LOCUS       sequence-332221-       14991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiCRISPRv2 variant with mScarlet-I fluorescence marker..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14991)
  AUTHORS   Chan W, Gottschalk RA, Yao Y, Pomerantz JL, Germain RN
  TITLE     Efficient Immune Cell Genome Engineering with Enhanced CRISPR 
            Editing Tools.
  JOURNAL   Immunohorizons. 2021 Feb 23;5(2):117-132. doi: 
            10.4049/immunohorizons.2000082.
  PUBMED    33622708
REFERENCE   2  (bases 1 to 14991)
  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"; doi: 
            "10.4049/immunohorizons"; journalName: "Immunohorizons"; date: 
            "2021-02-23- 23"; volume: "5"; issue: "2"; pages: "117-132"
COMMENT     Sequence Label: LentiCRISPRv2R
FEATURES             Location/Qualifiers
     source          1..14991
                     /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"
     misc_RNA        4501..4576
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4638..4849
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      4868..4877
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4874..8977
                     /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             4874..8977
                     /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             8978..9025
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9026..9049
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             9059..9115
                     /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             9116..9808
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="VSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGGA
                     VTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKM
                     ALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHS
                     TGGMDELYK"
     primer_bind     complement(9246..9264)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     9514..9533
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    9846..10434
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9899..9919)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10317..10328)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10506..10739
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10765..10782)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10771..10995
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11041..11469
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11128..11147)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11338..11359
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11478..11498)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11483..11812
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11663..11798
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11725..11744
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11860..11907
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11926..12300
                     /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    12430..12551
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12467..12486)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12521..12540
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12600..12616)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12600..12616)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12613..12635)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12624..12640
                     /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(12648..12654,12655..12672,12673..12678))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12693..12714
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12831..12848)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13002..13590)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13082..13101)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13761..14552,14553..14621))
                     /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     14384..14403
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14622..14726)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(14801..14820)
                     /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 atgtagtctt caacattaac aactttaagt ccagcaattt gagttaaggg tgttgctctc
     3721 aatgatttca ttaatggttc aatttttaat ttcttttctt ctggtttaaa attcaagttt
     3781 aaagtgaaag tgtaatatgc acccatttct ttaaataaat cttctaaata gtctactaat
     3841 gttttatttt gttttttata aaatcaagca gcctctgcta ttaatataga agcttgtatt
     3901 ccatctttat ctctagctga gtcatcaatt acatatccat aactttcttc ataagcaaaa
     3961 acaaaattta atccgttatc ttcttcttta gcaatttctc tacccattca tttaaatcca
     4021 gttaaagttt ttacaatatt aactccatat ttttcatgag cgattctatc acccaaatca
     4081 cttgttacaa aacttgaata tagagccgga ttttttggaa tgctatttaa gcgttttaga
     4141 tttgataatt ttcaatcaat taaaattggt cctgtttgat ttccatctaa tcttacaaaa
     4201 tgaccatcat gttttattgc cattccaaat ctgtcagcat ctgggtcatt cataataata
     4261 atatctgcat catgtttaat accatattca agcggtattt ttcatgcagg atcaaattct
     4321 ggatttggat ttacaacatt tttaaatgtt tcatcttcaa atgcatgctc ttcaacctca
     4381 ataacgttat atcctgattc acgtaatatt tttggggtaa atttagttcc tgttccatta
     4441 actgcgctaa aaataatttt taaatctttt ttagcttctt gctctttttt gtacgtctct
     4501 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
     4561 ggcaccgagt cggtgctttt ttgaattcgc tagctaggtc ttgaaaggag tgggaattgg
     4621 ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg
     4681 aggggtcggc aattgatccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga
     4741 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt
     4801 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg accggttcta
     4861 gagcgctgcc accatggaca agaagtacag catcggcctg gacatcggca ccaactctgt
     4921 gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc aagaaattca aggtgctggg
