LOCUS       sequence-326558-       13489 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral construct for the expression of SpCas9 driven by EFS 
            promoter in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13489)
  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 13489)
  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_EFS-Cas9-P2A-HygR
FEATURES             Location/Qualifiers
     source          1..13489
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1443..1676
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1861..1905
                     /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    2203..2320
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2413..2624
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2643..2652
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2649..6752
                     /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             2649..6752
                     /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             6753..6800
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6801..6824
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6834..6890
                     /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             6891..7919
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MAKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     misc_feature    7944..8532
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7997..8017)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8415..8426)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8535..8551)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8536..8552)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             9057..9237
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9263..9280)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9269..9493
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9539..9967
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9626..9645)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9836..9857
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9976..9996)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9981..10310
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10161..10296
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10223..10242
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10358..10405
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10424..10798
                     /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    10928..11049
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10965..10984)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11019..11038
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11098..11114)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11098..11114)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11111..11133)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11122..11138
                     /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(11146..11152,11153..11170,11171..11176))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11191..11212
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11329..11346)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11500..12088)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11580..11599)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12259..13050,13051..13119))
                     /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     12882..12901
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13120..13224)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(13299..13318)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2341 cagagatcca gtttggttaa ttagctagct aggtcttgaa aggagtggga attggctccg
     2401 gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg
     2461 tcggcaattg atccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg
     2521 tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg
     2581 ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggaccgg ttctagagcg
     2641 ctgccaccat ggacaagaag tacagcatcg gcctggacat cggcaccaac tctgtgggct
     2701 gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca
     2761 ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac agcggcgaaa
     2821 cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc
     2881 ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct
     2941 tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca
     3001 tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc
     3061 tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc
     3121 tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca
     3181 acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg
     3241 aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca
     3301 agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt
     3361 tcggaaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc
     3421 tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc
     3481 tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg
     3541 acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga
     3601 gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc
     3661 tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg
     3721 gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag ttcatcaagc
     3781 ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc
     3841 tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag
     3901 agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg
     3961 aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg
     4021 gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact
     4081 tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg atgaccaact
     4141 tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact
     4201 tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg
     4261 ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag accaaccgga
     4321 aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg
     4381 tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc
     4441 tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag
     4501 atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa
     4561 cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg
     4621 gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga
     4681 caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc
     4741 acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg
     4801 atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc
     4861 tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga
     4921 acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag aagaacagcc
     4981 gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag
     5041 aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga
     5101 atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg
     5161 tggaccatat cgtgcctcag agctttctga aggacgactc catcgacaac aaggtgctga
     5221 ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga
     5281 agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag agaaagttcg
     5341 acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca
     5401 agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc
     5461 ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc
     5521 tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga
     5581 tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga
     5641 tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg
     5701 tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc aagtacttct
     5761 tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac ggcgagatcc
     5821 ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg gataagggcc
     5881 gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga
     5941 ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg aacagcgata
     6001 agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc gacagcccca
     6061 ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc aagaaactga
     6121 agagtgtgaa agagctactg gggatcacca tcatggaaag aagcagcttc gagaagaatc
     6181 ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc
     6241 tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctctgccg
     6301 gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc
     6361 tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt
     6421 ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc gagttctcca
     6481 agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac aacaagcacc
     6541 gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc
     6601 tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag aggtacacca
     6661 gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga
     6721 cacggatcga cctgtctcag ctgggaggcg acaagcgacc tgccgccaca aagaaggctg
     6781 gacaggctaa gaagaagaaa gattacaaag acgatgacga taagggatcc ggcgcaacaa
     6841 acttctctct gctgaaacaa gccggagatg tcgaagagaa tcctggaccg atggccaaaa
     6901 agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtgt
     6961 ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag
     7021 ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg
     7081 tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac attggggagt
     7141 tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc
     7201 tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggcaatg gatgcgatcg
     7261 ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc
     7321 aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc
     7381 aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc
     7441 tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca
     7501 atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg
     7561 gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg
     7621 agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg ccgcggctcc
     7681 gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt
     7741 tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga
     7801 ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag
     7861 aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaatgag
     7921 aattcgatat caagcttatc ggtaatcaac ctctggatta caaaatttgt gaaagattga
     7981 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     8041 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     8101 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     8161 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     8221 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     8281 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     8341 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     8401 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     8461 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     8521 ccgcctcccc gcatcgatac cgtcgacctc gagacctaga aaaacatgga gcaatcacaa
     8581 gtagcaatac agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg
     8641 aggtgggttt tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg
     8701 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     8761 gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc cctgattggc
     8821 agaactacac accagggcca gggatcagat atccactgac ctttggatgg tgctacaagc
     8881 tagtaccagt tgagcaagag aaggtagaag aagccaatga aggagagaac acccgcttgt
     8941 tacaccctgt gagcctgcat gggatggatg acccggagag agaagtatta gagtggaggt
     9001 ttgacagccg cctagcattt catcacatgg cccgagagct gcatccggac tgtactgggt
     9061 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     9121 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     9181 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcaggg
     9241 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     9301 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     9361 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     9421 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     9481 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     9541 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     9601 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     9661 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     9721 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     9781 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     9841 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     9901 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     9961 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
    10021 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
    10081 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
    10141 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
    10201 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
    10261 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
    10321 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata
    10381 gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag
    10441 tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg
    10501 gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt
    10561 gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt
    10621 gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt
    10681 ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt
    10741 cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca
    10801 cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
    10861 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
    10921 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
    10981 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
    11041 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
    11101 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
    11161 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
    11221 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
    11281 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
    11341 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
    11401 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
    11461 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
    11521 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
    11581 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
    11641 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
    11701 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
    11761 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
    11821 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
    11881 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
    11941 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
    12001 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
    12061 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
    12121 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
    12181 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
    12241 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
    12301 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
    12361 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
    12421 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
    12481 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
    12541 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
    12601 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
    12661 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
    12721 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
    12781 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
    12841 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
    12901 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
    12961 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
    13021 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
    13081 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
    13141 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
    13201 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
    13261 atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca
    13321 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
    13381 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
    13441 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgtt
//
