LOCUS       sequence-352528-       12797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  nuclease Cas expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12797)
  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 12797)
  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_275
FEATURES             Location/Qualifiers
     source          1..12797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    32..48
                     /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)."
     primer_bind     37..59
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     56..72
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     56..72
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     91..111
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        93..111
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        139..365
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             366..546
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    593..718
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1211..1444
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1629..1673
                     /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    1907..2024
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2163..3341
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2394..3332
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3289..3309
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3358..3378
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3385..7485
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SIRPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASARFLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPRAFKYFDTTIDRKVYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7492..7539
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7552..7605
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             7612..8007
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             8008..8064
                     /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             8071..8769
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     misc_feature    8788..9376
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8841..8861)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9259..9270)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9448..9681
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9759..9880
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9796..9815)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9850..9869
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9920..10055
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(10075..10094)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10076..10094)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10104..10121)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10104..10120)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10113..10135)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10262..10717
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10349..10368)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10559..10580
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10743..10847
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10848..10916,10917..11708)
                     /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     complement(11066..11085)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11879..12467
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12368..12387
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12621..12638
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12755..12776
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12791..12796,12797,1..17,18..24)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
       61 aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag
      121 ctggagctgc aagcttaatg tagtcttatg caatactctt gtagtcttgc aacatggtaa
      181 cgatgagtta gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg
      241 aagtaaggtg gtacgatcgt gccttattag gaaggcaaca gacgggtctg acatggattg
      301 gacgaaccac tgaattgccg cattgcagag atattgtatt taagtgccta gctcgataca
      361 taaacgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
      421 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
      481 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
      541 ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg
      601 acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag
      661 tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag
      721 tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa
      781 gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt
      841 taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc
      901 atcccttcag acaggatcag aagaacttag atcattatat aatacagtag caaccctcta
      961 ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga
     1021 agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag
     1081 gaggagatat gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg
     1141 aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag
     1201 cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg
     1261 cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc
     1321 agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg
     1381 gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc
     1441 tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg
     1501 ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg
     1561 acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc
     1621 agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt
     1681 ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct
     1741 tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat
     1801 attcaccatt atcgtttcag acccacctcc caaccccgag gggaccctgg tacccatgca
     1861 tgcatgaatt ctagatcttg agacaaatgg cagtattcat ccacaatttt aaaagaaaag
     1921 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     1981 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2041 acagcagaga tccactttgg cgccggctcg agggggcccg ggtgcaaaga tggataaagt
     2101 tttaaacaga gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa
     2161 ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
     2221 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
     2281 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     2341 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
     2401 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
     2461 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
     2521 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
     2581 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
     2641 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
     2701 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
     2761 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
     2821 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
     2881 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
     2941 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
     3001 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
     3061 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
     3121 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
     3181 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
     3241 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
     3301 cagtggttca aagttttttt cttccatttc aggtgtcgtg agggatccac catggcacca
     3361 aagaagaagc ggaaggtcgc agccgacaag aagtacagca tcggcctgga catcggcacc
     3421 aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag
     3481 gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc
     3541 gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc
     3601 agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg
     3661 gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac
     3721 gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc
     3781 accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg
     3841 atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac
     3901 ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac
     3961 cagctgttcg aggaaaaccc catcaacgcc agcggcgtag acgccaaggc catcctgtct
     4021 gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag
     4081 aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag
     4141 agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac
     4201 gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc
     4261 aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc
     4321 aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc
     4381 ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac
     4441 cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga agagttctac
     4501 aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg
     4561 aacagagagg acctgctgcg gaagcagcgt acgttcgaca acggcagcat cccccaccag
     4621 atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg
     4681 aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc
     4741 cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc
     4801 accccctgga acttcgagga agtggtggac aagggcgcta gcgcccagag cttcatcgag
     4861 cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg
     4921 ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga
     4981 atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc
     5041 aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag
     5101 tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca
     5161 taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag
     5221 gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag
     5281 gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg
     5341 cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag
     5401 cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc
     5461 atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg
     5521 tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt
     5581 aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg
     5641 cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga
     5701 cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc
     5761 cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg
     5821 tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg
     5881 tccgactacg atgtggacca tatcgtgcct cagagctttc tgaaggacga ctccatcgac
     5941 aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa
     6001 gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc
     6061 cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag
     6121 gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag
     6181 atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg
     6241 aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac
     6301 aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg
     6361 ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac
     6421 aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc
     6481 gccaagtact tcttctacag caacatcatg aactttttca agaccgagat taccctggcc
     6541 aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg
     6601 tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat
     6661 atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat ccggcccaag
     6721 aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc
     6781 ttcgtgagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag
     6841 tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc
     6901 ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac
     6961 ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg
     7021 ctggcctctg cccggttcct gcagaaggga aacgaactgg ccctgccctc caaatatgtg
     7081 aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag
     7141 cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc
     7201 agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc
     7261 tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt
     7321 accctgacca atctgggagc ccctcgggcc ttcaagtact ttgacaccac catcgaccgg
     7381 aaggtgtacc ggagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc
     7441 ggcctgtacg agacacgaat cgatttgtct cagctgggag gcgacgccgc aaaaaggccg
     7501 gcggccacga aaaaggccgg ccaggcaaaa aagaaaaagg caggctcagg cgagggcaga
     7561 ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg gcccaagcgc tatggccaag
     7621 cctttgtctc aagaagaatc caccctcatt gaaagagcaa cggctacaat caacagcatc
     7681 cccatctctg aagactacag cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc
     7741 actggtgtca atgtatatca ttttactggg ggaccttgtg cagaactcgt ggtgctgggc
     7801 actgctgctg ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac
     7861 aggggcatct tgagcccctg cggacggtgc cgacaggtgc ttctcgatct gcatcctggg
     7921 atcaaagcca tagtgaagga cagtgatgga cagccgacgg cagttgggat tcgtgaattg
     7981 ctgccctctg gttatgtgtg ggagggcgca acaaacttct ctctgctgaa acaagccgga
     8041 gatgtcgaag agaatcctgg accgactagt atggtgagcg agctgattaa ggagaacatg
     8101 cacatgaagc tatacatgga gggcaccgtg aacaaccacc acttcaagtg cacatccgag
     8161 ggcgaaggca agccctacga gggcacccag accatgagaa tcaaggcggt cgagggcggc
     8221 cctctcccct tcgccttcga catcctggct accagcttca tgtacggcag caaaaccttc
     8281 atcaaccaca cccagggcat ccccgacttc tttaagcagt ccttccccga gggcttcaca
     8341 tgggagagag tcaccacata cgaagacggg ggcgtgctga ccgctaccca ggacaccagc
     8401 ctccaggacg gctgcctcat ctacaacgtc aagatcagag gggtgaactt cccatccaac
     8461 ggccctgtga tgcagaagaa aacactcggc tgggaggcct ccaccgagac cctgtacccc
     8521 gctgacggcg gcctggaagg cagagccgac atggccctga agctcgtggg cgggggccac
     8581 ctgatctgca acttgaagac cacatacaga tccaagaaac ccgctaagaa cctcaagatg
     8641 cccggcgtct actatgtgga cagaagactg gaaagaatca aggaggccga caaagagacc
     8701 tacgtcgagc agcacgaggt ggctgtggcc agatactgcg acctccctag caaactgggg
     8761 cacagatgat agacgcgtta agtcgacaat caacctctgg attacaaaat ttgtgaaaga
     8821 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     8881 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     8941 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     9001 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     9061 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     9121 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     9181 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     9241 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     9301 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     9361 tgggccgcct ccccgcgtcg actttaagac caatgactta caaggcagct gtagatctta
     9421 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag
     9481 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     9541 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     9601 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     9661 cagtgtggaa aatctctagc agtacgtata gtagttcatg tcatcttatt attcagtatt
     9721 tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata
     9781 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     9841 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat
     9901 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     9961 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
    10021 attccagaag tagtgaggag gcttttttgg aggcctaggg acgtacccaa ttcgccctat
    10081 agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac
    10141 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
    10201 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
    10261 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
    10321 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
    10381 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
    10441 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
    10501 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
    10561 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
    10621 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
    10681 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
    10741 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
    10801 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
    10861 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
    10921 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
    10981 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
    11041 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
    11101 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
    11161 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
    11221 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
    11281 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
    11341 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
    11401 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
    11461 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
    11521 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
    11581 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
    11641 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
    11701 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
    11761 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
    11821 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
    11881 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
    11941 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
    12001 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
    12061 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
    12121 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
    12181 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
    12241 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
    12301 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
    12361 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
    12421 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
    12481 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
    12541 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
    12601 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
    12661 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
    12721 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
    12781 caccccaggc tttacac
//
