LOCUS       sequence-342246-       13011 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing Cas9 and a control sgRNA that targets a
            safe harbor site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13011)
  AUTHORS   Chaikovsky AC, Li C, Jeng EE, Loebell S, Lee MC, Murray CW, Cheng R,
            Demeter J, Swaney DL, Chen SH, Newton BW, Johnson JR, Drainas AP, 
            Shue YT, Seoane JA, Srinivasan P, He A, Yoshida A, Hipkins SQ, 
            McCrea E, Poltorack CD, Krogan NJ, Diehl JA, Kong C, Jackson PK, 
            Curtis C, Petrov DA, Bassik MC, Winslow MM, Sage J
  TITLE     The AMBRA1 E3 ligase adaptor regulates the stability of cyclin D.
  JOURNAL   Nature. 2021 Apr;592(7856):794-798. doi: 10.1038/s41586-021-03474-7.
            Epub 2021 Apr 14.
  PUBMED    33854239
REFERENCE   2  (bases 1 to 13011)
  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"; doi: 
            "10.1038/s41586-021-03474-7"; journalName: "Nature"; date: 
            "2021-04"; volume: "592"; issue: "7856"; pages: "794-798"
COMMENT     Sequence Label: lentiCRISPRv2_sgCtrl#1
FEATURES             Location/Qualifiers
     source          1..13011
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2608..2856
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2608..2628
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2779..2798
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2876..2951
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3013..3224
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3243..3252
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3249..7352
                     /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             3249..7352
                     /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             7353..7400
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7401..7424
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             7434..7490
                     /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             7491..8087
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     7984..8004
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8103..8691
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8156..8176)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8574..8585)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8763..8996
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9022..9039)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9028..9252
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9298..9726
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9385..9404)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9595..9616
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9735..9755)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9740..10069
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9920..10055
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9982..10001
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10117..10164
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10183..10557
                     /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    10687..10808
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10724..10743)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10778..10797
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10857..10873)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10857..10873)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10870..10892)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10881..10897
                     /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(10905..10911,10912..10929,10930..10935))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10950..10971
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11088..11105)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11259..11847)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11339..11358)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12018..12809,12810..12878))
                     /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     12641..12660
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12879..12983)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta aggtaccgag ggcctatttc ccatgattcc ttcatatttg
     2641 catatacgat acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag
     2701 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta
     2761 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc
     2821 ttggctttat atatcttgtg gaaaggacga aacaccgtga gcctatatta attgggtttt
     2881 agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac
     2941 cgagtcggtg cttttttgaa ttcgctagct aggtcttgaa aggagtggga attggctccg
     3001 gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg
     3061 tcggcaattg atccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg
     3121 tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg
     3181 ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggaccgg ttctagagcg
     3241 ctgccaccat ggacaagaag tacagcatcg gcctggacat cggcaccaac tctgtgggct
     3301 gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca
     3361 ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac agcggcgaaa
     3421 cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc
     3481 ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct
     3541 tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca
     3601 tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc
     3661 tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc
     3721 tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca
     3781 acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg
     3841 aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca
     3901 agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt
     3961 tcggaaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc
     4021 tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc
     4081 tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg
     4141 acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga
     4201 gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc
     4261 tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg
     4321 gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag ttcatcaagc
     4381 ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc
     4441 tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag
     4501 agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg
     4561 aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg
     4621 gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact
     4681 tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg atgaccaact
     4741 tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact
     4801 tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg
     4861 ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag accaaccgga
     4921 aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg
     4981 tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc
     5041 tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag
     5101 atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa
     5161 cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg
     5221 gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga
     5281 caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc
     5341 acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg
     5401 atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc
     5461 tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga
     5521 acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag aagaacagcc
     5581 gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag
     5641 aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga
     5701 atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg
     5761 tggaccatat cgtgcctcag agctttctga aggacgactc catcgacaac aaggtgctga
     5821 ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga
     5881 agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag agaaagttcg
     5941 acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca
     6001 agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc
     6061 ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc
     6121 tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga
     6181 tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga
     6241 tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg
     6301 tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc aagtacttct
     6361 tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac ggcgagatcc
     6421 ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg gataagggcc
     6481 gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga
     6541 ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg aacagcgata
     6601 agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc gacagcccca
     6661 ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc aagaaactga
     6721 agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc gagaagaatc
     6781 ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc
     6841 tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctctgccg
     6901 gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc
     6961 tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt
     7021 ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc gagttctcca
     7081 agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac aacaagcacc
     7141 gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc
     7201 tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag aggtacacca
     7261 gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga
     7321 cacggatcga cctgtctcag ctgggaggcg acaagcgacc tgccgccaca aagaaggctg
     7381 gacaggctaa gaagaagaaa gattacaaag acgatgacga taagggatcc ggcgcaacaa
     7441 acttctctct gctgaaacaa gccggagatg tcgaagagaa tcctggaccg accgagtaca
     7501 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
     7561 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
     7621 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     7681 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     7741 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
     7801 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     7861 tggccaccgt cggagtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     7921 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
     7981 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     8041 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaacg cgttaagtcg
     8101 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     8161 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     8221 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     8281 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     8341 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     8401 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     8461 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     8521 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     8581 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     8641 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cgtcgacttt
     8701 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg
     8761 actggaaggg ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc
     8821 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     8881 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     8941 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagggc
     9001 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     9061 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     9121 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     9181 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     9241 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     9301 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     9361 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     9421 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     9481 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     9541 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     9601 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     9661 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     9721 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     9781 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     9841 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     9901 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     9961 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
    10021 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
    10081 agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag
    10141 tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt
    10201 gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg
    10261 ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg
    10321 accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg
    10381 tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc
    10441 cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc
    10501 gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac
    10561 gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
    10621 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
    10681 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
    10741 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
    10801 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
    10861 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
    10921 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
    10981 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
    11041 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
    11101 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
    11161 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
    11221 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
    11281 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
    11341 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
    11401 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
    11461 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
    11521 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
    11581 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
    11641 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
    11701 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
    11761 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
    11821 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
    11881 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
    11941 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
    12001 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
    12061 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
    12121 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
    12181 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
    12241 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
    12301 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
    12361 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
    12421 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
    12481 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
    12541 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
    12601 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
    12661 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
    12721 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
    12781 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
    12841 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
    12901 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
    12961 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga c
//
