LOCUS       sequence-352549-       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_429
FEATURES             Location/Qualifiers
     source          1..13663
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..103
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             122..496
                     /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    626..747
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(663..682)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     717..736
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(796..812)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(796..812)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(809..831)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    820..836
                     /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(844..850,851..868,869..874))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    889..910
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1027..1044)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1198..1786)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1278..1297)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1957..2748,2749..2817))
                     /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     2580..2599
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2818..2922)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2997..3016)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3188..3567
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3569..3767
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3722..3742
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             3785..3965
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4012..4137
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4630..4863
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5048..5092
                     /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    5390..5507
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5558..5806
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5558..5578
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     5729..5748
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        5828..5903
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        5987..6198
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             6248..6268
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6863..10963
                     /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
                     SIRPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASARFLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPRAFKYFDTTIDRKVYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             11006..11026
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11036..11092
                     /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             11093..11689
                     /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     11586..11606
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    11705..12293
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11758..11778)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(12176..12187)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             12365..12598
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(12624..12641)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    12630..12854
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      12900..13328
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12987..13006)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     13197..13218
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(13337..13357)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(13342..13663,1..8)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      13522..13657
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     13584..13603
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga
       61 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
      121 catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt
      181 cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg
      241 tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga
      301 caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga
      361 ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca
      421 gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc
      481 cgaggagcag gactgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag
      541 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
      601 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
      661 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
      721 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
      781 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
      841 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
      901 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
      961 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     1021 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     1081 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     1141 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1201 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1261 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1321 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1381 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1441 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     1501 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     1561 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     1621 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     1681 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     1741 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     1801 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     1861 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     1921 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     1981 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     2041 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     2101 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     2161 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     2221 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     2281 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     2341 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     2401 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     2461 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     2521 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     2581 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     2641 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     2701 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     2761 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     2821 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     2881 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     2941 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
     3001 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     3061 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     3121 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
     3181 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
     3241 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     3301 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     3361 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     3421 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     3481 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     3541 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     3601 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     3661 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     3721 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg
     3781 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     3841 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     3901 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     3961 tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga
     4021 cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt
     4081 acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt
     4141 attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag
     4201 aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt
     4261 aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca
     4321 tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc aaccctctat
     4381 tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa
     4441 gagcaaaaca aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg
     4501 aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga
     4561 accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc
     4621 agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc
     4681 agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca
     4741 gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg
     4801 catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct
     4861 cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc
     4921 tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga
     4981 cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca
     5041 gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg
     5101 gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt
     5161 ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata
     5221 ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg
     5281 aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc
     5341 ggcactgcgt gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa
     5401 aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca
     5461 tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca
