LOCUS       sequence-382455-       12838 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses human codon-optimized Sniper2L and blasticidin resistance:
            EFS promoter-Sniper2L-NLS-FLAG-P2A-BSD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12838)
  AUTHORS   Kim YH, Kim N, Okafor I, Choi S, Min S, Lee J, Bae SM, Choi K, Choi 
            J, Harihar V, Kim Y, Kim JS, Kleinstiver BP, Lee JK, Ha T, Kim HH
  TITLE     Sniper2L is a high-fidelity Cas9 variant with high activity.
  JOURNAL   Nat Chem Biol. 2023 Mar 9. doi: 10.1038/s41589-023-01279-5.
  PUBMED    36894722
REFERENCE   2  (bases 1 to 12838)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Chem 
            Biol. 2023 Mar 9. doi: 10.1038/s41589-023-01279-5."
COMMENT     Sequence Label: Sniper2L
FEATURES             Location/Qualifiers
     source          1..12838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    5..26
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(143..160)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(314..902)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(394..413)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1073..1864,1865..1933))
                     /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     1696..1715
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1934..2038)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2113..2132)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2304..2683
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2685..2883
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2838..2858
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2901..3081
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3128..3253
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3746..3979
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4164..4208
                     /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    4506..4623
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4716..4927
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      4946..4955
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4952..9055
                     /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
                     FTVYNELTKVKYVTEGMRKPASLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEIARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNANLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESLFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             9056..9103
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9104..9127
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             9137..9193
                     /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             9194..9592
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    9617..10205
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9670..9690)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10088..10099)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10208..10224)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10209..10225)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10730..10910
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10936..10953)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10942..11166
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11212..11640
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11299..11318)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11509..11530
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11649..11669)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      11814..11948
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11875..11894
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12010..12057
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12076..12450
                     /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    12580..12701
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12617..12636)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12671..12690
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12750..12766)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12750..12766)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12763..12785)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12774..12790
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(12798..12804,12805..12822,12823..12828))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
       61 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
      121 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
      181 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
      241 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
      301 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
      361 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
      421 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
      481 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
      541 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
      601 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
      661 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
      721 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
      781 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
      841 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
      901 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
      961 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     1021 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     1081 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     1141 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     1201 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     1261 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     1321 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     1381 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     1441 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     1501 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     1561 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     1621 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     1681 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     1741 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     1801 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     1861 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     1921 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     1981 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     2041 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     2101 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     2161 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     2221 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
     2281 tgtacgggcc agatatacgc gttgacattg attattgact agttattaat agtaatcaat
     2341 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     2401 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     2461 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     2521 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     2581 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     2641 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
     2701 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     2761 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     2821 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     2881 cagcgcgttt tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     2941 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     3001 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     3061 cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc
     3121 agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg
     3181 gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg
     3241 gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt
     3301 aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct
     3361 agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact
     3421 gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac
     3481 agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt
     3541 agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga
     3601 tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata
     3661 aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc
     3721 agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag
     3781 gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg
     3841 gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc
     3901 aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc
     3961 taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg
     4021 ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga
     4081 cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg
     4141 aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg
     4201 caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa
     4261 tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata
     4321 gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac
     4381 ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc
     4441 gattagtgaa cggatcggca ctgcgtgcgc caattctgca gacaaatggc agtattcatc
     4501 cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac
     4561 ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat
     4621 tttcgggttt attacaggga cagcagagat ccagtttggt taattagcta gctaggtctt
     4681 gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
     4741 ccccgagaag ttggggggag gggtcggcaa ttgatccggt gcctagagaa ggtggcgcgg
     4801 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
     4861 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
     4921 aacacaggac cggttctaga gcgctgccac catggacaag aagtacagca tcggcctgga
     4981 catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa
     5041 gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc
     5101 cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag
     5161 aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat
     5221 ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga
     5281 taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga
     5341 gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga
     5401 cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat
     5461 cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca
     5521 gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc
     5581 catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc
     5641 cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc
     5701 caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac
     5761 ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt
     5821 tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac
     5881 cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca
     5941 ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat
     6001 tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga
     6061 agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct
     6121 cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat
     6181 cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta
     6241 cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta
     6301 ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga
     6361 ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag
     6421 cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa
     6481 gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt
     6541 gaccgaggga atgagaaagc ccgccagcct gagcggcgag cagaaaaagg ccatcgtgga
     6601 cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa
     6661 gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc
     6721 cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga
     6781 ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga
     6841 gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca
     6901 gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat
     6961 ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa
     7021 cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa
     7081 agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag
     7141 ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt
     7201 gatgggccgg cacaagcccg agaacatcgt gatcgaaatc gccagagaga accagaccac
     7261 ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga
     7321 gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa
     7381 gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat
     7441 caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc tgaaggacga
     7501 ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt
     7561 gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa
     7621 cctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga
     7681 actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca
     7741 cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat
     7801 ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt
     7861 ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa
     7921 cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcc tgttcgtgta
     7981 cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg
     8041 caaggctacc gccaagtact tcttctacag caacatcatg aactttttca agaccgagat
     8101 taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg
     8161 ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc
     8221 ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat
     8281 cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa
     8341 gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga
     8401 aaagggcaag tccaagaaac tgaagagtgt gaaagagcta ctggggatca ccatcatgga
     8461 aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt
     8521 gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg
     8581 gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc
     8641 caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga
     8701 ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat
     8761 cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt
     8821 gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat
     8881 ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac
     8941 catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca
     9001 gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag gcgacaagcg
     9061 acctgccgcc acaaagaagg ctggacaggc taagaagaag aaagattaca aagacgatga
     9121 cgataaggga tccggcgcaa caaacttctc tctgctgaaa caagccggag atgtcgaaga
     9181 gaatcctgga ccgatggcca agcctttgtc tcaagaagaa tccaccctca ttgaaagagc
     9241 aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct
     9301 ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg
     9361 tgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat
     9421 cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt
     9481 gcttctcgat ctgcatcctg ggatcaaagc catagtgaag gacagtgatg gacagccgac
     9541 ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggct aagaattcga
     9601 tatcaagctt atcggtaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     9661 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     9721 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     9781 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     9841 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     9901 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     9961 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
    10021 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
    10081 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
    10141 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
    10201 cccgcatcga taccgtcgac ctcgagacct agaaaaacat ggagcaatca caagtagcaa
    10261 tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg
    10321 ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct
    10381 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
    10441 agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt ggcagaacta
    10501 cacaccaggg ccagggatca gatatccact gacctttgga tggtgctaca agctagtacc
    10561 agttgagcaa gagaaggtag aagaagccaa tgaaggagag aacacccgct tgttacaccc
    10621 tgtgagcctg catgggatgg atgacccgga gagagaagta ttagagtgga ggtttgacag
    10681 ccgcctagca tttcatcaca tggcccgaga gctgcatccg gactgtactg ggtctctctg
    10741 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
    10801 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
    10861 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gggcccgttt
    10921 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
    10981 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
    11041 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
    11101 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
    11161 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
    11221 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
    11281 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
    11341 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
    11401 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
    11461 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
    11521 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
    11581 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
    11641 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
    11701 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
    11761 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
    11821 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
    11881 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
    11941 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
    12001 tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
    12061 gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac
    12121 gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag
    12181 gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac
    12241 caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac
    12301 gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc
    12361 gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc
    12421 gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc
    12481 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
    12541 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
    12601 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
    12661 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
    12721 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
    12781 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtg
//
