pC034-LwCas13a-msfGFP-2A-Blast

LOCUS       pC034-LwCas13a-msfGFP-2A-Blast                13090 bp ds-DNA     circular SYN 16-OCT-2019
DEFINITION  Expresses active LwCas13a for mammalian RNA targeting with 
            Blasticidin selection marker.
ACCESSION   .
VERSION     .
KEYWORDS    pC034 - LwCas13a-msfGFP-2A-Blast
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13090)
  AUTHORS   Abudayyeh OO, Gootenberg JS, Essletzbichler P, Han S, Joung J, 
            Belanto JJ, Verdine V, Cox DBT, Kellner MJ, Regev A, Lander ES, 
            Voytas DF, Ting AY, Zhang F
  TITLE     RNA targeting with CRISPR-Cas13.
  JOURNAL   Nature. 2017 Oct 4. doi: 10.1038/nature24049.
  PUBMED    28976959
REFERENCE   2  (bases 1 to 13090)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 16, 2019 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..13090
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1443..1676
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    2203..2320
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2413..2624
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2652..2699
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             2706..6161
                     /codon_start=1
                     /product="Leptotrichia wadei CRISPR-Cas effector that acts 
                     as an RNA-guided RNAse (Abudayyeh et al., 2017)"
                     /label=LwCas13a
                     /note="previously called C2c2"
                     /translation="MKVTKVDGISHKKYIEEGKLVKSTSEENRTSERLSELLSIRLDIY
                     IKNPDNASEEENRIRRENLKKFFSNKVLHLKDSVLYLKNRKEKNAVQDKNYSEEDISEY
                     DLKNKNSFSVLKKILLNEDVNSEELEIFRKDVEAKLNKINSLKYSFEENKANYQKINEN
                     NVEKVGGKSKRNIIYDYYRESAKRNDYINNVQEAFDKLYKKEDIEKLFFLIENSKKHEK
                     YKIREYYHKIIGRKNDKENFAKIIYEEIQNVNNIKELIEKIPDMSELKKSQVFYKYYLD
                     KEELNDKNIKYAFCHFVEIEMSQLLKNYVYKRLSNISNDKIKRIFEYQNLKKLIENKLL
                     NKLDTYVRNCGKYNYYLQVGEIATSDFIARNRQNEAFLRNIIGVSSVAYFSLRNILETE
                     NENGITGRMRGKTVKNNKGEEKYVSGEVDKIYNENKQNEVKENLKMFYSYDFNMDNKNE
                     IEDFFANIDEAISSIRHGIVHFNLELEGKDIFAFKNIAPSEISKKMFQNEINEKKLKLK
                     IFKQLNSANVFNYYEKDVIIKYLKNTKFNFVNKNIPFVPSFTKLYNKIEDLRNTLKFFW
                     SVPKDKEEKDAQIYLLKNIYYGEFLNKFVKNSKVFFKITNEVIKINKQRNQKTGHYKYQ
                     KFENIEKTVPVEYLAIIQSREMINNQDKEEKNTYIDFIQQIFLKGFIDYLNKNNLKYIE
                     SNNNNDNNDIFSKIKIKKDNKEKYDKILKNYEKHNRNKEIPHEINEFVREIKLGKILKY
                     TENLNMFYLILKLLNHKELTNLKGSLEKYQSANKEETFSDELELINLLNLDNNRVTEDF
                     ELEANEIGKFLDFNENKIKDRKELKKFDTNKIYFDGENIIKHRAFYNIKKYGMLNLLEK
                     IADKAKYKISLKELKEYSNKKNEIEKNYTMQQNLHRKYARPKKDEKFNDEDYKEYEKAI
                     GNIQKYTHLKNKVEFNELNLLQGLLLKILHRLVGYTSIWERDLRFRLKGEFPENHYIEE
                     IFNFDNSKNVKYKSGQIVEKYINFYKELYKDNVEKRSIYSDKKVKKLKQEKKDLYIRNY
                     IAHFNYIPHAEISLLEVLENLRKLLSYDRKLKNAIMKSIVDILKEYGFVATFKIGADKK
                     IEIQTLESEKIVHLKNLKKKKLMTDRNSEELCELVKVMFEYKALE"
     CDS             6927..6974
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6975..7055
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             7065..7121
                     /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             7122..7520
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    7545..8133
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7598..7618)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8016..8027)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8136..8152)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8137..8153)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             8658..8838
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8864..8881)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8870..9094
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9140..9568
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9227..9246)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9437..9458
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9577..9597)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9582..9911
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9762..9897
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9824..9843
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9959..10006
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10025..10399
                     /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    10529..10650
