LOCUS       sequence-382456-       12838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human codon-optimized Sniper2P and blasticidin resistance:
            EFS promoter-Sniper2P-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 Dec 15, 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: Sniper2P
FEATURES             Location/Qualifiers
     source          1..12838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(21..38)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(192..780)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(272..291)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(951..1742,1743..1811))
                     /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     1574..1593
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1812..1916)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1991..2010)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2182..2561
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2563..2761
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2716..2736
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2779..2959
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3006..3131
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3624..3857
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4042..4086
                     /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    4384..4501
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4594..4805
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      4824..4833
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4830..8933
                     /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
                     NAVVGTALIKKYPKLESPFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             8934..8981
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8982..9005
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             9015..9071
                     /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             9072..9470
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    9495..10083
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9548..9568)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9966..9977)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10086..10102)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10087..10103)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10608..10788
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10814..10831)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10820..11044
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11090..11518
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11177..11196)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11387..11408
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11527..11547)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      11692..11826
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11753..11772
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11888..11935
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11954..12328
                     /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    12458..12579
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12495..12514)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12549..12568
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12628..12644)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12628..12644)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12641..12663)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12652..12668
                     /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(12676..12682,12683..12700,12701..12706))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12721..12742
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
       61 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
      121 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
      181 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
      241 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
      301 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
      361 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
      421 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
      481 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
      541 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
      601 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
      661 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
      721 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
      781 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
      841 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      901 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      961 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     1021 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     1081 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     1141 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     1201 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     1261 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     1321 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     1381 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     1441 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     1501 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     1561 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     1621 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     1681 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     1741 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1801 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1861 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1921 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     1981 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     2041 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     2101 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
     2161 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
