LOCUS       Exported               14975 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  SiC-V1 vector with SpCas9 gene and dTomato reporter. sgRNA targeting
            a gene of interest can be cloned downstream of U6 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    SiC-V1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14975)
  AUTHORS   Kelkar A, Zhu Y, Groth T, Stolfa G, Stablewski AB, Singhi N, Nemeth 
            M, Neelamegham S
  TITLE     Doxycycline-Dependent Self-Inactivation of CRISPR-Cas9 to Temporally
            Regulate On- and Off-Target Editing.
  JOURNAL   Mol Ther. 2019 Sep 12. pii: S1525-0016(19)30409-5. doi: 
            10.1016/j.ymthe.2019.09.006.
  PUBMED    31601489
REFERENCE   2  (bases 1 to 14975)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..14975
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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..2848
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     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        4738..4813
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4875..5086
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      5105..5114
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             5111..9214
                     /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             9215..9262
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9278..9334
                     /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             9341..10042
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(9474..9492)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     9742..9761
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     9936..9959
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    10067..10655
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10120..10140)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10538..10549)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10727..10960
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10986..11003)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10992..11216
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11262..11690
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11349..11368)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11559..11580
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11699..11719)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11704..12033
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11884..12019
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11946..11965
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12081..12128
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12147..12521
                     /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    12651..12772
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12688..12707)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12742..12761
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12821..12837)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12821..12837)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12834..12856)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12845..12861
                     /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(12869..12899)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12914..12935
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13052..13069)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13223..13811)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13303..13322)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(13982..14842)
                     /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     14605..14624
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14843..14947)
                     /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 aacaccggag acggttgtaa atgagcacac
     2881 aaaatacaca tgctaaaata ttatattcta tgacctttat aaaatcaacc aaaatcttct
     2941 ttttaataac tttagtatca ataattagaa tttttatgtt cctttttgca aacttttaat
     3001 aaaaatgagc aaaataaaaa aacgctagtt ttagtaactc gcgttgtttt cttcaccttt
     3061 aataatagct actccaccac ttgttcctaa gcggtcagct cctgcttcaa tcattttttg
     3121 agcatcttca aatgttctaa ctccaccagc tgctttaact aaagcattgt ctttaacaac
     3181 tgacttcatt agtttaacat cttcaaatgt tgcacctgat tttgaaaatc ctgttgatgt
     3241 tttaacaaat tctaatccag cttcaacagc tatttcacaa gctttcatga tttcttcttt
     3301 tgttaataaa caattttcca taatacattt aacaacatgt gatccagctg ctttttttac
     3361 agctttcatg tcttctaaaa ctaattcata atttttgtct tttaatgcac caatatttaa
     3421 taccatatca atttctgttg caccatcttt aattgcttca gaaacttcga atgcttttgt
     3481 agctgttgtg catgcaccta gaggaaaacc tacaacattt gttattccta catttgtgcc
     3541 ttttaataat tctttacaat agcttgttca atatgaatta acacaaactg ttgcaaaatc
     3601 aaattcaatt gcttcatcac ataattgttt aatttcagct ttcgtagcat cttgttttaa
     3661 taatgtgtga tctatatatt tgtttagttt cattttttct cctatatatt catttttaat
     3721 tttaattctt taataatttc gtctacttta actttagcgt tttgaacaga ttcaccaaca
     3781 cctataaaat aaatttttag tttaggttca gttccacttg ggcgaacagc aaatcatgac
     3841 ttatcttcta aataaaattt tagtaagtct tgtcctggca tattatacat tccatcgatg
     3901 tagtcttcaa cattaacaac tttaagtcca gcaatttgag ttaagggtgt tgctctcaat
     3961 gatttcatta atggttcaat ttttaatttc ttttcttctg gtttaaaatt caagtttaaa
     4021 gtgaaagtgt aatatgcacc catttcttta aataaatctt ctaaatagtc tactaatgtt
     4081 ttattttgtt ttttataaaa tcaagcagcc tctgctatta atatagaagc ttgtattcca
     4141 tctttatctc tagctgagtc atcaattaca tatccataac tttcttcata agcaaaaaca
     4201 aaatttaatc cgttatcttc ttctttagca atttctctac ccattcattt aaatccagtt
     4261 aaagttttta caatattaac tccatatttt tcatgagcga ttctatcacc caaatcactt
     4321 gttacaaaac ttgaatatag agccggattt tttggaatgc tatttaagcg ttttagattt
     4381 gataattttc aatcaattaa aattggtcct gtttgatttc catctaatct tacaaaatga
     4441 ccatcatgtt ttattgccat tccaaatctg tcagcatctg ggtcattcat aataataata
     4501 tctgcatcat gtttaatacc atattcaagc ggtatttttc atgcaggatc aaattctgga
     4561 tttggattta caacattttt aaatgtttca tcttcaaatg catgctcttc aacctcaata
     4621 acgttatatc ctgattcacg taatattttt ggggtaaatt tagttcctgt tccattaact
     4681 gcgctaaaaa taatttttaa atctttttta gcttcttgct cttttttgta cgtctctgtt
     4741 ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc
     4801 accgagtcgg tgcttttttg aattcgctag ctaggtcttg aaaggagtgg gaattggctc
     4861 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
     4921 ggtcggcaat tgatccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt
     4981 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt
     5041 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggacc ggttctagag
     5101 cgctgccacc atggacaaga agtacagcat cggcctggac atcggcacca actctgtggg
     5161 ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa
     5221 caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga
     5281 aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa
     5341 ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt
     5401 cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc
     5461 catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca
     5521 cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc
     5581 cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga
     5641 caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga
     5701 ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag
     5761 caagagcaga cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct
     5821 gttcggaaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga
     5881 cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa
     5941 cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc
     6001 cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct
     6061 gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc
     6121 tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa
     6181 cggctacgcc ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa
     6241 gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga
     6301 cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg
     6361 agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg
     6421 ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag
     6481 gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa
     6541 cttcgaggaa gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa
     6601 cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta
     6661 cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc
     6721 cgccttcctg agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg
     6781 gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc
     6841 cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct
     6901 gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga
     6961 agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa
     7021 aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac
     7081 cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa
     7141 gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat
     7201 ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg
     7261 cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat
     7321 cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga
     7381 gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag
     7441 ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa
     7501 agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca
     7561 gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga
     7621 tgtggaccat atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct
     7681 gaccagaagc gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa
     7741 gaagatgaag aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt
     7801 cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat
     7861 caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc
     7921 ccggatgaac actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac
     7981 cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga
     8041 gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct
     8101 gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga
     8161 cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt
     8221 cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat
     8281 ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt gggataaggg
     8341 ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa
     8401 gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaacagcga
     8461 taagctgatc gccagaaaga aggactggga ccctaagaag tacggcggct tcgacagccc
     8521 caccgtggcc tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact
     8581 gaagagtgtg aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa
     8641 tcccatcgac tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa
     8701 gctgcctaag tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc
     8761 cggcgaactg cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta
     8821 cctggccagc cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct
     8881 gtttgtggaa cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttctc
     8941 caagagagtg atcctggccg acgctaatct ggacaaagtg ctgtccgcct acaacaagca
     9001 ccgggataag cccatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa
     9061 tctgggagcc cctgccgcct tcaagtactt tgacaccacc atcgaccgga agaggtacac
     9121 cagcaccaaa gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga
     9181 gacacggatc gacctgtctc agctgggagg cgacaagcgt cctgctgcta ctaagaaagc
     9241 tggtcaagct aagaaaaaga aagctagcgg cagcggcgcc accaacttca gcctgctgaa
     9301 gcaggccggc gacgtggagg agaaccccgg ccccgttaac atggtgagca agggcgagga
     9361 ggtcatcaaa gagttcatgc gcttcaaggt gcgcatggag ggctccatga acggccacga
     9421 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     9481 gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc cccagttcat
     9541 gtacggctcc aaggcgtacg tgaagcaccc cgccgacatc cccgattaca agaagctgtc
     9601 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gtctggtgac
     9661 cgtgacccag gactcctccc tgcaggacgg cacgctgatc tacaaggtga agatgcgcgg
     9721 caccaacttc ccccccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     9781 caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccacc aggccctgaa
     9841 gctgaaggac ggcggccact acctggtgga gttcaagacc atctacatgg ccaagaagcc
     9901 cgtgcaactg cccggctact actacgtgga caccaagctg gacatcacct cccacaacga
     9961 ggactacacc atcgtggaac agtacgagcg ctccgagggc cgccaccacc tgttcctgta
    10021 cggcatggac gagctgtaca agtaaactag tacgcgttaa gtcgacaatc aacctctgga
    10081 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
    10141 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
    10201 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
    10261 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
    10321 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
    10381 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
    10441 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
    10501 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
    10561 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
    10621 gacgagtcgg atctcccttt gggccgcctc cccgcgtcga ctttaagacc aatgacttac
    10681 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
    10741 cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag
    10801 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
    10861 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
    10921 tccctcagac ccttttagtc agtgtggaaa atctctagca gggcccgttt aaacccgctg
    10981 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
    11041 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
    11101 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
    11161 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
    11221 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
    11281 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
    11341 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
    11401 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
    11461 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
    11521 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
    11581 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
    11641 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
    11701 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
    11761 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
    11821 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
    11881 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
    11941 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
    12001 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
    12061 ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat
    12121 aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt ccggtgctca
    12181 ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg
    12241 acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg
    12301 cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg
    12361 acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc
    12421 cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg
    12481 ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg
    12541 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
    12601 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
    12661 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
    12721 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
    12781 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
    12841 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
    12901 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
    12961 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
    13021 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
    13081 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
    13141 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
    13201 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
    13261 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
    13321 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
    13381 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
    13441 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
    13501 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
    13561 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
    13621 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
    13681 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
    13741 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
    13801 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
    13861 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
    13921 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
    13981 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
    14041 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
    14101 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
    14161 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
    14221 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
    14281 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
    14341 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
    14401 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
    14461 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
    14521 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
    14581 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
    14641 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
    14701 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
    14761 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
    14821 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
    14881 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
    14941 ttccgcgcac atttccccga aaagtgccac ctgac
//
