LOCUS       sequence-322588-       14074 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LentiCRISPRv2 variant with modified sgRNA scaffold from Chen et al. 
            2013 Cell..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14074)
  AUTHORS   Adelmann CH, Traunbauer AK, Chen B, Condon KJ, Chan SH, Kunchok T, 
            Lewis CA, Sabatini DM
  TITLE     MFSD12 mediates the import of cysteine into melanosomes and 
            lysosomes.
  JOURNAL   Nature. 2020 Nov 18. pii: 10.1038/s41586-020-2937-x. doi: 
            10.1038/s41586-020-2937-x.
  PUBMED    33208952
REFERENCE   2  (bases 1 to 14074)
  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: "Nature. 
            2020 Nov 18. pii: 10.1038/s41586-020-2937-x. doi: 
            10.1038/s41586-020-2937-x."
COMMENT     Sequence Label: LentiCRISPRv2-Opti
FEATURES             Location/Qualifiers
     source          1..14074
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1443..1676
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1861..1905
                     /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    2203..2320
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2371..2619
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2371..2391
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2542..2561
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        3839..4050
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      4069..4078
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4075..8178
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             4075..8178
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             8179..8226
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8227..8250
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             8260..8316
                     /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             8317..8913
                     /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     8810..8830
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8929..9517
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8982..9002)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9400..9411)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9589..9822
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9848..9865)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9854..10078
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10124..10552
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10211..10230)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10421..10442
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10561..10581)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10566..10895
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10746..10881
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10808..10827
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10943..10990
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11009..11383
                     /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    11513..11634
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11550..11569)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11604..11623
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11683..11699)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11683..11699)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11696..11718)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11707..11723
                     /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(11731..11737,11738..11755,11756..11761))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11776..11797
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11914..11931)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12085..12673)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12165..12184)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12844..13635,13636..13704))
                     /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     13467..13486
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13705..13809)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(13884..13903)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2341 cagagatcca gtttggttaa ttaaggtacc gagggcctat ttcccatgat tccttcatat
     2401 ttgcatatac gatacaaggc tgttagagag ataattagaa ttaatttgac tgtaaacaca
     2461 aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt
     2521 ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat
     2581 ttcttggctt tatatatctt gtggaaagga cgaaacaccg gagacggttg taaatgagca
     2641 cacaaaatac acatgctaaa atattatatt ctatgacctt tataaaatca accaaaatct
     2701 tctttttaat aactttagta tcaataatta gaatttttat gttccttttt gcaaactttt
     2761 aataaaaatg agcaaaataa aaaaacgcta gttttagtaa ctcgcgttgt tttcttcacc
     2821 tttaataata gctactccac cacttgttcc taagcggtca gctcctgctt caatcatttt
     2881 ttgagcatct tcaaatgttc taactccacc agctgcttta actaaagcat tgtctttaac
     2941 aactgacttc attagtttaa catcttcaaa tgttgcacct gattttgaaa atcctgttga
     3001 tgttttaaca aattctaatc cagcttcaac agctatttca caagctttca tgatttcttc
     3061 ttttgttaat aaacaatttt ccataataca tttaacaaca tgtgatccag ctgctttttt
     3121 tacagctttc atgtcttcta aaactaattc ataatttttg tcttttaatg caccaatatt
     3181 taataccata tcaatttctg ttgcaccatc tttaattgct tcagaaactt cgaatgcttt
     3241 tgtagctgtt gtgcatgcac ctagaggaaa acctacaaca tttgttattc ctacatttgt
     3301 gccttttaat aattctttac aatagcttgt tcaatatgaa ttaacacaaa ctgttgcaaa
     3361 atcaaattca attgcttcat cacataattg tttaatttca gctttcgtag catcttgttt
     3421 taataatgtg tgatctatat atttgtttag tttcattttt tctcctatat attcattttt
     3481 aattttaatt ctttaataat ttcgtctact ttaactttag cgttttgaac agattcacca
     3541 acacctataa aataaatttt tagtttaggt tcagttccac ttgggcgaac agcaaatcat
     3601 gacttatctt ctaaataaaa ttttagtaag tcttgtcctg gcatattata cattccatcg
     3661 atgtagttct tgctcttttt tgtacgtctc tgtttaagag ctatgctgga aacagcatag
     3721 caagtttaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt
     3781 tttgaattcg ctagctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg
     3841 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgatcc
     3901 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
     3961 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
     4021 tttcgcaacg ggtttgccgc cagaacacag gaccggttct agagcgctgc caccatggac
     4081 aagaagtaca gcatcggcct ggacatcggc accaactctg tgggctgggc cgtgatcacc
     4141 gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc
     4201 atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc
     4261 cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg
     4321 caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa
     4381 gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc
     4441 gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg
     4501 gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc
     4561 aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac
     4621 aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac
     4681 gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg
     4741 gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt
     4801 gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc
     4861 aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc
     4921 ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg
     4981 agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc
     5041 aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag
     5101 ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac
     5161 attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag
     5221 atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag
     5281 cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt
     5341 ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag
     5401 atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc
     5461 gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg
     5521 gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg
     5581 cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac
     5641 gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc
     5701 gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag
     5761 cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc
     5821 gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag
     5881 gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc
     5941 ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg
     6001 ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg
     6061 agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc
     6121 ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg
     6181 acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag
     6241 cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag
     6301 gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa
     6361 atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag
     6421 cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa
     6481 aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg
     6541 tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg
     6601 cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag
     6661 aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac
     6721 tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag
     6781 gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg
     6841 gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag
     6901 tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg
     6961 gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac
     7021 cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct
     7081 aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc
     7141 gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc
     7201 atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg
     7261 atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc
     7321 gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca
     7381 ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga
     7441 aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct
     7501 gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag
     7561 ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg
     7621 gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc
     7681 ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag
     7741 ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat
     7801 gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac
     7861 aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg
     7921 gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc
     7981 agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc
     8041 gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg
     8101 ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg
     8161 tctcagctgg gaggcgacaa gcgacctgcc gccacaaaga aggctggaca ggctaagaag
     8221 aagaaagatt acaaagacga tgacgataag ggatccggcg caacaaactt ctctctgctg
     8281 aaacaagccg gagatgtcga agagaatcct ggaccgaccg agtacaagcc cacggtgcgc
     8341 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
     8401 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
     8461 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     8521 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     8581 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     8641 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcgga
     8701 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     8761 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     8821 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     8881 acctggtgca tgacccgcaa gcccggtgcc tgaacgcgtt aagtcgacaa tcaacctctg
     8941 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     9001 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     9061 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     9121 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     9181 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     9241 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     9301 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
     9361 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     9421 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     9481 cagacgagtc ggatctccct ttgggccgcc tccccgcgtc gactttaaga ccaatgactt
     9541 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
     9601 ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc
     9661 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     9721 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     9781 gatccctcag acccttttag tcagtgtgga aaatctctag cagggcccgt ttaaacccgc
     9841 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     9901 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     9961 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
    10021 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
    10081 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
    10141 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
    10201 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
    10261 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
    10321 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
    10381 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
    10441 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
    10501 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
    10561 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
    10621 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
    10681 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
    10741 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
    10801 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
    10861 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
    10921 ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt
    10981 ataatacgac aaggtgagga actaaaccat ggccaagttg accagtgccg ttccggtgct
    11041 caccgcgcgc gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttctcccg
    11101 ggacttcgtg gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag
    11161 cgcggtccag gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct
    11221 ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg
    11281 gccggccatg accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc
    11341 ggccggcaac tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc tacgagattt
    11401 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
    11461 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
    11521 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
    11581 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
    11641 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
    11701 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
    11761 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
    11821 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
    11881 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
    11941 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
    12001 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
    12061 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
    12121 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
    12181 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
    12241 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
    12301 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
    12361 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
    12421 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
    12481 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
    12541 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
    12601 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
    12661 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
    12721 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
    12781 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
    12841 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
    12901 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
    12961 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
    13021 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
    13081 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
    13141 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
    13201 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
    13261 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
    13321 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
    13381 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
    13441 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
    13501 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
    13561 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
    13621 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
    13681 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
    13741 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
    13801 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
    13861 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
    13921 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
    13981 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
    14041 ctgcttcgcg atgtacgggc cagatatacg cgtt
//