     4981 caacaccgac cggcacagca tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg
     5041 cgaaacagcc gaggccaccc ggctgaagag aaccgccaga agaagataca ccagacggaa
     5101 gaaccggatc tgctatctgc aagagatctt cagcaacgag atggccaagg tggacgacag
     5161 cttcttccac agactggaag agtccttcct ggtggaagag gataagaagc acgagcggca
     5221 ccccatcttc ggcaacatcg tggacgaggt ggcctaccac gagaagtacc ccaccatcta
     5281 ccacctgaga aagaaactgg tggacagcac cgacaaggcc gacctgcggc tgatctatct
     5341 ggccctggcc cacatgatca agttccgggg ccacttcctg atcgagggcg acctgaaccc
     5401 cgacaacagc gacgtggaca agctgttcat ccagctggtg cagacctaca accagctgtt
     5461 cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag gccatcctgt ctgccagact
     5521 gagcaagagc agacggctgg aaaatctgat cgcccagctg cccggcgaga agaagaatgg
     5581 cctgttcgga aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt
     5641 cgacctggcc gaggatgcca aactgcagct gagcaaggac acctacgacg acgacctgga
     5701 caacctgctg gcccagatcg gcgaccagta cgccgacctg tttctggccg ccaagaacct
     5761 gtccgacgcc atcctgctga gcgacatcct gagagtgaac accgagatca ccaaggcccc
     5821 cctgagcgcc tctatgatca agagatacga cgagcaccac caggacctga ccctgctgaa
     5881 agctctcgtg cggcagcagc tgcctgagaa gtacaaagag attttcttcg accagagcaa
     5941 gaacggctac gccggctaca ttgacggcgg agccagccag gaagagttct acaagttcat
     6001 caagcccatc ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc tgaacagaga
     6061 ggacctgctg cggaagcagc ggaccttcga caacggcagc atcccccacc agatccacct
     6121 gggagagctg cacgccattc tgcggcggca ggaagatttt tacccattcc tgaaggacaa
     6181 ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccctctggc
     6241 caggggaaac agcagattcg cctggatgac cagaaagagc gaggaaacca tcaccccctg
     6301 gaacttcgag gaagtggtgg acaagggcgc ttccgcccag agcttcatcg agcggatgac
     6361 caacttcgat aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga
     6421 gtacttcacc gtgtataacg agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa
     6481 gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa
     6541 ccggaaagtg accgtgaagc agctgaaaga ggactacttc aagaaaatcg agtgcttcga
     6601 ctccgtggaa atctccggcg tggaagatcg gttcaacgcc tccctgggca cataccacga
     6661 tctgctgaaa attatcaagg acaaggactt cctggacaat gaggaaaacg aggacattct
     6721 ggaagatatc gtgctgaccc tgacactgtt tgaggacaga gagatgatcg aggaacggct
     6781 gaaaacctat gcccacctgt tcgacgacaa agtgatgaag cagctgaagc ggcggagata
     6841 caccggctgg ggcaggctga gccggaagct gatcaacggc atccgggaca agcagtccgg
     6901 caagacaatc ctggatttcc tgaagtccga cggcttcgcc aacagaaact tcatgcagct
     6961 gatccacgac gacagcctga cctttaaaga ggacatccag aaagcccagg tgtccggcca
     7021 gggcgatagc ctgcacgagc acattgccaa tctggccggc agccccgcca ttaagaaggg
     7081 catcctgcag acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc
     7141 cgagaacatc gtgatcgaaa tggccagaga gaaccagacc acccagaagg gacagaagaa
     7201 cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa gagctgggca gccagatcct
     7261 gaaagaacac cccgtggaaa acacccagct gcagaacgag aagctgtacc tgtactacct
     7321 gcagaatggg cgggatatgt acgtggacca ggaactggac atcaaccggc tgtccgacta
     7381 cgatgtggac catatcgtgc ctcagagctt tctgaaggac gactccatcg acaacaaggt
     7441 gctgaccaga agcgacaaga accggggcaa gagcgacaac gtgccctccg aagaggtcgt
     7501 gaagaagatg aagaactact ggcggcagct gctgaacgcc aagctgatta cccagagaaa
     7561 gttcgacaat ctgaccaagg ccgagagagg cggcctgagc gaactggata aggccggctt
     7621 catcaagaga cagctggtgg aaacccggca gatcacaaag cacgtggcac agatcctgga
     7681 ctcccggatg aacactaagt acgacgagaa tgacaagctg atccgggaag tgaaagtgat
     7741 caccctgaag tccaagctgg tgtccgattt ccggaaggat ttccagtttt acaaagtgcg
     7801 cgagatcaac aactaccacc acgcccacga cgcctacctg aacgccgtcg tgggaaccgc
     7861 cctgatcaaa aagtacccta agctggaaag cgagttcgtg tacggcgact acaaggtgta
     7921 cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta
     7981 cttcttctac agcaacatca tgaacttttt caagaccgag attaccctgg ccaacggcga
     8041 gatccggaag cggcctctga tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa
     8101 gggccgggat tttgccaccg tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa
     8161 aaagaccgag gtgcagacag gcggcttcag caaagagtct atcctgccca agaggaacag
     8221 cgataagctg atcgccagaa agaaggactg ggaccctaag aagtacggcg gcttcgacag
     8281 ccccaccgtg gcctattctg tgctggtggt ggccaaagtg gaaaagggca agtccaagaa
     8341 actgaagagt gtgaaagagc tgctggggat caccatcatg gaaagaagca gcttcgagaa
     8401 gaatcccatc gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat
     8461 caagctgcct aagtactccc tgttcgagct ggaaaacggc cggaagagaa tgctggcctc
     8521 tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc tccaaatatg tgaacttcct
     8581 gtacctggcc agccactatg agaagctgaa gggctccccc gaggataatg agcagaaaca
     8641 gctgtttgtg gaacagcaca agcactacct ggacgagatc atcgagcaga tcagcgagtt
     8701 ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa
     8761 gcaccgggat aagcccatca gagagcaggc cgagaatatc atccacctgt ttaccctgac
     8821 caatctggga gcccctgccg ccttcaagta ctttgacacc accatcgacc ggaagaggta
     8881 caccagcacc aaagaggtgc tggacgccac cctgatccac cagagcatca ccggcctgta
     8941 cgagacacgg atcgacctgt ctcagctggg aggcgacaag cgacctgccg ccacaaagaa
     9001 ggctggacag gctaagaaga agaaagatta caaagacgat gacgataagg gatccggcgc
     9061 aacaaacttc tctctgctga aacaagccgg agatgtcgaa gagaatcctg gaccggtgag
     9121 caagggcgag gcagtgatca aggagttcat gcggttcaag gtgcacatgg agggctccat
     9181 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
     9241 gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttctcctggg acatcctgtc
     9301 ccctcagttc atgtacggct ccagggcctt catcaagcac cccgccgaca tccccgacta
     9361 ctataagcag tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
     9421 cggcgccgtg accgtgaccc aggacacctc cctggaggac ggcaccctga tctacaaggt
     9481 gaagctccgc ggcaccaact tccctcctga cggccccgta atgcagaaga agacaatggg