     2221 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     2281 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     2341 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     2401 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     2461 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     2521 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
     2581 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     2641 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     2701 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     2761 gcgcgttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     2821 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     2881 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     2941 gtgtggaaaa tctctagcag tggcgcccga acagggactt gaaagcgaaa gggaaaccag
     3001 aggagctctc tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc
     3061 ggcgactggt gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt
     3121 gcgagagcgt cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa
     3181 ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag
     3241 aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg
     3301 gacagctaca accatccctt cagacaggat cagaagaact tagatcatta tataatacag
     3361 tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag
     3421 acaagataga ggaagagcaa aacaaaagta agaccaccgc acagcaagcg gccgctgatc
     3481 ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa
     3541 gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag
     3601 agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga
     3661 agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt
     3721 atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa
     3781 ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta
     3841 aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct
     3901 gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc
     3961 tggatggagt gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa
     4021 gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca
     4081 agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg
     4141 atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga
     4201 gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc
     4261 gacaggcccg aaggaataga agaagaaggt ggagagagag acagagacag atccattcga
     4321 ttagtgaacg gatcggcact gcgtgcgcca attctgcaga caaatggcag tattcatcca
     4381 caattttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     4441 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     4501 tcgggtttat tacagggaca gcagagatcc agtttggtta attagctagc taggtcttga
     4561 aaggagtggg aattggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc
     4621 ccgagaagtt ggggggaggg gtcggcaatt gatccggtgc ctagagaagg tggcgcgggg
     4681 taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac
     4741 cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa
     4801 cacaggaccg gttctagagc gctgccacca tggacaagaa gtacagcatc ggcctggaca
     4861 tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg cccagcaaga
     4921 aattcaaggt gctgggcaac accgaccggc acagcatcaa gaagaacctg atcggagccc
     4981 tgctgttcga cagcggcgaa acagccgagg ccacccggct gaagagaacc gccagaagaa
     5041 gatacaccag acggaagaac cggatctgct atctgcaaga gatcttcagc aacgagatgg
     5101 ccaaggtgga cgacagcttc ttccacagac tggaagagtc cttcctggtg gaagaggata
     5161 agaagcacga gcggcacccc atcttcggca acatcgtgga cgaggtggcc taccacgaga
     5221 agtaccccac catctaccac ctgagaaaga aactggtgga cagcaccgac aaggccgacc
     5281 tgcggctgat ctatctggcc ctggcccaca tgatcaagtt ccggggccac ttcctgatcg
     5341 agggcgacct gaaccccgac aacagcgacg tggacaagct gttcatccag ctggtgcaga
     5401 cctacaacca gctgttcgag gaaaacccca tcaacgccag cggcgtggac gccaaggcca
     5461 tcctgtctgc cagactgagc aagagcagac ggctggaaaa tctgatcgcc cagctgcccg
     5521 gcgagaagaa gaatggcctg ttcggaaacc tgattgccct gagcctgggc ctgaccccca
     5581 acttcaagag caacttcgac ctggccgagg atgccaaact gcagctgagc aaggacacct
     5641 acgacgacga cctggacaac ctgctggccc agatcggcga ccagtacgcc gacctgtttc
     5701 tggccgccaa gaacctgtcc gacgccatcc tgctgagcga catcctgaga gtgaacaccg
     5761 agatcaccaa ggcccccctg agcgcctcta tgatcaagag atacgacgag caccaccagg
     5821 acctgaccct gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac aaagagattt
     5881 tcttcgacca gagcaagaac ggctacgccg gctacattga cggcggagcc agccaggaag
     5941 agttctacaa gttcatcaag cccatcctgg aaaagatgga cggcaccgag gaactgctcg
     6001 tgaagctgaa cagagaggac ctgctgcgga agcagcggac cttcgacaac ggcagcatcc
     6061 cccaccagat ccacctggga gagctgcacg ccattctgcg gcggcaggaa gatttttacc
     6121 cattcctgaa ggacaaccgg gaaaagatcg agaagatcct gaccttccgc atcccctact
     6181 acgtgggccc tctggccagg ggaaacagca gattcgcctg gatgaccaga aagagcgagg
     6241 aaaccatcac cccctggaac ttcgaggaag tggtggacaa gggcgcttcc gcccagagct
     6301 tcatcgagcg gatgaccaac ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc
     6361 acagcctgct gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga
     6421 ccgagggaat gagaaagccc gccagcctga gcggcgagca gaaaaaggcc atcgtggacc
     6481 tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac tacttcaaga
     6541 aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga agatcggttc aacgcctccc
     6601 tgggcacata ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg
     6661 aaaacgagga cattctggaa gatatcgtgc tgaccctgac actgtttgag gacagagaga
     6721 tgatcgagga acggctgaaa acctatgccc acctgttcga cgacaaagtg atgaagcagc
     6781 tgaagcggcg gagatacacc ggctggggca ggctgagccg gaagctgatc aacggcatcc
     6841 gggacaagca gtccggcaag acaatcctgg atttcctgaa gtccgacggc ttcgccaaca
     6901 gaaacttcat gcagctgatc cacgacgaca gcctgacctt taaagaggac atccagaaag
     6961 cccaggtgtc cggccagggc gatagcctgc acgagcacat tgccaatctg gccggcagcc
     7021 ccgccattaa gaagggcatc ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga
     7081 tgggccggca caagcccgag aacatcgtga tcgaaatcgc cagagagaac cagaccaccc
     7141 agaagggaca gaagaacagc cgcgagagaa tgaagcggat cgaagagggc atcaaagagc
     7201 tgggcagcca gatcctgaaa gaacaccccg tggaaaacac ccagctgcag aacgagaagc
     7261 tgtacctgta ctacctgcag aatgggcggg atatgtacgt ggaccaggaa ctggacatca
     7321 accggctgtc cgactacgat gtggaccata tcgtgcctca gagctttctg aaggacgact
     7381 ccatcgacaa caaggtgctg accagaagcg acaagaaccg gggcaagagc gacaacgtgc
     7441 cctccgaaga ggtcgtgaag aagatgaaga actactggcg gcagctgctg aacgccaacc
