 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
    11821 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
    11881 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
    11941 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
    12001 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
    12061 tctccctttg ggccgcctcc ccgcgtcgac tttaagacca atgacttaca aggcagctgt
    12121 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
    12181 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
    12241 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
    12301 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
    12361 cttttagtca gtgtggaaaa tctctagcag ggcccgttta aacccgctga tcagcctcga
    12421 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
    12481 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
    12541 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
    12601 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
    12661 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
    12721 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
    12781 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
    12841 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
    12901 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
    12961 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga aca
//