     5521 gggacagcag agatccagtt tggttaatta aggtaccgag ggcctatttc ccatgattcc
     5581 ttcatatttg catatacgat acaaggctgt tagagagata attagaatta atttgactgt
     5641 aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt
     5701 tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta
     5761 tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccggag acgctttttt
     5821 cgtctcagtt tgagagctag aaatagcaag ttcaaataag gctagtccgt tatcaacttg
     5881 aaaaagtggc accgagtcgg tgcgctcacc tattagcggc taaggttttt tgaattcgct
     5941 agctaggtct tgaaaggagt gggaattggc tccggtgccc gtcagtgggc agagcgcaca
     6001 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgatccgg tgcctagaga
     6061 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     6121 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     6181 tttgccgcca gaacacagga ccggtgccac catgaaacgg acagccgacg gaagcgagtt
     6241 cgagtcacca aagaagaagc ggaaagtctc tgaggtggag ttttcccacg agtactggat
     6301 gagacatgcc ctgaccctgg ccaagagggc acgggatgag agggaggtgc ctgtgggagc
     6361 cgtgctggtg ctgaacaata gagtgatcgg cgagggctgg aacagagcca tcggcctgca
     6421 cgacccaaca gcccatgccg aaattatggc cctgagacag ggcggcctgg tcatgcagaa
     6481 ctacagactg attgacgcca ccctgtacgt gacattcgag ccttgcgtga tgtgcgccgg
     6541 cgccatgatc cactctagga tcggccgcgt ggtgtttgga tggagaaatt ctaaaagagg
     6601 cgccgcaggc tccctgatga acgtgctgaa ctaccccggc atgaatcacc gcgtcgaaat
     6661 taccgaggga atcctggcag atgaatgtgc cgccctgctg tgcgatttct atcggatgcc
     6721 tagacaggtg ttcaatgctc agaagaaggc ccagagctcc atcaactctg gaggatctag
     6781 cggaggatcc tctggcagcg agacaccagg aacaagcgag tcagcaacac cagagagcag
     6841 tggcggcagc agcggcggca gcgacaagaa gtacagcatc ggcctggcca tcggcaccaa
     6901 ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt
     6961 gctgggcaac accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga
     7021 cagcggcgaa acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag
     7081 acggaagaac cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga
     7141 cgacagcttc ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga
     7201 gcggcacccc atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac
     7261 catctaccac ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat
     7321 ctatctggcc ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct
     7381 gaaccccgac aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca
     7441 gctgttcgag gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc
     7501 cagactgagc aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa
     7561 gaatggcctg ttcggaaacc tgattgccct gagcctgggc ctgaccccca acttcaagag
     7621 caacttcgac ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga
     7681 cctggacaac ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa
     7741 gaacctgtcc gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa
     7801 ggcccccctg agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct
     7861 gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca
     7921 gagcaagaac ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa
     7981 gttcatcaag cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa
     8041 cagagaggac ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat
     8101 ccacctggga gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa
     8161 ggacaaccgg gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc
     8221 tctggccagg ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac
     8281 cccctggaac ttcgaggaag tggtggacaa gggcgcttcc gcccagagct tcatcgagcg
     8341 gatgaccaac ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct
     8401 gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat
     8461 gagaaagccc gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa
     8521 gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg
     8581 cttcgactcc gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata
     8641 ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga
     8701 cattctggaa gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga
     8761 acggctgaaa acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg
     8821 gagatacacc ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca
     8881 gtccggcaag acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat
     8941 gcagctgatc cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc
     9001 cggccagggc gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa
     9061 gaagggcatc ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca
     9121 caagcccgag aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca
     9181 gaagaacagc cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca
     9241 gatcctgaaa gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta
     9301 ctacctgcag aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc
     9361 cgactacgat gtggaccata tcgtgcctca gagctttctg aaggacgact ccatcgacaa
     9421 caaggtgctg accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga
     9481 ggtcgtgaag aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca
     9541 gagaaagttc gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc
     9601 cggcttcatc aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat
     9661 cctggactcc cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa
     9721 agtgatcacc ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa
     9781 agtgcgcgag atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg
     9841 aaccgccctg atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa
     9901 ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc
     9961 caagtacttc ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa
    10021 cggcgagatc cggaagcggc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg
    10081 ggataagggc cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat
    10141 cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa gagtctatcc ggcccaagag
    10201 gaacagcgat aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt
    10261 cgtgagcccc accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc
    10321 caagaaactg aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt
    10381 cgagaagaat cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct
    10441 gatcatcaag ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct
    10501 ggcctctgcc cggttcctgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa
    10561 cttcctgtac ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca
    10621 gaaacagctg tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag
    10681 cgagttctcc aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta
    10741 caacaagcac cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac
    10801 cctgaccaat ctgggagccc ctcgggcctt caagtacttt gacaccacca tcgaccggaa
    10861 ggtgtaccgg agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg
    10921 cctgtacgag acacggatcg acctgtctca gctgggaggt gacagcggag gctccaaaag
    10981 aaccgccgac ggcagcgagt tcgagcccaa gaagaagagg aaagtcagcg ctggcgcaac
    11041 aaacttctct ctgctgaaac aagccggaga tgtcgaagag aatcctggac cgaccgagta
    11101 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc
    11161 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga
    11221 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
    11281 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
    11341 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
    11401 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
    11461 cctggccacc gtcggagtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
    11521 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
    11581 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
    11641 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgaa cgcgttaagt
    11701 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
    11761 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
    11821 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
    11881 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
    11941 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
    12001 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
    12061 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
    12121 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
    12181 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
    12241 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcgtcgact
    12301 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
    12361 ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg
    12421 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
    12481 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
    12541 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagg
    12601 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
    12661 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
    12721 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
    12781 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
    12841 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
    12901 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
    12961 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
    13021 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
    13081 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
    13141 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
    13201 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
    13261 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
    13321 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
    13381 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
    13441 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
    13501 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
    13561 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
    13621 ctattccaga agtagtgagg aggctttttt ggaggcctag gct
//
