LOCUS       sequence-352524-       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_154
FEATURES             Location/Qualifiers
     source          1..12797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..170
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     404..421
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    538..559
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(574..579,580..597,598..604)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    612..628
                     /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     617..639
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     636..652
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     636..652
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     671..691
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        673..691
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        719..945
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             946..1126
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1173..1298
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1791..2024
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2209..2253
                     /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    2487..2604
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2743..3921
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2974..3912
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3869..3889
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3938..3958
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3965..8065
                     /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
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDTKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKLYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGIIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEKVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGDQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFIQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGVLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             8072..8119
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8132..8185
                     /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             8192..8587
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             8588..8644
                     /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             8651..9349
                     /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    9368..9956
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9421..9441)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9839..9850)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10028..10261
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    10339..10460
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10376..10395)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10430..10449
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      10500..10635
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(10655..10674)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10656..10674)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10684..10701)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10684..10700)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10693..10715)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10842..11297
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10929..10948)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11139..11160
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        11323..11427
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11428..11496,11497..12288)
                     /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(11646..11665)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(12459..12797,1..250)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
       61 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
      121 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
      181 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
      241 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
      301 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
      361 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
      421 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
      481 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
      541 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
      601 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
      661 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg
      721 tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc
      781 ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt
      841 gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg
      901 cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta
      961 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     1021 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     1081 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
     1141 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
     1201 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     1261 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     1321 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     1381 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
     1441 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     1501 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     1561 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     1621 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat
     1681 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     1741 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     1801 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     1861 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     1921 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     1981 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     2041 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     2101 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
     2161 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
     2221 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
     2281 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
     2341 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
     2401 acccacctcc caaccccgag gggaccctgg tacccatgca tgcatgaatt ctagatcttg
     2461 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     2521 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     2581 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccactttgg
     2641 cgccggctcg agggggcccg ggtgcaaaga tggataaagt tttaaacaga gaggaatctt
     2701 tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa ttggctccgg tgcccgtcag
     2761 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
     2821 accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
     2881 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
     2941 ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg
     3001 cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac ctggctgcag
     3061 tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt gggagagttc gaggccttgc
     3121 gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg ctggggccgc
     3181 cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc
     3241 atttaaaatt tttgatgacc tgctgcgacg ctttttttct ggcaagatag tcttgtaaat
     3301 gcgggccaag atctgcacac tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc
     3361 cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc accgagaatc
     3421 ggacgggggt agtctcaagc tggccggcct gctctggtgc ctggcctcgc gccgccgtgt
     3481 atcgccccgc cctgggcggc aaggctggcc cggtcggcac cagttgcgtg agcggaaaga
     3541 tggccgcttc ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag
     3601 cgggcgggtg agtcacccac acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca
     3661 tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg attagttctc gagcttttgg
     3721 agtacgtcgt ctttaggttg gggggagggg ttttatgcga tggagtttcc ccacactgag
     3781 tgggtggaga ctgaagttag gccagcttgg cacttgatgt aattctcctt ggaatttgcc
     3841 ctttttgagt ttggatcttg gttcattctc aagcctcaga cagtggttca aagttttttt
     3901 cttccatttc aggtgtcgtg agggatccac catggcacca aagaagaagc ggaaggtcgc
     3961 agccgacaag aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt
     4021 gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg
     4081 gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga
     4141 ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg
     4201 ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag
     4261 actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg
     4321 caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa
     4381 gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca
     4441 catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga
     4501 cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc
     4561 catcaacgcc agcggcgtag acgccaaggc catcctgtct gccagactga gcaagagcag
     4621 acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa
