LOCUS       Exported               16811 bp ds-DNA     circular SYN 18-JAN-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16811)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Apr 30, 2019 from SnapGene Viewer 4.3.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..16811
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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"
     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."
     misc_feature    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    2599..2632
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        2647..2887
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_RNA        4777..4852
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4889..5203
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    4897..4915
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4933..4951
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4968..4986
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5004..5022
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5040..5058
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5075..5093
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5111..5129
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             5235..9338
                     /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             9339..9386
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9387..9410
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             9420..9473
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             9474..10190
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     promoter        10201..10412
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             10434..11030
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     CDS             11040..11096
                     /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             11097..11843
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     protein_bind    11850..11883
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    11903..12491
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(12374..12385)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             12563..12796
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    12828..13052
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      13098..13526
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        13540..13869
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      13720..13855
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        13917..13964
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             13983..14357
                     /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    14487..14608
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(14657..14673)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    14681..14697
                     /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(14705..14735)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14750..14771
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(15059..15647)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(15818..16678)
                     /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"
     promoter        complement(16679..16783)
                     /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 tggttaatat aacttcgtat agcatacatt atacgaagtt atttaattaa
     2641 ggtaccgagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
     2701 agagagataa ttagaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
     2761 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
     2821 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
     2881 aaaggacgaa acaccggaga cggttgtaaa tgagcacaca aaatacacat gctaaaatat
     2941 tatattctat gacctttata aaatcaacca aaatcttctt tttaataact ttagtatcaa
     3001 taattagaat ttttatgttc ctttttgcaa acttttaata aaaatgagca aaataaaaaa
     3061 acgctagttt tagtaactcg cgttgttttc ttcaccttta ataatagcta ctccaccact
     3121 tgttcctaag cggtcagctc ctgcttcaat cattttttga gcatcttcaa atgttctaac
     3181 tccaccagct gctttaacta aagcattgtc tttaacaact gacttcatta gtttaacatc
     3241 ttcaaatgtt gcacctgatt ttgaaaatcc tgttgatgtt ttaacaaatt ctaatccagc
     3301 ttcaacagct atttcacaag ctttcatgat ttcttctttt gttaataaac aattttccat
     3361 aatacattta acaacatgtg atccagctgc tttttttaca gctttcatgt cttctaaaac
     3421 taattcataa tttttgtctt ttaatgcacc aatatttaat accatatcaa tttctgttgc
     3481 accatcttta attgcttcag aaacttcgaa tgcttttgta gctgttgtgc atgcacctag
     3541 aggaaaacct acaacatttg ttattcctac atttgtgcct tttaataatt ctttacaata
     3601 gcttgttcaa tatgaattaa cacaaactgt tgcaaaatca aattcaattg cttcatcaca
     3661 taattgttta atttcagctt tcgtagcatc ttgttttaat aatgtgtgat ctatatattt
     3721 gtttagtttc attttttctc ctatatattc atttttaatt ttaattcttt aataatttcg
     3781 tctactttaa ctttagcgtt ttgaacagat tcaccaacac ctataaaata aatttttagt
     3841 ttaggttcag ttccacttgg gcgaacagca aatcatgact tatcttctaa ataaaatttt
     3901 agtaagtctt gtcctggcat attatacatt ccatcgatgt agtcttcaac attaacaact
     3961 ttaagtccag caatttgagt taagggtgtt gctctcaatg atttcattaa tggttcaatt
     4021 tttaatttct tttcttctgg tttaaaattc aagtttaaag tgaaagtgta atatgcaccc
     4081 atttctttaa ataaatcttc taaatagtct actaatgttt tattttgttt tttataaaat
     4141 caagcagcct ctgctattaa tatagaagct tgtattccat ctttatctct agctgagtca
     4201 tcaattacat atccataact ttcttcataa gcaaaaacaa aatttaatcc gttatcttct
     4261 tctttagcaa tttctctacc cattcattta aatccagtta aagtttttac aatattaact
     4321 ccatattttt catgagcgat tctatcaccc aaatcacttg ttacaaaact tgaatataga
     4381 gccggatttt ttggaatgct atttaagcgt tttagatttg ataattttca atcaattaaa
     4441 attggtcctg tttgatttcc atctaatctt acaaaatgac catcatgttt tattgccatt
     4501 ccaaatctgt cagcatctgg gtcattcata ataataatat ctgcatcatg tttaatacca
     4561 tattcaagcg gtatttttca tgcaggatca aattctggat ttggatttac aacattttta
     4621 aatgtttcat cttcaaatgc atgctcttca acctcaataa cgttatatcc tgattcacgt
     4681 aatatttttg gggtaaattt agttcctgtt ccattaactg cgctaaaaat aatttttaaa
     4741 tcttttttag cttcttgctc ttttttgtac gtctctgttt tagagctaga aatagcaagt
     4801 taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcttttttga
     4861 attcactttg gccgcgaatc gatatgtcga gtttactccc tatcagtgat agagaacgta
     4921 tgtcgagttt actccctatc agtgatagag aacgatgtcg agtttactcc ctatcagtga
     4981 tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgtatgt cgagtttact
     5041 ccctatcagt gatagagaac gtatgtcgag tttatcccta tcagtgatag agaacgtatg
     5101 tcgagtttac tccctatcag tgatagagaa cgtatgtcga ggtaggcgtg tacggtggga
     5161 ggcctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagaa taccggttct
     5221 agagcgctgc caccatggac aagaagtaca gcatcggcct ggacatcggc accaactctg
     5281 tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg
     5341 gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg
     5401 gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga
     5461 agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca
     5521 gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc
     5581 accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct
     5641 accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc
     5701 tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc
     5761 ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt
     5821 tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac
     5881 tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg
     5941 gcctgttcgg aaacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact
     6001 tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg
     6061 acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc
     6121 tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc
     6181 ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga
     6241 aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca
     6301 agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca
     6361 tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag
     6421 aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc
     6481 tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca
     6541 accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg
     6601 ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct
     6661 ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga
     6721 ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg
     6781 agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa
     6841 agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca
     6901 accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg
     6961 actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg
     7021 atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc
     7081 tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc
     7141 tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat
     7201 acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg
     7261 gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc
     7321 tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc
     7381 agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg
     7441 gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc
     7501 ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga
     7561 acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc
     7621 tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc
     7681 tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact
     7741 acgatgtgga ccatatcgtg cctcagagct ttctgaagga cgactccatc gacaacaagg
     7801 tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg
     7861 tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa
     7921 agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct
     7981 tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg
     8041 actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga
     8101 tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc
     8161 gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg
     8221 ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt
     8281 acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt
     8341 acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg
     8401 agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata
     8461 agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga
     8521 aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca
     8581 gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca
     8641 gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga
     8701 aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga
     8761 agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca
     8821 tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct
     8881 ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc
     8941 tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac
     9001 agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt
     9061 tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca
     9121 agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga
     9181 ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt
     9241 acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt
     9301 acgagacacg gatcgacctg tctcagctgg gaggcgacaa gcgacctgcc gccacaaaga
     9361 aggctggaca ggctaagaag aagaaagatt acaaagacga tgacgataag ggatccggag
     9421 agggcagagg aagtctgcta acatgcggtg acgtcgagga gaatcctggc ccagtgagca
     9481 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     9541 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     9601 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     9661 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     9721 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     9781 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     9841 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     9901 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     9961 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
    10021 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
    10081 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
    10141 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa gctagcatcc
    10201 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgat
    10261 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
    10321 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
    10381 tttttcgcaa cgggtttgcc gccagaacac agactgcgat cgcaatgtac agtatgaccg
    10441 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
    10501 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
    10561 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
    10621 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
    10681 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
    10741 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
    10801 ggttcctggc caccgtcgga gtctcgcccg accaccaggg caagggtctg ggcagcgccg
    10861 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
    10921 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
    10981 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc ggttccggcg
    11041 caacaaactt ctctctgctg aaacaagccg gagatgtcga agagaatcct ggaccgatgt
    11101 ctagactgga caagagcaaa gtcataaacg gagctctgga attactcaat ggtgtcggta
    11161 tcgaaggcct gacgacaagg aaactcgctc aaaagctggg agttgagcag cctaccctgt
    11221 actggcacgt gaagaacaag cgggccctgc tcgatgccct gccaatcgag atgctggaca
    11281 ggcatcatac ccacttctgc cccctggaag gcgagtcatg gcaagacttt ctgcggaaca
    11341 acgccaagtc ataccgctgt gctctcctct cacatcgcga cggggctaaa gtgcatctcg
    11401 gcacccgccc aacagagaaa cagtacgaaa ccctggaaaa tcagctcgcg ttcctgtgtc
    11461 agcaaggctt ctccctggag aacgcactgt acgctctgtc cgccgtgggc cactttacac
    11521 tgggctgcgt attggaggaa caggagcatc aagtagcaaa agaggaaaga gagacaccta
    11581 ccaccgattc tatgccccca cttctgagac aagcaattga gctgttcgac cggcagggag
    11641 ccgaacctgc cttccttttc ggcctggaac taatcatatg tggcctggag aaacagctaa
    11701 agtgcgaaag cggcgggccg accgacgccc ttgacgattt tgacttagac atgctcccag
    11761 ccgatgccct tgacgacttt gaccttgata tgctgcctgc tgacgctctt gacgattttg
    11821 accttgacat gctccccggg taactcgaga taacttcgta tagcatacat tatacgaagt
    11881 tattcgagcg cgttaagtcg acaatcaacc tctggattac aaaatttgtg aaagattgac
    11941 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
    12001 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
    12061 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
    12121 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
    12181 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
    12241 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
    12301 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
    12361 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
    12421 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
    12481 cgcctccccg cgtcgacttt aagaccaatg acttacaagg cagctgtaga tcttagccac
    12541 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg
    12601 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
    12661 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
    12721 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
    12781 tggaaaatct ctagcagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
    12841 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
    12901 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
    12961 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
    13021 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
    13081 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
    13141 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
    13201 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
    13261 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
    13321 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
    13381 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
    13441 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
    13501 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
    13561 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
    13621 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
    13681 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
    13741 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
    13801 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
    13861 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg
    13921 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
    13981 ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg
    14041 tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg
    14101 gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg
    14161 acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg
    14221 aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc
    14281 agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg
    14341 ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa
    14401 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
    14461 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
    14521 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
    14581 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
    14641 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
    14701 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
    14761 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
    14821 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
    14881 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
    14941 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
    15001 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
    15061 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
    15121 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
    15181 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
    15241 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
    15301 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
    15361 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
    15421 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
    15481 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
    15541 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
    15601 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
    15661 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
    15721 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
    15781 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
    15841 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
    15901 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
    15961 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
    16021 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
    16081 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
    16141 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
    16201 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
    16261 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
    16321 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
    16381 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
    16441 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
    16501 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
    16561 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
    16621 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
    16681 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
    16741 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
    16801 tgccacctga c
//
