LOCUS       sequence-237289-       14285 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The plasmid codes for a Flag-spCas9 protein, a iRFP670 fluorescent 
            protein and a puromycin resistance. The plasmid has two BsmBI 
            acceptor sites to insert gRNA expressing sequences..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14285)
  TITLE     Lenti spCas9 T2A iRFP670 P2A puro
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 14285)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pLenti spCas9 T2A iRFP670 P2A puro
FEATURES             Location/Qualifiers
     source          1..14285
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(2594..2914)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGATTVSY
                     LN"
     primer_bind     2764..2786
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2778..2795
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2779..2795
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2805..2824
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2805..2823
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2871..2891)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2871..2889)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2910..2926)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2934..2950
                     /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(2958..2964,2965..2982,2983..2988))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3003..3024
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3122..3333
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3341..3350
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3350..3415
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3422..3442
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3467..7567
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             3467..7567
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7568..7615
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7622..7675
                     /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             7676..7726
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     regulatory      7742..7751
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7748..8680
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 670 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP670
                     /note="derived from Rhodopseudomonas palustris RpBphP6"
                     /translation="MARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAG
                     AFFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLH
                     RHDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVM
                     LYRFADDGSGMVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSR
                     IVPEHDASGAALDLSFAHLRSISPCHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYE
                     PRAVPMAQRVAAEMFADFLSLHFTAAHHQR"
     CDS             8708..8764
                     /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             8765..9361
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     9258..9278
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9377..9965
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9430..9450)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9848..9859)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10037..10270
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10296..10313)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10302..10526
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10572..11000
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10659..10678)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10869..10890
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11009..11029)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11014..11343
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11194..11329
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11256..11275
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11391..11438
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11457..11831
                     /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    11961..12082
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11998..12017)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12052..12071
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12131..12147)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    12155..12171
                     /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(12179..12185,12186..12203,12204..12209))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12224..12245
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12362..12379)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12533..13121)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12613..12632)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13292..14083,14084..14152))
                     /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     13915..13934
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14153..14257)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta agataagaga cggtggtggc gccgctacag ggcgcgtccc
     2641 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     2701 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     2761 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat
     2821 agggcgaatt gggtaccggg ccccccctcg aggtcctcca gcttttgttc cctttagtga
     2881 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     2941 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     3001 taatgagtga gctaactcac attaattgcg ttgcgctcaa tgcccgcttt ccactgcatg
     3061 acgtctccac aatctagcta ggtcttgaaa ggagtgggaa ttggctccgg tgcccgtcag
     3121 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
     3181 tccggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
     3241 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
     3301 ctttttcgca acgggtttgc cgccagaaca caggaccggt gccaccatgg actataagga
     3361 ccacgacgga gactacaagg atcatgatat tgattacaaa gacgatgacg ataagatggc
     3421 cccaaagaag aagcggaagg tcggtatcca cggagtccca gcagccgaca agaagtacag
     3481 catcggcctg gacatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa
     3541 ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa
     3601 cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag
     3661 aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt
     3721 cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag agtccttcct
     3781 ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt
     3841 ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac
     3901 cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg
     3961 ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat
     4021 ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt
     4081 ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat
     4141 cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg ccctgagcct
     4201 gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct
     4261 gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta
     4321 cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct
     4381 gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga
     4441 cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa
     4501 gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg
     4561 agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac
     4621 cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga
     4681 caacggcagc atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca
     4741 ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt
     4801 ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac
     4861 cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc
     4921 ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa
     4981 ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa
     5041 agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa
     5101 ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga
     5161 ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg
     5221 gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt
     5281 cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt
     5341 tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa
     5401 agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct
     5461 gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga
     5521 cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga
     5581 ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa
     5641 tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga
     5701 gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga
     5761 gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga
     5821 gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct
     5881 gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca
     5941 ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc ctcagagctt
     6001 tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga accggggcaa
     6061 gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct
