LOCUS       sequence-352521-       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_151
FEATURES             Location/Qualifiers
     source          1..12797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    93..114
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(129..134,135..152,153..159)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    167..183
                     /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     172..194
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     191..207
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     191..207
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     226..246
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        228..246
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        274..500
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             501..681
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    728..853
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1346..1579
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1764..1808
                     /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    2042..2159
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2298..3476
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2529..3467
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3424..3444
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3493..3513
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3520..7620
                     /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
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTGFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGALSRKLINGIRDKQSGKTILDFLKSDGFANRNFMA
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRAITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7627..7674
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7687..7740
                     /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             7747..8142
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             8143..8199
                     /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             8206..8904
                     /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    8923..9511
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8976..8996)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9394..9405)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9583..9816
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9894..10015
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9931..9950)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9985..10004
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      10055..10190
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(10210..10229)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10211..10229)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10239..10256)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10239..10255)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10248..10270)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10397..10852
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10484..10503)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10694..10715
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10878..10982
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10983..11051,11052..11843)
                     /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(11201..11220)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12014..12602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12503..12522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12756..12773
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
       61 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
      121 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
      181 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac
      241 taaagggaac aaaagctgga gctgcaagct taatgtagtc ttatgcaata ctcttgtagt
      301 cttgcaacat ggtaacgatg agttagcaac atgccttaca aggagagaaa aagcaccgtg
      361 catgccgatt ggtggaagta aggtggtacg atcgtgcctt attaggaagg caacagacgg
      421 gtctgacatg gattggacga accactgaat tgccgcattg cagagatatt gtatttaagt
      481 gcctagctcg atacataaac gggtctctct ggttagacca gatctgagcc tgggagctct
      541 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
      601 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
      661 cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc
      721 agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg
      781 gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg
      841 gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt
      901 aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct
      961 agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact
     1021 gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac
     1081 agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt
     1141 agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga
     1201 tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata
     1261 aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc
     1321 agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag
     1381 gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg
     1441 gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc
     1501 aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc
     1561 taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg
     1621 ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga
     1681 cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg
     1741 aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg
     1801 caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa
     1861 tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata
     1921 gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac
     1981 cctggtaccc atgcatgcat gaattctaga tcttgagaca aatggcagta ttcatccaca
     2041 attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa
     2101 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc
     2161 gggtttatta cagggacagc agagatccac tttggcgccg gctcgagggg gcccgggtgc
     2221 aaagatggat aaagttttaa acagagagga atctttgcag ctaatggacc ttctaggtct
     2281 tgaaaggagt gggaattggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag
     2341 tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg
     2401 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
     2461 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca
     2521 gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc
     2581 cttgcgtgcc ttgaattact tccacctggc tgcagtacgt gattcttgat cccgagcttc
     2641 gggttggaag tgggtgggag agttcgaggc cttgcgctta aggagcccct tcgcctcgtg
     2701 cttgagttga ggcctggcct gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc
     2761 gcgcctgtct cgctgctttc gataagtctc tagccattta aaatttttga tgacctgctg
     2821 cgacgctttt tttctggcaa gatagtcttg taaatgcggg ccaagatctg cacactggta
     2881 tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg
     2941 cgaggcgggg cctgcgagcg cggccaccga gaatcggacg ggggtagtct caagctggcc
     3001 ggcctgctct ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc
     3061 tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag
     3121 ggagctcaaa atggaggacg cggcgctcgg gagagcgggc gggtgagtca cccacacaaa
     3181 ggaaaagggc ctttccgtcc tcagccgtcg cttcatgtga ctccacggag taccgggcgc
     3241 cgtccaggca cctcgattag ttctcgagct tttggagtac gtcgtcttta ggttgggggg
     3301 aggggtttta tgcgatggag tttccccaca ctgagtgggt ggagactgaa gttaggccag
     3361 cttggcactt gatgtaattc tccttggaat ttgccctttt tgagtttgga tcttggttca
     3421 ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc atttcaggtg tcgtgaggga
     3481 tccaccatgg caccaaagaa gaagcggaag gtcgcagccg acaagaagta cagcatcggc
     3541 ctggacatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc
     3601 agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc
     3661 ggagccctgc tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc
     3721 agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac
     3781 gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa
     3841 gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac
     3901 cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag
     3961 gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc
     4021 ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg
     4081 gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtagacgcc
     4141 aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag
     4201 ctgcccggcg agaagaagaa tggcctgttc ggaaacctga ttgccctgag cctgggcctg
     4261 acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag
     4321 gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac
     4381 ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg
     4441 aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac
     4501 caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa
     4561 gagattttct tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc
     4621 caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa
     4681 ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcgtacgtt cgacaacggc
     4741 agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat
