LOCUS       sequence-155356-       14118 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral construct for building stable cell lines expressing Cas9 
            via puromycin selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14118)
  AUTHORS   Perli SD, Cui CH, Lu TK
  TITLE     Continuous genetic recording with self-targeting CRISPR-Cas in human
            cells.
  JOURNAL   Science. 2016 Aug 18. pii: aag0511.
  PUBMED    27540006
REFERENCE   2  (bases 1 to 14118)
  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: "Science. 
            2016 Aug 18. pii: aag0511."
COMMENT     Sequence Label: Construct 12 - hUBCp_Cas9_3xNLS_p2a_puroR
FEATURES             Location/Qualifiers
     source          1..14118
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             174..218
                     /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    516..633
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        687..1899
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1736..1755
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      1957..1966
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1963..6066
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             1963..6066
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             6067..6114
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6139..6159
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6166..6186
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6205..6261
                     /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             6262..6858
                     /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     6755..6775
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6886..7474
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6939..6959)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7357..7368)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7477..7493)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7478..7494)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             7999..8179
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8205..8222)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8211..8435
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8481..8909
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8568..8587)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8778..8799
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8918..8938)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8923..9252
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9103..9238
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9165..9184
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9300..9347
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9366..9740
                     /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    9870..9991
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9907..9926)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9961..9980
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10040..10056)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10040..10056)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10053..10075)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10064..10080
                     /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(10088..10094,10095..10112,10113..10118))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10133..10154
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10271..10288)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10442..11030)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10522..10541)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11201..11992,11993..12061))
                     /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     11824..11843
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12062..12166)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12241..12260)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        12432..12811
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        12813..13011
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     12966..12986
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             13029..13209
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    13256..13381
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    13874..14107
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
       61 aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca
      121 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg
      181 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa
      241 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa
      301 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt
      361 ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag
      421 gtggagagag agacagagac agatccattc gattagtgaa cggatcggca ctgcgtgcgc
      481 caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg
      541 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
      601 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat
      661 ccagtttggt taattaaggg tgcagcggcc tccgcgccgg gttttggcgc ctcccgcggg
      721 cgcccccctc ctcacggcga gcgctgccac gtcagacgaa gggcgcagcg agcgtcctga
      781 tccttccgcc cggacgctca ggacagcggc ccgctgctca taagactcgg ccttagaacc
      841 ccagtatcag cagaaggaca ttttaggacg ggacttgggt gactctaggg cactggtttt
      901 ctttccagag agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg
      961 atctccgtgg ggcggtgaac gccgatgatt atataaggac gcgccgggtg tggcacagct
     1021 agttccgtcg cagccgggat ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca
     1081 cttggtgagt agcgggctgc tgggctggcc ggggctttcg tggccgccgg gccgctcggt
     1141 gggacggaag cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg
     1201 ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg
     1261 gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg
     1321 gcaagaaccc aaggtcttga ggccttcgct aatgcgggaa agctcttatt cgggtgagat
     1381 gggctggggc accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggg
     1441 tttgtcgtct gttgcggggg cggcagttat ggcggtgccg ttgggcagtg cacccgtacc
     1501 tttgggagcg cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca
     1561 gggtggggcc acctgccggt aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt
     1621 tcgggcctag ggtaggctct cctgaatcga caggcgccgg acctctggtg aggggaggga
     1681 taagtgaggc gtcagtttct ttggtcggtt ttatgtacct atcttcttaa gtagctgaag
     1741 ctccggtttt gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca
     1801 ccttttgaaa tgtaatcatt tgggtcaata tgtaattttc agtgttagac tagtaaattg
     1861 tccgctaaat tctggccgtt tttggctttt ttgttagacg aagcttgggc tgcaggtcga
     1921 ctctagagga tccccgggta ccggtcgcca acgcgtgcca ccatggacaa gaagtacagc
     1981 atcggcctgg acatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag
     2041 gtgcccagca agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac
     2101 ctgatcggag ccctgctgtt cgacagcggc gaaacagccg aggccacccg gctgaagaga
     2161 accgccagaa gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc
     2221 agcaacgaga tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg
     2281 gtggaagagg ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg
     2341 gcctaccacg agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc
     2401 gacaaggccg acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc
     2461 cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc
     2521 cagctggtgc agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg
     2581 gacgccaagg ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc
     2641 gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa acctgattgc cctgagcctg
     2701 ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg
     2761 agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac
     2821 gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg
     2881 agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac
     2941 gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag
     3001 tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat tgacggcgga
     3061 gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc
     3121 gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg gaccttcgac
     3181 aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag
     3241 gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc
     3301 cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc
     3361 agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgct
     3421 tccgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag
     3481 gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtataacga gctgaccaaa
     3541 gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaag
     3601 gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag
     3661 gactacttca agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg
     3721 ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc
     3781 ctggacaatg aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt
     3841 gaggacagag agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa
     3901 gtgatgaagc agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg
     3961 atcaacggca tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac
     4021 ggcttcgcca acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag
     4081 gacatccaga aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat
     4141 ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag
     4201 ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag
     4261 aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag
     4321 ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg
     4381 cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag
     4441 gaactggaca tcaaccggct gtccgactac gatgtggacc atatcgtgcc tcagagcttt
     4501 ctgaaggacg actccatcga caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag
     4561 agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg gcggcagctg
     4621 ctgaacgcca agctgattac ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc
     4681 ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag
     4741 atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaat
     4801 gacaagctga tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc