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10566..10585)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10620..10639
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10699..10715)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10699..10715)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10712..10734)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10723..10739
                     /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(10747..10777)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10792..10813
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10930..10947)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11101..11689)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11181..11200)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(11860..12720)
                     /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     12483..12502
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12721..12825)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12900..12919)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2341 cagagatcca gtttggttaa ttagctagct aggtcttgaa aggagtggga attggctccg
     2401 gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg
     2461 tcggcaattg atccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg
     2521 tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg
     2581 ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggaccgg ttctagagcg
     2641 ctgccaccat gaagcgacct gccgccacaa agaaggctgg acaggctaag aagaagaaag
     2701 gatccatgaa agtgaccaag gtcgacggca tcagccacaa gaagtacatc gaagagggca
     2761 agctcgtgaa gtccaccagc gaggaaaacc ggaccagcga gagactgagc gagctgctga
     2821 gcatccggct ggacatctac atcaagaacc ccgacaacgc ctccgaggaa gagaaccgga
     2881 tcagaagaga gaacctgaag aagttcttta gcaacaaggt gctgcacctg aaggacagcg
     2941 tgctgtatct gaagaaccgg aaagaaaaga acgccgtgca ggacaagaac tatagcgaag
     3001 aggacatcag cgagtacgac ctgaaaaaca agaacagctt ctccgtgctg aagaagatcc
     3061 tgctgaacga ggacgtgaac tctgaggaac tggaaatctt tcggaaggac gtggaagcca
     3121 agctgaacaa gatcaacagc ctgaagtaca gcttcgaaga gaacaaggcc aactaccaga
     3181 agatcaacga gaacaacgtg gaaaaagtgg gcggcaagag caagcggaac atcatctacg
     3241 actactacag agagagcgcc aagcgcaacg actacatcaa caacgtgcag gaagccttcg
     3301 acaagctgta taagaaagag gatatcgaga aactgttttt cctgatcgag aacagcaaga
     3361 agcacgagaa gtacaagatc cgcgagtact atcacaagat catcggccgg aagaacgaca
     3421 aagagaactt cgccaagatt atctacgaag agatccagaa cgtgaacaac atcaaagagc
     3481 tgattgagaa gatccccgac atgtctgagc tgaagaaaag ccaggtgttc tacaagtact
     3541 acctggacaa agaggaactg aacgacaaga atattaagta cgccttctgc cacttcgtgg
     3601 aaatcgagat gtcccagctg ctgaaaaact acgtgtacaa gcggctgagc aacatcagca
     3661 acgataagat caagcggatc ttcgagtacc agaatctgaa aaagctgatc gaaaacaaac
     3721 tgctgaacaa gctggacacc tacgtgcgga actgcggcaa gtacaactac tatctgcaag
     3781 tgggcgagat cgccacctcc gactttatcg cccggaaccg gcagaacgag gccttcctga
     3841 gaaacatcat cggcgtgtcc agcgtggcct acttcagcct gaggaacatc ctggaaaccg
     3901 agaacgagaa cggtatcacc ggccggatgc ggggcaagac cgtgaagaac aacaagggcg
     3961 aagagaaata cgtgtccggc gaggtggaca agatctacaa tgagaacaag cagaacgaag
     4021 tgaaagaaaa tctgaagatg ttctacagct acgacttcaa catggacaac aagaacgaga
     4081 tcgaggactt cttcgccaac atcgacgagg ccatcagcag catcagacac ggcatcgtgc
     4141 acttcaacct ggaactggaa ggcaaggaca tcttcgcctt caagaatatc gcccccagcg
     4201 agatctccaa gaagatgttt cagaacgaaa tcaacgaaaa gaagctgaag ctgaaaatct
     4261 tcaagcagct gaacagcgcc aacgtgttca actactacga gaaggatgtg atcatcaagt
     4321 acctgaagaa taccaagttc aacttcgtga acaaaaacat ccccttcgtg cccagcttca
     4381 ccaagctgta caacaagatt gaggacctgc ggaataccct gaagtttttt tggagcgtgc
     4441 ccaaggacaa agaagagaag gacgcccaga tctacctgct gaagaatatc tactacggcg
     4501 agttcctgaa caagttcgtg aaaaactcca aggtgttctt taagatcacc aatgaagtga
     4561 tcaagattaa caagcagcgg aaccagaaaa ccggccacta caagtatcag aagttcgaga
     4621 acatcgagaa aaccgtgccc gtggaatacc tggccatcat ccagagcaga gagatgatca
     4681 acaaccagga caaagaggaa aagaatacct acatcgactt tattcagcag attttcctga
     4741 agggcttcat cgactacctg aacaagaaca atctgaagta tatcgagagc aacaacaaca
     4801 atgacaacaa cgacatcttc tccaagatca agatcaaaaa ggataacaaa gagaagtacg
     4861 acaagatcct gaagaactat gagaagcaca atcggaacaa agaaatccct cacgagatca
     4921 atgagttcgt gcgcgagatc aagctgggga agattctgaa gtacaccgag aatctgaaca
     4981 tgttttacct gatcctgaag ctgctgaacc acaaagagct gaccaacctg aagggcagcc
     5041 tggaaaagta ccagtccgcc aacaaagaag aaaccttcag cgacgagttg gaactgatca
