LOCUS       sequence-323780-       13724 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral construct for the expression of SpCas9 driven by PGK 
            promoter in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13724)
  AUTHORS   Park J, Lim JM, Jung I, Heo SJ, Park J, Chang Y, Kim HK, Jung D, Yu 
            JH, Min S, Yoon S, Cho SR, Park T, Kim HH
  TITLE     Recording of elapsed time and temporal information about biological 
            events using Cas9.
  JOURNAL   Cell. 2021 Jan 28. pii: S0092-8674(21)00014-3. doi: 
            10.1016/j.cell.2021.01.014.
  PUBMED    33539780
REFERENCE   2  (bases 1 to 13724)
  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: "Cell. 2021 
            Jan 28. pii: S0092-8674(21)00014-3. doi: 
            10.1016/j.cell.2021.01.014."
COMMENT     Sequence Label: Lenti_PGK-Cas9-P2A-HygR
FEATURES             Location/Qualifiers
     source          1..13724
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..200
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             218..398
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    445..570
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1063..1296
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1481..1525
                     /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    1823..1940
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1990..2489
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2025..2048)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2422..2441
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      2498..2507
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2504..6607
                     /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             2504..6607
                     /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             6608..6655
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6656..6679
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6689..6745
                     /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             6746..7774
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MAKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     misc_feature    7799..8387
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7852..7872)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8270..8281)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8390..8406)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8391..8407)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             8912..9092
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9118..9135)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9124..9348
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9394..9822
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9481..9500)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9691..9712
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9831..9851)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9836..10165
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10016..10151
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10078..10097
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10213..10260
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10279..10653
                     /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    10783..10904
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10820..10839)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10874..10893
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10953..10969)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10953..10969)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10966..10988)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10977..10993
                     /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(11001..11007,11008..11025,11026..11031))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11046..11067
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11184..11201)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11355..11943)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11435..11454)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12114..12905,12906..12974))
                     /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     12737..12756
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12975..13079)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(13154..13173)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        13345..13724
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag cgcgttttgc ctgtactggg tctctctggt tagaccagat
      241 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
      301 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
      361 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg
      421 aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc
      481 acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc
      541 tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga
      601 tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta
      661 tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa
      721 ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt
      781 agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa
      841 gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca
      901 cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
      961 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1021 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1081 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1141 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1201 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1261 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1321 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1381 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1441 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1501 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1561 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1621 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1681 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1741 cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac
     1801 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
     1861 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
     1921 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttaa
     1981 ttagctagcg ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag
     2041 ccccgctggg cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca
     2101 ccggtaggcg ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc
     2161 ccctagtcag gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg
     2221 aagtagcacg tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg
     2281 taggcctttg gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc
     2341 tgggaagggg tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc
     2401 gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt
     2461 tctcctcttc ctcatctccg ggcctttcga tagcgctgcc accatggaca agaagtacag
     2521 catcggcctg gacatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa
     2581 ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa
     2641 cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag
     2701 aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt
     2761 cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag agtccttcct
     2821 ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt
     2881 ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac
     2941 cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg
     3001 ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat
     3061 ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt
     3121 ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat
     3181 cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg ccctgagcct
     3241 gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct
     3301 gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta
     3361 cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct
     3421 gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga
     3481 cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa
     3541 gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg
     3601 agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac
     3661 cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga
     3721 caacggcagc atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca
     3781 ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt
     3841 ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac
     3901 cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc
     3961 ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa
     4021 ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa
     4081 agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa
     4141 ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga
     4201 ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg
     4261 gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt
     4321 cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt
     4381 tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa
     4441 agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct
     4501 gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga
     4561 cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga
     4621 ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa
     4681 tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga
     4741 gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga
     4801 gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga
     4861 gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct
     4921 gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca
     4981 ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc ctcagagctt
     5041 tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga accggggcaa
     5101 gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct
     5161 gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg
     5221 cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca
     5281 gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa
     5341 tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt
     5401 ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga
     5461 cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag
     5521 cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga
     5581 gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt
     5641 caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa
     5701 cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt
     5761 gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag
     5821 caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa agaaggactg
     5881 ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg tgctggtggt
     5941 ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc tactggggat
     6001 caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg
     6061 ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct
     6121 ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact
     6181 ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa
     6241 gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct
     6301 ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa
     6361 tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc
     6421 cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta
     6481 ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc tggacgccac
     6541 cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg
     6601 aggcgacaag cgacctgccg ccacaaagaa ggctggacag gctaagaaga agaaagatta
     6661 caaagacgat gacgataagg gatccggcgc aacaaacttc tctctgctga aacaagccgg
     6721 agatgtcgaa gagaatcctg gaccgatggc caaaaagcct gaactcaccg cgacgtctgt
     6781 cgagaagttt ctgatcgaaa agttcgacag cgtgtccgac ctgatgcagc tctcggaggg
     6841 cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa
     6901 tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg catcggccgc
     6961 gctcccgatt ccggaagtgc ttgacattgg ggagttcagc gagagcctga cctattgcat
     7021 ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt
     7081 tctgcagccg gtcgcggagg caatggatgc gatcgctgcg gccgatctta gccagacgag
     7141 cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat
     7201 atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag
     7261 tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt
     7321 ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat
     7381 aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa
     7441 catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg
     7501 gaggcatccg gagcttgcag gatcgccgcg gctccgggcg tatatgctcc gcattggtct
     7561 tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg
     7621 tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg
     7681 cagaagcgcg gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg
     7741 acgccccagc actcgtccga gggcaaagga atgagaattc gatatcaagc ttatcggtaa
     7801 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     7861 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     7921 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     7981 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     8041 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     8101 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     8161 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     8221 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     8281 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     8341 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg
     8401 acctcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc
     8461 tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca
     8521 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     8581 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg
     8641 gatctaccac acacaaggct acttccctga ttggcagaac tacacaccag ggccagggat
     8701 cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc aagagaaggt
     8761 agaagaagcc aatgaaggag agaacacccg cttgttacac cctgtgagcc tgcatgggat
     8821 ggatgacccg gagagagaag tattagagtg gaggtttgac agccgcctag catttcatca
     8881 catggcccga gagctgcatc cggactgtac tgggtctctc tggttagacc agatctgagc
     8941 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     9001 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     9061 acccttttag tcagtgtgga aaatctctag cagggcccgt ttaaacccgc tgatcagcct
     9121 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     9181 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     9241 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     9301 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     9361 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     9421 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     9481 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     9541 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     9601 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     9661 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     9721 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     9781 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     9841 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     9901 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     9961 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
    10021 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
    10081 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
    10141 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
    10201 tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac
    10261 aaggtgagga actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc
    10321 gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg
    10381 gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag
    10441 gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg
    10501 tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg
    10561 accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac
    10621 tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc
    10681 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
    10741 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
    10801 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
    10861 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
    10921 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
    10981 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
    11041 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
    11101 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
    11161 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
    11221 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
    11281 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
    11341 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
    11401 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
    11461 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
    11521 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
    11581 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
    11641 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
    11701 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
    11761 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
    11821 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
    11881 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
    11941 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
    12001 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
    12061 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
    12121 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
    12181 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
    12241 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
    12301 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
    12361 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
    12421 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
    12481 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
    12541 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
    12601 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
    12661 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
    12721 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
    12781 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
    12841 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
    12901 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
    12961 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
    13021 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
    13081 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
    13141 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
    13201 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
    13261 gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg
    13321 atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa
    13381 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
    13441 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
    13501 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
    13561 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
    13621 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
    13681 ctacttggca gtacatctac gtattagtca tcgctattac catg
//
