LOCUS       sequence-352548-       13663 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BE Cas expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13663)
  AUTHORS   Sangree AK, Griffith AL, Szegletes ZM, Roy P, DeWeirdt PC, Hegde M, 
            McGee AV, Hanna RE, Doench JG
  TITLE     Benchmarking of SpCas9 variants enables deeper base editor screens 
            of BRCA1 and BCL2.
  JOURNAL   Nat Commun. 2022 Mar 14;13(1):1318. doi: 10.1038/s41467-022-28884-7.
  PUBMED    35288574
REFERENCE   2  (bases 1 to 13663)
  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: "Nat 
            Commun."; date: "2022-03-14"; volume: "13(1):1318. doi"; pages: " 
            10.1038/s41467-022-28884-7"
COMMENT     Sequence Label: pRDA_426
FEATURES             Location/Qualifiers
     source          1..13663
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     42..61
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        177..224
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             243..617
                     /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    747..868
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(784..803)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     838..857
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(917..933)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(917..933)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(930..952)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    941..957
                     /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(965..971,972..989,990..995))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1010..1031
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1148..1165)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1319..1907)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1399..1418)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2078..2869,2870..2938))
                     /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     2701..2720
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2939..3043)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3118..3137)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3309..3688
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3690..3888
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3843..3863
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             3906..4086
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4133..4258
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4751..4984
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5169..5213
                     /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    5511..5628
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5679..5927
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5679..5699
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     5850..5869
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        5949..6024
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        6108..6319
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             6369..6389
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(6984..7007,7008..7010,7011..11084)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6984..11084
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             11127..11147
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11157..11213
                     /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             11214..11810
                     /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     11707..11727
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    11826..12414
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11879..11899)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(12297..12308)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             12486..12719
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(12745..12762)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    12751..12975
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      13021..13449
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(13108..13127)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     13318..13339
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(13458..13478)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(13463..13663,1..129)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(13643..13663,1..115)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
       61 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
      121 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg
      181 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
      241 ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg
      301 tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg
      361 gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg
      421 acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg
      481 aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc
      541 agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg
      601 ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa
      661 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
      721 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
      781 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
      841 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
      901 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
      961 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     1021 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     1081 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     1141 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     1201 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1261 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1321 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1381 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1441 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1501 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     1561 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     1621 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     1681 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     1741 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     1801 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     1861 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     1921 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     1981 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2041 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2101 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2161 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     2221 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2281 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     2341 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     2401 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     2461 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     2521 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     2581 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     2641 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     2701 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     2761 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     2821 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     2881 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     2941 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3001 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     3061 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
     3121 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
     3181 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
     3241 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
     3301 tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     3361 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     3421 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     3481 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     3541 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     3601 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     3661 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
     3721 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     3781 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
     3841 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagcg cgttttgcct
     3901 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     3961 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     4021 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     4081 ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg
     4141 acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag
     4201 tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag
     4261 tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa
     4321 gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt
     4381 taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc
     4441 atcccttcag acaggatcag aagaacttag atcattatat aatacagtag caaccctcta
     4501 ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga
     4561 agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag
     4621 gaggagatat gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg
     4681 aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag
     4741 cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg
     4801 cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc
     4861 agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg
     4921 gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc
     4981 tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg
     5041 ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg
     5101 acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc
     5161 agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt
     5221 ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct
     5281 tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat
     5341 attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag
     5401 gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat
     5461 cggcactgcg tgcgccaatt ctgcagacaa atggcagtat tcatccacaa ttttaaaaga
     5521 aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac
     5581 atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac
     5641 agggacagca gagatccagt ttggttaatt aaggtaccga gggcctattt cccatgattc
     5701 cttcatattt gcatatacga tacaaggctg ttagagagat aattagaatt aatttgactg
     5761 taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt
     5821 ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt
     5881 atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacaccgga gacgcttttt
     5941 tcgtctcagt ttgagagcta gaaatagcaa gttcaaataa ggctagtccg ttatcaactt
     6001 gaaaaagtgg caccgagtcg gtgcgctcac ctattagcgg ctaaggtttt ttgaattcgc
     6061 tagctaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg cagagcgcac
     6121 atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgatccg gtgcctagag
     6181 aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga
     6241 gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg
     6301 gtttgccgcc agaacacagg accggtgcca ccatgaaacg gacagccgac ggaagcgagt
     6361 tcgagtcacc aaagaagaag cggaaagtct ctgaggtgga gttttcccac gagtactgga
     6421 tgagacatgc cctgaccctg gccaagaggg cacgggatga gagggaggtg cctgtgggag
     6481 ccgtgctggt gctgaacaat agagtgatcg gcgagggctg gaacagagcc atcggcctgc
     6541 acgacccaac agcccatgcc gaaattatgg ccctgagaca gggcggcctg gtcatgcaga
     6601 actacagact gattgacgcc accctgtacg tgacattcga gccttgcgtg atgtgcgccg
     6661 gcgccatgat ccactctagg atcggccgcg tggtgtttgg atggagaaat tctaaaagag
     6721 gcgccgcagg ctccctgatg aacgtgctga actaccccgg catgaatcac cgcgtcgaaa
     6781 ttaccgaggg aatcctggca gatgaatgtg ccgccctgct gtgcgatttc tatcggatgc
     6841 ctagacaggt gttcaatgct cagaagaagg cccagagctc catcaactct ggaggatcta
     6901 gcggaggatc ctctggcagc gagacaccag gaacaagcga gtcagcaaca ccagagagca
     6961 gtggcggcag cagcggcggc agcgacaaga agtacagcat cggcctggcc atcggcacca
     7021 actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg
     7081 tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg
     7141 acagcggcga aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca
     7201 gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg
     7261 acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg
     7321 agcggcaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca
     7381 ccatctacca cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga
     7441 tctatctggc cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc
     7501 tgaaccccga caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc
     7561 agctgttcga ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg
     7621 ccagactgag caagagcaga cggctggaaa atctgatcgc ccagctgccc ggcgagaaga
     7681 agaatggcct gttcggaaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga
     7741 gcaacttcga cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg
     7801 acctggacaa cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca
     7861 agaacctgtc cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca
     7921 aggcccccct gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc
     7981 tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc
     8041 agagcaagaa cggctacgcc ggctacattg acggcggagc cagccaggaa gagttctaca
     8101 agttcatcaa gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga
     8161 acagagagga cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga
     8221 tccacctggg agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga
     8281 aggacaaccg ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc
     8341 ctctggccag gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca
     8401 ccccctggaa cttcgaggaa gtggtggaca agggcgcttc cgcccagagc ttcatcgagc
     8461 ggatgaccaa cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc
     8521 tgtacgagta cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg accgagggaa
     8581 tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc catcgtggac ctgctgttca
     8641 agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt
     8701 gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat
     8761 accacgatct gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg
     8821 acattctgga agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg
     8881 aacggctgaa aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc
     8941 ggagatacac cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc
     9001 agtccggcaa gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca
     9061 tgcagctgat ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt
     9121 ccggccaggg cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta
     9181 agaagggcat cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc
     9241 acaagcccga gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac
     9301 agaagaacag ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc
     9361 agatcctgaa agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt
     9421 actacctgca gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt
     9481 ccgactacga tgtggaccat atcgtgcctc agagctttct gaaggacgac tccatcgaca
     9541 acaaggtgct gaccagaagc gacaagaacc ggggcaagag cgacaacgtg ccctccgaag
     9601 aggtcgtgaa gaagatgaag aactactggc ggcagctgct gaacgccaag ctgattaccc
     9661 agagaaagtt cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg
     9721 ccggcttcat caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga
     9781 tcctggactc ccggatgaac actaagtacg acgagaatga caagctgatc cgggaagtga
     9841 aagtgatcac cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca
     9901 aagtgcgcga gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg
     9961 gaaccgccct gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca
    10021 aggtgtacga cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg
    10081 ccaagtactt cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca
    10141 acggcgagat ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt
    10201 gggataaggg ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc caagtgaata
    10261 tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga
    10321 ggaacagcga taagctgatc gccagaaaga aggactggga ccctaagaag tacggcggct
    10381 tcgacagccc caccgtggcc tattctgtgc tggtggtggc caaagtggaa aagggcaagt
    10441 ccaagaaact gaagagtgtg aaagagctgc tggggatcac catcatggaa agaagcagct
    10501 tcgagaagaa tcccatcgac tttctggaag ccaagggcta caaagaagtg aaaaaggacc
    10561 tgatcatcaa gctgcctaag tactccctgt tcgagctgga aaacggccgg aagagaatgc
    10621 tggcctctgc cggcgaactg cagaagggaa acgaactggc cctgccctcc aaatatgtga
    10681 acttcctgta cctggccagc cactatgaga agctgaaggg ctcccccgag gataatgagc
    10741 agaaacagct gtttgtggaa cagcacaagc actacctgga cgagatcatc gagcagatca
    10801 gcgagttctc caagagagtg atcctggccg acgctaatct ggacaaagtg ctgtccgcct
    10861 acaacaagca ccgggataag cccatcagag agcaggccga gaatatcatc cacctgttta
    10921 ccctgaccaa tctgggagcc cctgccgcct tcaagtactt tgacaccacc atcgaccgga
    10981 agaggtacac cagcaccaaa gaggtgctgg acgccaccct gatccaccag agcatcaccg
    11041 gcctgtacga gacacggatc gacctgtctc agctgggagg tgacagcgga ggctccaaaa
    11101 gaaccgccga cggcagcgag ttcgagccca agaagaagag gaaagtcagc gctggcgcaa
    11161 caaacttctc tctgctgaaa caagccggag atgtcgaaga gaatcctgga ccgaccgagt
    11221 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg
    11281 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
    11341 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
    11401 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
    11461 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
    11521 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
    11581 tcctggccac cgtcggagtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
    11641 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
    11701 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
    11761 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga acgcgttaag
    11821 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
    11881 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
    11941 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
    12001 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
    12061 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
    12121 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
    12181 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
    12241 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
    12301 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
    12361 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcgtcgac
    12421 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg
    12481 gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg
    12541 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
    12601 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
    12661 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
    12721 ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
    12781 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
    12841 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
    12901 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
    12961 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
    13021 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
    13081 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
    13141 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
    13201 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
    13261 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
    13321 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
    13381 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
    13441 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
    13501 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
    13561 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
    13621 agtcccgccc ctaactccgc ccatcccgcc cctaactccg ccc
//