     4861 cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac
     4921 gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc
     4981 gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag
     5041 caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc
     5101 aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac
     5161 ggcgaaaccg gggagatcgt gtgggataag ggccgggatt ttgccaccgt gcggaaagtg
     5221 ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc
     5281 aaagagtcta tcctgcccaa gaggaacagc gataagctga tcgccagaaa gaaggactgg
     5341 gaccctaaga agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg
     5401 gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc
     5461 accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc
     5521 tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg
     5581 gaaaacggcc ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg
     5641 gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag
     5701 ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa gcactacctg
     5761 gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat
     5821 ctggacaaag tgctgtccgc ctacaacaag caccgggata agcccatcag agagcaggcc
     5881 gagaatatca tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac
     5941 tttgacacca ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc
     6001 ctgatccacc agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga
     6061 ggcgacaagc gtcctgctgc tactaagaaa gctggtcaag ctaagaaaaa gaaagctagc
     6121 ggcagcggcg ccggatcccc aaagaagaaa aggaaggttg aagaccccaa gaaaaagagg
     6181 aaggtgataa gcgctggaag cggagctact aacttcagcc tgctgaagca ggctggagac
     6241 gtggaggaga accctggacc taccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     6301 gacgtccccc gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     6361 cacaccgtcg acccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     6421 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     6481 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     6541 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg
     6601 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
     6661 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc
     6721 ggggtgcccg ccttcctgga gacatccgcg ccccgcaacc tccccttcta cgagcggctc
     6781 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     6841 cgcaagcccg gtgcctgata agaattcgat atcaagctta tcgataatca acctctggat
     6901 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     6961 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     7021 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     7081 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     7141 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     7201 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     7261 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
     7321 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     7381 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     7441 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc tcgagaccta
     7501 gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg
     7561 ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga
     7621 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg
     7681 gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat ctaccacaca
     7741 caaggctact tccctgattg gcagaactac acaccagggc cagggatcag atatccactg
     7801 acctttggat ggtgctacaa gctagtacca gttgagcaag agaaggtaga agaagccaat
     7861 gaaggagaga acacccgctt gttacaccct gtgagcctgc atgggatgga tgacccggag
     7921 agagaagtat tagagtggag gtttgacagc cgcctagcat ttcatcacat ggcccgagag
     7981 ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     8041 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     8101 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     8161 gtgtggaaaa tctctagcag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     8221 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     8281 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     8341 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     8401 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     8461 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     8521 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     8581 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     8641 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     8701 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     8761 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     8821 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     8881 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     8941 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     9001 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     9061 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     9121 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     9181 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     9241 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg
     9301 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact
     9361 aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag
     9421 cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg
     9481 ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc
     9541 cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt
     9601 cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg
     9661 agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg
     9721 tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg
     9781 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     9841 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     9901 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     9961 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
    10021 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
    10081 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
    10141 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
    10201 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
    10261 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
    10321 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
    10381 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
    10441 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
    10501 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
    10561 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
    10621 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
    10681 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
    10741 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
    10801 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
    10861 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
    10921 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
    10981 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
    11041 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
    11101 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
    11161 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
    11221 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
    11281 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
    11341 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
    11401 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
    11461 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
    11521 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
    11581 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
    11641 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
    11701 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
    11761 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
    11821 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
    11881 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
    11941 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
    12001 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
    12061 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
    12121 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
    12181 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
    12241 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
    12301 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
    12361 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
    12421 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
    12481 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
    12541 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
    12601 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
    12661 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
    12721 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
    12781 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
    12841 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
    12901 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
    12961 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg
    13021 cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
    13081 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
    13141 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
    13201 tctctagcag tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc
    13261 tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt
    13321 gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt
    13381 cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg
    13441 aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc
    13501 agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca
    13561 accatccctt cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct
    13621 ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag acaagataga
    13681 ggaagagcaa aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg
    13741 gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa
    13801 ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa
    13861 gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg
    13921 gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc
    13981 agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct
    14041 ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac
    14101 agctcctggg gatttggg
//