     4681 cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga
     4741 ggataccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc
     4801 ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat
     4861 cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc
     4921 tatgatcaag ctgtacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg
     4981 gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc
     5041 cggctacatt gacggcggag ccagccagga agagttctac aagttcatca agcccatcct
     5101 ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg
     5161 gaagcagcgt acgttcgaca acggcatcat cccccaccag atccacctgg gagagctgca
     5221 cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat
     5281 cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag
     5341 cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagaa
     5401 agtggtggac aagggcgcta gcgcccagag cttcatcgag cggatgacca acttcgataa
     5461 gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt
     5521 gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct
     5581 gagcggcgac cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac
     5641 cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat
     5701 ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat
     5761 tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt
     5821 gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc
     5881 ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg
     5941 caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct
     6001 ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atccagctga tccacgacga
     6061 cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct
     6121 gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac
     6181 agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt
     6241 gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag
     6301 aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc
     6361 cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg
     6421 ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg atgtggacca
     6481 tatcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag
     6541 cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa
     6601 gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct
     6661 gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca
     6721 gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa
     6781 cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc
     6841 caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa
     6901 ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa
     6961 gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa
     7021 gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag
     7081 caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg
     7141 gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt
     7201 tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt
     7261 gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat
     7321 cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc
     7381 ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt
     7441 gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga
     7501 ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa
     7561 gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgtgct
     7621 gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag
     7681 ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga
     7741 acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt
     7801 gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa
     7861 gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc
     7921 ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa
     7981 agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacgaat
     8041 cgatttgtct cagctgggag gcgacgccgc aaaaaggccg gcggccacga aaaaggccgg
     8101 ccaggcaaaa aagaaaaagg caggctcagg cgagggcaga ggaagtctgc taacatgcgg
     8161 tgacgtcgag gagaatcctg gcccaagcgc tatggccaag cctttgtctc aagaagaatc
     8221 caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg aagactacag
     8281 cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca atgtatatca
     8341 ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg ctgcggcagc
     8401 tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct tgagcccctg
     8461 cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca tagtgaagga
     8521 cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg gttatgtgtg
     8581 ggagggcgca acaaacttct ctctgctgaa acaagccgga gatgtcgaag agaatcctgg
     8641 accgactagt atggtgagcg agctgattaa ggagaacatg cacatgaagc tatacatgga
     8701 gggcaccgtg aacaaccacc acttcaagtg cacatccgag ggcgaaggca agccctacga
     8761 gggcacccag accatgagaa tcaaggcggt cgagggcggc cctctcccct tcgccttcga
     8821 catcctggct accagcttca tgtacggcag caaaaccttc atcaaccaca cccagggcat
     8881 ccccgacttc tttaagcagt ccttccccga gggcttcaca tgggagagag tcaccacata
     8941 cgaagacggg ggcgtgctga ccgctaccca ggacaccagc ctccaggacg gctgcctcat
     9001 ctacaacgtc aagatcagag gggtgaactt cccatccaac ggccctgtga tgcagaagaa
     9061 aacactcggc tgggaggcct ccaccgagac cctgtacccc gctgacggcg gcctggaagg
     9121 cagagccgac atggccctga agctcgtggg cgggggccac ctgatctgca acttgaagac
     9181 cacatacaga tccaagaaac ccgctaagaa cctcaagatg cccggcgtct actatgtgga
     9241 cagaagactg gaaagaatca aggaggccga caaagagacc tacgtcgagc agcacgaggt
     9301 ggctgtggcc agatactgcg acctccctag caaactgggg cacagatgat agacgcgtta
     9361 agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     9421 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     9481 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     9541 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     9601 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     9661 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     9721 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     9781 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     9841 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     9901 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgtcg
     9961 actttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
    10021 gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact
    10081 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
    10141 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
    10201 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
    10261 agtacgtata gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
    10321 atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat
    10381 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
    10441 aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca
    10501 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
    10561 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
    10621 gcttttttgg aggcctaggg acgtacccaa ttcgccctat agtgagtcgt attacgcgcg
    10681 ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
    10741 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
    10801 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
    10861 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
    10921 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
    10981 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
    11041 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
    11101 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
    11161 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
    11221 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
    11281 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
    11341 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
    11401 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
    11461 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
    11521 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
    11581 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
    11641 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
    11701 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
    11761 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
    11821 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
    11881 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
    11941 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
    12001 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
    12061 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
    12121 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
    12181 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
    12241 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
    12301 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
    12361 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
    12421 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
    12481 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
    12541 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
    12601 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
    12661 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
    12721 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
    12781 gggggttcgt gcacaca
//