     9541 ctgggaagcg tccaccgagc ggttgtaccc cgaggacggc gtgctgaagg gcgacattaa
     9601 gatggccctg cgcctgaagg acggcggccg ctacctggcg gacttcaaga ccacctacaa
     9661 ggccaagaag cccgtgcaga tgcccggcgc ctacaacgtc gaccgcaagt tggacatcac
     9721 ctcccacaac gaggactaca ccgtggtgga acagtacgaa cgctccgagg gccgccactc
     9781 caccggcggc atggacgagc tgtacaagta actgataata gtgataatag tcgcgttaag
     9841 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     9901 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     9961 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
    10021 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
    10081 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
    10141 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
    10201 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
    10261 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
    10321 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
    10381 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcgtcgac
    10441 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg
    10501 gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg
    10561 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
    10621 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
    10681 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
    10741 ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
    10801 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
    10861 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
    10921 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
    10981 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
    11041 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
    11101 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
    11161 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
    11221 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
    11281 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
    11341 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
    11401 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
    11461 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
    11521 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
    11581 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
    11641 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
    11701 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
    11761 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
    11821 gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca
    11881 tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc
    11941 agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac
    12001 cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac
    12061 gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg
    12121 gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac
    12181 ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag
    12241 ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga
    12301 cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
    12361 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
    12421 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
    12481 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
    12541 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
    12601 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
    12661 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
    12721 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
    12781 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
    12841 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
    12901 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
    12961 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
    13021 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
    13081 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
    13141 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
    13201 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
    13261 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
    13321 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
    13381 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
    13441 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
    13501 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
    13561 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
    13621 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
    13681 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
    13741 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
    13801 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
    13861 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
    13921 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
    13981 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
    14041 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
    14101 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
    14161 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
    14221 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
    14281 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
    14341 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
    14401 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
    14461 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
    14521 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
    14581 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
    14641 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
    14701 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
    14761 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
    14821 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
    14881 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
    14941 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt t
//