     6121 gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg
     6181 cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca
     6241 gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa
     6301 tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt
     6361 ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga
     6421 cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag
     6481 cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga
     6541 gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt
     6601 caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa
     6661 cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt
     6721 gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag
     6781 caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa agaaggactg
     6841 ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg tgctggtggt
     6901 ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat
     6961 caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg
     7021 ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct
     7081 ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact
     7141 ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa
     7201 gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct
     7261 ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa
     7321 tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc
     7381 cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta
     7441 ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc tggacgccac
     7501 cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg
     7561 aggcgacaaa aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga aaaaggaatt
     7621 cgagggcaga ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg gcccaatggg
     7681 cgtggccgac ttgatcaaga agttcgagtc catctccaag gaggaggggg atccaccggt
     7741 cgccaccatg gcgcgtaagg tcgatctcac ctcctgcgat cgcgagccga tccacatccc
     7801 cggcagcatt cagccgtgcg gctgcctgct agcctgcgac gcgcaggcgg tgcggatcac
     7861 gcgcattacg gaaaatgccg gcgcgttctt tggacgcgaa actccgcggg tcggtgagct
     7921 actcgccgat tacttcggcg agaccgaagc ccatgcgctg cgcaacgcac tggcgcagtc
     7981 ctccgatcca aagcgaccgg cgctgatctt cggttggcgc gacggcctga ccggccgcac
     8041 cttcgacatc tcactgcatc gccatgacgg tacatcgatc atcgagttcg agcctgcggc
     8101 ggccgaacag gccgacaatc cgctgcggct gacgcggcag atcatcgcgc gcaccaaaga
     8161 actgaagtcg ctcgaagaga tggccgcacg ggtgccgcgc tatctgcagg cgatgctcgg
     8221 ctatcaccgc gtgatgttgt accgcttcgc ggacgacggc tccgggatgg tgatcggcga
     8281 ggcgaagcgc agcgacctcg agagctttct cggtcagcac tttccggcgt cgctggtccc
     8341 gcagcaggcg cggctactgt acttgaagaa cgcgatccgc gtggtctcgg attcgcgcgg
     8401 catcagcagc cggatcgtgc ccgagcacga cgcctccggc gccgcgctcg atctgtcgtt
     8461 cgcgcacctg cgcagcatct cgccctgcca tctcgaattt ctgcggaaca tgggcgtcag
     8521 cgcctcgatg tcgctgtcga tcatcattga cggcacgcta tggggattga tcatctgtca
     8581 tcattacgag ccgcgtgccg tgccgatggc gcagcgcgtc gcggccgaaa tgttcgccga
     8641 cttcttatcg ctgcacttca ccgccgccca ccaccaacgc tctagaagcg ctggtaccgg
     8701 atccggcgca acaaacttct ctctgctgaa acaagccgga gatgtcgaag agaatcctgg
     8761 accgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt
     8821 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga
     8881 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     8941 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     9001 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     9061 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     9121 gcccgcgtgg ttcctggcca ccgtcggagt ctcgcccgac caccagggca agggtctggg
     9181 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     9241 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     9301 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
     9361 aacgcgttaa gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     9421 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     9481 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     9541 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     9601 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     9661 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     9721 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     9781 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     9841 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     9901 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     9961 cccgcgtcga ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
    10021 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt
    10081 gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
    10141 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
    10201 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
    10261 atctctagca gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
    10321 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
    10381 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
    10441 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
    10501 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
    10561 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
    10621 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
    10681 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
    10741 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
    10801 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
    10861 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
    10921 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
    10981 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
    11041 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
    11101 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
    11161 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
    11221 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
    11281 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
    11341 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt
    11401 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg
    11461 ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt
    11521 tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg
    11581 tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca
    11641 ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg
    11701 tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt
    11761 gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg
    11821 agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg
    11881 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
    11941 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
    12001 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
    12061 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
    12121 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
    12181 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
    12241 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
    12301 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
    12361 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
    12421 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
    12481 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
    12541 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
    12601 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
    12661 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
    12721 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
    12781 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
    12841 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
    12901 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
    12961 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
    13021 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
    13081 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
    13141 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
    13201 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
    13261 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
    13321 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
    13381 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
    13441 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
    13501 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
    13561 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
    13621 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
    13681 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
    13741 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
    13801 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
    13861 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
    13921 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
    13981 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
    14041 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
    14101 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
    14161 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
    14221 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
    14281 ctgac
//