     4801 ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc
     4861 ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag
     4921 agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgctagcgcc
     4981 cagagcttca tcgagcggat gaccggcttc gataagaacc tgcccaacga gaaggtgctg
     5041 cccaagcaca gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa
     5101 tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc
     5161 gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac
     5221 ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac
     5281 gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac
     5341 aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac
     5401 agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg
     5461 aagcagctga agcggcggag atacaccggc tggggcgccc tgagccggaa gctgatcaac
     5521 ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc
     5581 gccaacagaa acttcatggc cctgatccac gacgacagcc tgacctttaa agaggacatc
     5641 cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc
     5701 ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg
     5761 aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag
     5821 accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc
     5881 aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac
     5941 gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg
     6001 gacatcaacc ggctgtccga ctacgatgtg gaccatatcg tgcctcagag ctttctgaag
     6061 gacgactcca tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg caagagcgac
     6121 aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcggca gctgctgaac
     6181 gccaagctga ttacccagag aaagttcgac aatctgacca aggccgagag aggcggcctg
     6241 agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg ggccatcaca
     6301 aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga gaatgacaag
     6361 ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag
     6421 gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac
     6481 ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc
     6541 gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcaggaa
     6601 atcggcaagg ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc
     6661 gagattaccc tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa
     6721 accggggaga tcgtgtggga taagggccgg gattttgcca ccgtgcggaa agtgctgagc
     6781 atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag
     6841 tctatcctgc ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct
     6901 aagaagtacg gcggcttcga cagccccacc gtggcctatt ctgtgctggt ggtggccaaa
     6961 gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc
     7021 atggaaagaa gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa
     7081 gaagtgaaaa aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac
     7141 ggccggaaga gaatgctggc ctctgccggc gaactgcaga agggaaacga actggccctg
     7201 ccctccaaat atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc
     7261 cccgaggata atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag
     7321 atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac
     7381 aaagtgctgt ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat
     7441 atcatccacc tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac
     7501 accaccatcg accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc
     7561 caccagagca tcaccggcct gtacgagaca cgaatcgatt tgtctcagct gggaggcgac
     7621 gccgcaaaaa ggccggcggc cacgaaaaag gccggccagg caaaaaagaa aaaggcaggc
     7681 tcaggcgagg gcagaggaag tctgctaaca tgcggtgacg tcgaggagaa tcctggccca
     7741 agcgctatgg ccaagccttt gtctcaagaa gaatccaccc tcattgaaag agcaacggct
     7801 acaatcaaca gcatccccat ctctgaagac tacagcgtcg ccagcgcagc tctctctagc
     7861 gacggccgca tcttcactgg tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa
     7921 ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca acctgacttg tatcgtcgcg
     7981 atcggaaatg agaacagggg catcttgagc ccctgcggac ggtgccgaca ggtgcttctc
     8041 gatctgcatc ctgggatcaa agccatagtg aaggacagtg atggacagcc gacggcagtt
     8101 gggattcgtg aattgctgcc ctctggttat gtgtgggagg gcgcaacaaa cttctctctg
     8161 ctgaaacaag ccggagatgt cgaagagaat cctggaccga ctagtatggt gagcgagctg
     8221 attaaggaga acatgcacat gaagctatac atggagggca ccgtgaacaa ccaccacttc
     8281 aagtgcacat ccgagggcga aggcaagccc tacgagggca cccagaccat gagaatcaag
     8341 gcggtcgagg gcggccctct ccccttcgcc ttcgacatcc tggctaccag cttcatgtac
     8401 ggcagcaaaa ccttcatcaa ccacacccag ggcatccccg acttctttaa gcagtccttc
     8461 cccgagggct tcacatggga gagagtcacc acatacgaag acgggggcgt gctgaccgct
     8521 acccaggaca ccagcctcca ggacggctgc ctcatctaca acgtcaagat cagaggggtg
     8581 aacttcccat ccaacggccc tgtgatgcag aagaaaacac tcggctggga ggcctccacc
     8641 gagaccctgt accccgctga cggcggcctg gaaggcagag ccgacatggc cctgaagctc
     8701 gtgggcgggg gccacctgat ctgcaacttg aagaccacat acagatccaa gaaacccgct
     8761 aagaacctca agatgcccgg cgtctactat gtggacagaa gactggaaag aatcaaggag
     8821 gccgacaaag agacctacgt cgagcagcac gaggtggctg tggccagata ctgcgacctc
     8881 cctagcaaac tggggcacag atgatagacg cgttaagtcg acaatcaacc tctggattac
     8941 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     9001 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     9061 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     9121 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     9181 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     9241 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     9301 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     9361 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     9421 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     9481 agtcggatct ccctttgggc cgcctccccg cgtcgacttt aagaccaatg acttacaagg
     9541 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     9601 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     9661 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     9721 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     9781 tcagaccctt ttagtcagtg tggaaaatct ctagcagtac gtatagtagt tcatgtcatc
     9841 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt
     9901 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     9961 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
    10021 tctggctcta gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
    10081 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
    10141 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta
    10201 cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt
    10261 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
    10321 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
    10381 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
    10441 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
    10501 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
    10561 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
    10621 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
    10681 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
    10741 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
    10801 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
    10861 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
    10921 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
    10981 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
    11041 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
    11101 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
    11161 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
    11221 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
    11281 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
    11341 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
    11401 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
    11461 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
    11521 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
    11581 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
    11641 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
    11701 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
    11761 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
    11821 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
    11881 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
    11941 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
    12001 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
    12061 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
    12121 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
    12181 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
    12241 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
    12301 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
    12361 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
    12421 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
    12481 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
    12541 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
    12601 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
    12661 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
    12721 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
    12781 aagagcgccc aatacgc
//