     7501 tgattaccca gagaaagttc gacaatctga ccaaggccga gagaggcggc ctgagcgaac
     7561 tggataaggc cggcttcatc aagagacagc tggtggaaac ccggcagatc acaaagcacg
     7621 tggcacagat cctggactcc cggatgaaca ctaagtacga cgagaatgac aagctgatcc
     7681 gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc cgatttccgg aaggatttcc
     7741 agttttacaa agtgcgcgag atcaacaact accaccacgc ccacgacgcc tacctgaacg
     7801 ccgtcgtggg aaccgccctg atcaaaaagt accctaagct ggaaagcccc ttcgtgtacg
     7861 gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca
     7921 aggctaccgc caagtacttc ttctacagca acatcatgaa ctttttcaag accgagatta
     7981 ccctggccaa cggcgagatc cggaagcggc ctctgatcga gacaaacggc gaaaccgggg
     8041 agatcgtgtg ggataagggc cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc
     8101 aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa gagtctatcc
     8161 tgcccaagag gaacagcgat aagctgatcg ccagaaagaa ggactgggac cctaagaagt
     8221 acggcggctt cgacagcccc accgtggcct attctgtgct ggtggtggcc aaagtggaaa
     8281 agggcaagtc caagaaactg aagagtgtga aagagctact ggggatcacc atcatggaaa
     8341 gaagcagctt cgagaagaat cccatcgact ttctggaagc caagggctac aaagaagtga
     8401 aaaaggacct gatcatcaag ctgcctaagt actccctgtt cgagctggaa aacggccgga
     8461 agagaatgct ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc ctgccctcca
     8521 aatatgtgaa cttcctgtac ctggccagcc actatgagaa gctgaagggc tcccccgagg
     8581 ataatgagca gaaacagctg tttgtggaac agcacaagca ctacctggac gagatcatcg
     8641 agcagatcag cgagttctcc aagagagtga tcctggccga cgctaatctg gacaaagtgc
     8701 tgtccgccta caacaagcac cgggataagc ccatcagaga gcaggccgag aatatcatcc
     8761 acctgtttac cctgaccaat ctgggagccc ctgccgcctt caagtacttt gacaccacca
     8821 tcgaccggaa gaggtacacc agcaccaaag aggtgctgga cgccaccctg atccaccaga
     8881 gcatcaccgg cctgtacgag acacggatcg acctgtctca gctgggaggc gacaagcgac
     8941 ctgccgccac aaagaaggct ggacaggcta agaagaagaa agattacaaa gacgatgacg
     9001 ataagggatc cggcgcaaca aacttctctc tgctgaaaca agccggagat gtcgaagaga
     9061 atcctggacc gatggccaag cctttgtctc aagaagaatc caccctcatt gaaagagcaa
     9121 cggctacaat caacagcatc cccatctctg aagactacag cgtcgccagc gcagctctct
     9181 ctagcgacgg ccgcatcttc actggtgtca atgtatatca ttttactggg ggaccttgtg
     9241 cagaactcgt ggtgctgggc actgctgctg ctgcggcagc tggcaacctg acttgtatcg
     9301 tcgcgatcgg aaatgagaac aggggcatct tgagcccctg cggacggtgc cgacaggtgc
     9361 ttctcgatct gcatcctggg atcaaagcca tagtgaagga cagtgatgga cagccgacgg
     9421 cagttgggat tcgtgaattg ctgccctctg gttatgtgtg ggagggctaa gaattcgata
     9481 tcaagcttat cggtaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     9541 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     9601 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     9661 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     9721 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     9781 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     9841 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     9901 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     9961 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
    10021 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
    10081 cgcatcgata ccgtcgacct cgagacctag aaaaacatgg agcaatcaca agtagcaata
    10141 cagcagctac caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt
    10201 ttccagtcac acctcaggta cctttaagac caatgactta caaggcagct gtagatctta
    10261 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag
    10321 atatccttga tctgtggatc taccacacac aaggctactt ccctgattgg cagaactaca
    10381 caccagggcc agggatcaga tatccactga cctttggatg gtgctacaag ctagtaccag
    10441 ttgagcaaga gaaggtagaa gaagccaatg aaggagagaa cacccgcttg ttacaccctg
    10501 tgagcctgca tgggatggat gacccggaga gagaagtatt agagtggagg tttgacagcc
    10561 gcctagcatt tcatcacatg gcccgagagc tgcatccgga ctgtactggg tctctctggt
    10621 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
    10681 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
    10741 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagg gcccgtttaa
    10801 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
    10861 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
    10921 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
    10981 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
    11041 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
    11101 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
    11161 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
    11221 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
    11281 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
    11341 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
    11401 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
    11461 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
    11521 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
    11581 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
    11641 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
    11701 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
    11761 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
    11821 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
    11881 agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
    11941 gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt
    12001 cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact tcgtggagga
    12061 cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg tccaggacca
    12121 ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc
    12181 cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga
    12241 gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt
    12301 gcacttcgtg gccgaggagc aggactgaca cgtgctacga gatttcgatt ccaccgccgc
    12361 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
    12421 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
    12481 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
    12541 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
    12601 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
    12661 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
    12721 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
    12781 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgt
//