     5101 acctgctgaa cctggacaac aacagagtga ccgaggactt cgagctggaa gccaacgaga
     5161 tcggcaagtt cctggacttc aacgaaaaca aaatcaagga ccggaaagag ctgaaaaagt
     5221 tcgacaccaa caagatctat ttcgacggcg agaacatcat caagcaccgg gccttctaca
     5281 atatcaagaa atacggcatg ctgaatctgc tggaaaagat cgccgataag gccaagtata
     5341 agatcagcct gaaagaactg aaagagtaca gcaacaagaa gaatgagatt gaaaagaact
     5401 acaccatgca gcagaacctg caccggaagt acgccagacc caagaaggac gaaaagttca
     5461 acgacgagga ctacaaagag tatgagaagg ccatcggcaa catccagaag tacacccacc
     5521 tgaagaacaa ggtggaattc aatgagctga acctgctgca gggcctgctg ctgaagatcc
     5581 tgcaccggct cgtgggctac accagcatct gggagcggga cctgagattc cggctgaagg
     5641 gcgagtttcc cgagaaccac tacatcgagg aaattttcaa tttcgacaac tccaagaatg
     5701 tgaagtacaa aagcggccag atcgtggaaa agtatatcaa cttctacaaa gaactgtaca
     5761 aggacaatgt ggaaaagcgg agcatctact ccgacaagaa agtgaagaaa ctgaagcagg
     5821 aaaaaaagga cctgtacatc cggaactaca ttgcccactt caactacatc ccccacgccg
     5881 agattagcct gctggaagtg ctggaaaacc tgcggaagct gctgtcctac gaccggaagc
     5941 tgaagaacgc catcatgaag tccatcgtgg acattctgaa agaatacggc ttcgtggcca
     6001 ccttcaagat cggcgctgac aagaagatcg aaatccagac cctggaatca gagaagatcg
     6061 tgcacctgaa gaatctgaag aaaaagaaac tgatgaccga ccggaacagc gaggaactgt
     6121 gcgaactcgt gaaagtcatg ttcgagtaca aggccctgga aggaggaggt ggaagcggag
     6181 gaggaggaag cggaggagga ggtagcgtga gtaaaggtga agaactcttc actggagtag
     6241 tgcccattct ggtagagctt gatggagatg taaatggaca taaattctcc gtcaggggcg
     6301 aaggcgaagg ggacgccacg aatggtaagc tgactctgaa attcatctgt acgacgggca
     6361 aactgcccgt cccatggcct acactcgtaa cgaccctcac ctacggcgtg caatgctttt
     6421 ctcgatatcc cgaccacatg aaacagcatg actttttcaa gtctgcaatg cctgaaggtt
     6481 atgttcaaga aaggaccatc agctttaagg atgatggtac atataaaacc cgagccgagg
     6541 ttaaatttga aggggacact ctggttaatc gaattgaact gaaaggtatt gattttaagg
     6601 aggacggtaa catactgggg cacaagttgg agtacaactt taacagccat aatgtgtata
     6661 ttaccgctga taagcagaaa aatgggataa aggccaactt taagatccga cataatgtcg
     6721 aagatggtag tgttcaactg gctgatcatt accaacaaaa tacgcccatc ggagatggac
     6781 ctgtactctt gcctgacaat cattatctct ccacgcaatc aaagctttcc aaggacccaa
     6841 acgaaaagag agatcacatg gtccttctgg aatttgtgac tgccgcaggc atcactctcg
     6901 gtatggatga gctgtacaag ggatccaagc gacctgccgc cacaaagaag gctggacagg
     6961 ctaagaagaa gaaataccca tacgatgttc cagattacgc ttatccctac gacgtgcctg
     7021 attatgcata cccatatgat gtccccgact atgccggatc cggcgcaaca aacttctctc
     7081 tgctgaaaca agccggagat gtcgaagaga atcctggacc gatggccaag cctttgtctc
     7141 aagaagaatc caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg
     7201 aagactacag cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca
     7261 atgtatatca ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg
     7321 ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct
     7381 tgagcccctg cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca
     7441 tagtgaagga cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg
     7501 gttatgtgtg ggagggctaa gaattcgata tcaagcttat cggtaatcaa cctctggatt
     7561 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     7621 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     7681 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     7741 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     7801 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     7861 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     7921 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
     7981 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     8041 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     8101 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgacct cgagacctag
     8161 aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc
     8221 tagaagcaca agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac
     8281 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     8341 aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc taccacacac
     8401 aaggctactt ccctgattgg cagaactaca caccagggcc agggatcaga tatccactga
     8461 cctttggatg gtgctacaag ctagtaccag ttgagcaaga gaaggtagaa gaagccaatg
     8521 aaggagagaa cacccgcttg ttacaccctg tgagcctgca tgggatggat gacccggaga
     8581 gagaagtatt agagtggagg tttgacagcc gcctagcatt tcatcacatg gcccgagagc
     8641 tgcatccgga ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     8701 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     8761 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     8821 tgtggaaaat ctctagcagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     8881 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     8941 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     9001 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     9061 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     9121 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     9181 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     9241 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     9301 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     9361 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     9421 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     9481 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     9541 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     9601 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     9661 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     9721 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     9781 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     9841 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     9901 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt
     9961 tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta
    10021 aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc
    10081 ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc
    10141 cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc
    10201 ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc
    10261 ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga
    10321 gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt
    10381 ggccgaggag caggactgac acgtgctacg agatttcgat tccaccgccg ccttctatga
    10441 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
    10501 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
    10561 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
    10621 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
    10681 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
    10741 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
    10801 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
    10861 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
    10921 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
    10981 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
    11041 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
    11101 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
    11161 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
    11221 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
    11281 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
    11341 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
    11401 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
    11461 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
    11521 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
    11581 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
    11641 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
    11701 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
    11761 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
    11821 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
    11881 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
    11941 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
    12001 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
    12061 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
    12121 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
    12181 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
    12241 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
    12301 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
    12361 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
    12421 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
    12481 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
    12541 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
    12601 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
    12661 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
    12721 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
    12781 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
    12841 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
    12901 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
    12961 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
    13021 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
    13081 tatacgcgtt
//