LOCUS       sequence-323777-       13882 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral construct for the expression of SpCas9 driven by CMV 
            promoter in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13882)
  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 13882)
  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_CMV-Cas9-P2A-HygR
FEATURES             Location/Qualifiers
     source          1..13882
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        28..226
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             244..424
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    471..596
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1089..1322
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1507..1551
                     /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    1849..1966
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2020..2399
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2400..2603
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2648..2667
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2648..2666
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2682..2691
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2688..6791
                     /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             2688..6791
                     /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             6792..6839
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6840..6863
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6873..6929
                     /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             6930..7958
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MAKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     misc_feature    7983..8571
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8036..8056)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8454..8465)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8574..8590)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8575..8591)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             9096..9276
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9302..9319)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9308..9532
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9578..10006
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9665..9684)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9875..9896
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10015..10035)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10020..10349
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10200..10335
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10262..10281
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10397..10444
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10463..10837
                     /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    10967..11088
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11004..11023)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11058..11077
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11137..11153)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11137..11153)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11150..11172)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11161..11177
                     /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(11185..11191,11192..11209,11210..11215))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11230..11251
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11368..11385)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11539..12127)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11619..11638)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12298..13089,13090..13158))
                     /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     12921..12940
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13159..13263)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(13338..13357)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(13529..13882,1..26)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
       61 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      121 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      181 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt
      241 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
      301 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
      361 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
      421 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
      481 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
      541 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
      601 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
      661 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
      721 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
      781 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
      841 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
      901 agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga
      961 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
     1021 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
     1081 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
     1141 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
     1201 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
     1261 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
     1321 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
     1381 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
     1441 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
     1501 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
     1561 tttaacataa caaattggct gtggtatata aaattattca taatgatagt aggaggcttg
     1621 gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat
     1681 tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga
     1741 atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatcg
     1801 gcactgcgtg cgccaattct gcagacaaat ggcagtattc atccacaatt ttaaaagaaa
     1861 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
     1921 acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag
     1981 ggacagcaga gatccagttt ggttaattag ctacgcgttg acattgatta ttgactagtt
     2041 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     2101 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     2161 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     2221 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     2281 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     2341 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     2401 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     2461 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     2521 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     2581 gggaggtcta tataagcaga gctctctggc taactagaga acccactgct tactggctta
     2641 tcgaaattaa tacgactcac tatagggaga cccaagctgc tgccaccatg gacaagaagt
     2701 acagcatcgg cctggacatc ggcaccaact ctgtgggctg ggccgtgatc accgacgagt
     2761 acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga
     2821 agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc acccggctga
     2881 agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat ctgcaagaga
     2941 tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg gaagagtcct
     3001 tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac atcgtggacg
     3061 aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa ctggtggaca
     3121 gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg atcaagttcc
     3181 ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg gacaagctgt
     3241 tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc aacgccagcg
     3301 gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc
     3361 tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg attgccctga
     3421 gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat gccaaactgc
     3481 agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag atcggcgacc
     3541 agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg ctgagcgaca
     3601 tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg atcaagagat
     3661 acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg
     3721 agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc tacattgacg
     3781 gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa aagatggacg
     3841 gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag cagcggacct
     3901 tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc attctgcggc
     3961 ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag aagatcctga
     4021 ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga ttcgcctgga
     4081 tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg gtggacaagg
     4141 gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac ctgcccaacg
     4201 agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat aacgagctga
     4261 ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc ggcgagcaga
     4321 aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga
     4381 aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag
     4441 atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc aaggacaagg
     4501 acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg accctgacac
     4561 tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg
     4621 acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg ctgagccgga
     4681 agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat ttcctgaagt
     4741 ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc ctgaccttta
     4801 aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac gagcacattg
     4861 ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg aaggtggtgg
     4921 acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc gaaatggcca
     4981 gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg aagcggatcg
     5041 aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc
     5101 agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat atgtacgtgg
     5161 accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc gtgcctcaga
     5221 gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac aagaaccggg
     5281 gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac tactggcggc
     5341 agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc aaggccgaga
     5401 gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg gtggaaaccc
     5461 ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact aagtacgacg
     5521 agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg
     5581 atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac caccacgccc
     5641 acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg
     5701 aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg atcgccaaga
     5761 gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac atcatgaact
     5821 ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct ctgatcgaga
     5881 caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc accgtgcgga
     5941 aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct
     6001 tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc agaaagaagg
     6061 actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat tctgtgctgg
     6121 tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa gagctactgg
     6181 ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt ctggaagcca
     6241 agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac tccctgttcg
     6301 agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag aagggaaacg
     6361 aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac tatgagaagc
     6421 tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag cacaagcact
     6481 acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc ctggccgacg
     6541 ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc atcagagagc
     6601 aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct gccgccttca
     6661 agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag gtgctggacg
     6721 ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac ctgtctcagc
     6781 tgggaggcga caagcgacct gccgccacaa agaaggctgg acaggctaag aagaagaaag
     6841 attacaaaga cgatgacgat aagggatccg gcgcaacaaa cttctctctg ctgaaacaag
     6901 ccggagatgt cgaagagaat cctggaccga tggccaaaaa gcctgaactc accgcgacgt
     6961 ctgtcgagaa gtttctgatc gaaaagttcg acagcgtgtc cgacctgatg cagctctcgg
     7021 agggcgaaga atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg
     7081 taaatagctg cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatcgg
     7141 ccgcgctccc gattccggaa gtgcttgaca ttggggagtt cagcgagagc ctgacctatt
     7201 gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg
     7261 ctgttctgca gccggtcgcg gaggcaatgg atgcgatcgc tgcggccgat cttagccaga
     7321 cgagcgggtt cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt
     7381 tcatatgcgc gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg
     7441 tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg
     7501 aagtccggca cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc
     7561 gcataacagc ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg
     7621 ccaacatctt cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg
     7681 agcggaggca tccggagctt gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg
     7741 gtcttgacca actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc
     7801 agggtcgatg cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg
     7861 cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa
     7921 accgacgccc cagcactcgt ccgagggcaa aggaatgaga attcgatatc aagcttatcg
     7981 gtaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     8041 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     8101 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     8161 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     8221 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     8281 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     8341 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     8401 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     8461 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     8521 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc
     8581 gtcgacctcg agacctagaa aaacatggag caatcacaag tagcaataca gcagctacca
     8641 atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt ccagtcacac
     8701 ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
     8761 aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat atccttgatc
     8821 tgtggatcta ccacacacaa ggctacttcc ctgattggca gaactacaca ccagggccag
     8881 ggatcagata tccactgacc tttggatggt gctacaagct agtaccagtt gagcaagaga
     8941 aggtagaaga agccaatgaa ggagagaaca cccgcttgtt acaccctgtg agcctgcatg
     9001 ggatggatga cccggagaga gaagtattag agtggaggtt tgacagccgc ctagcatttc
     9061 atcacatggc ccgagagctg catccggact gtactgggtc tctctggtta gaccagatct
     9121 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     9181 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     9241 tcagaccctt ttagtcagtg tggaaaatct ctagcagggc ccgtttaaac ccgctgatca
     9301 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     9361 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     9421 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     9481 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     9541 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     9601 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     9661 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     9721 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     9781 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     9841 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     9901 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     9961 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
    10021 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
    10081 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
    10141 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
    10201 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
    10261 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
    10321 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
    10381 gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata
    10441 cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc
    10501 gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt
    10561 cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt
    10621 ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga
    10681 gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc
    10741 catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg
    10801 caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag atttcgattc
    10861 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
    10921 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
    10981 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
    11041 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
    11101 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
    11161 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
    11221 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
    11281 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
    11341 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
    11401 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
    11461 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
    11521 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
    11581 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
    11641 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
    11701 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
    11761 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
    11821 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
    11881 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
    11941 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
    12001 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
    12061 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
    12121 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
    12181 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
    12241 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
    12301 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
    12361 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
    12421 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
    12481 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
    12541 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
    12601 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
    12661 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
    12721 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
    12781 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
    12841 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
    12901 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
    12961 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
    13021 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
    13081 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
    13141 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
    13201 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
    13261 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
    13321 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
    13381 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
    13441 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
    13501 cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa
    13561 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
    13621 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
    13681 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
    13741 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
    13801 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
    13861 tttcctactt ggcagtacat ct
//
