LOCUS       sequence-372267-       15182 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentivirus transfer plasmid encoding Cas9, puromycin resistance, and
            4 sgRNAs targeting human safe targeting loci.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15182)
  AUTHORS   Brewer WJ, Xet-Mull AM, Yu A, Sweeney MI, Walton EM, Tobin DM
  TITLE     Macrophage NFATC2 mediates angiogenic signaling during mycobacterial
            infection.
  JOURNAL   Cell Rep. 2022 Dec 13;41(11):111817. doi: 
            10.1016/j.celrep.2022.111817.
  PUBMED    36516756
REFERENCE   2  (bases 1 to 15182)
  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: "Cell Rep.";
            date: "2022-12-13"; volume: "41(11):111817. doi"; pages: " 
            10.1016/j.celrep.2022.111817"
COMMENT     Sequence Label: pLV hUbC-Cas9-P2A-Puro_ST-sgRNA
FEATURES             Location/Qualifiers
     source          1..15182
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..199
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     154..174
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             217..397
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    444..569
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1062..1295
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1480..1524
                     /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    1822..1939
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(1984..2017)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2022..2335
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     2254..2274
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     misc_RNA        2356..2431
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        complement(2461..2536)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(2556..2804)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(2614..2633)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(2784..2804)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     promoter        2834..3048
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     2969..2988
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     misc_RNA        3076..3151
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        complement(3181..3256)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3551..4762
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4599..4618
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      4796..4805
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4802..8905
                     /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             4802..8905
                     /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             8906..8953
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8954..8977
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             8987..9043
                     /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             9044..9640
                     /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     9537..9557
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9656..10244
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9709..9729)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10127..10138)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10316..10549
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10575..10592)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10581..10805
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10851..11279
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10938..10957)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11148..11169
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11288..11308)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11293..11622
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11473..11608
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11535..11554
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11670..11717
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11736..12110
                     /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    12240..12361
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12277..12296)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12331..12350
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12410..12426)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12410..12426)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12423..12445)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12434..12450
                     /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(12458..12464,12465..12482,12483..12488))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12503..12524
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12641..12658)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12812..13400)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12892..12911)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13571..14362,14363..14431))
                     /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     14194..14213
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14432..14536)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(14611..14630)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        14802..15181
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
       61 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      121 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      181 gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc
      241 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
      301 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
      361 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
      421 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
      481 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
      541 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
      601 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
      661 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
      721 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
      781 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
      841 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
      901 agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
      961 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     1021 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     1081 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     1141 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     1201 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     1261 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     1321 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     1381 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     1441 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     1501 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     1561 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     1621 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     1681 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     1741 agagacagat ccattcgatt agtgaacgga tcggcactgc gtgcgccaat tctgcagaca
     1801 aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg
     1861 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat
     1921 tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag tttggttaat
     1981 taaataactt cgtatagcat acattatacg aagttatgat agatccgacg ccgccatctc
     2041 taggcccgcg ccggccccct cgcacagact tgtgggagaa gctcggctac tcccctgccc
     2101 cggttaattt gcatataata tttcctagta actatagagg cttaatgtgc gataaaagac
     2161 agataatctg ttctttttaa tactagctac attttacatg ataggcttgg atttctataa
     2221 gagatacaaa tactaaatta ttattttaaa aaacagcaca aaaggaaact caccctaact
     2281 gtaaagtaat tgtgtgtttt gagactataa atatcccttg gagaaaagcc ttgtttggct
     2341 aagtactcta acagggtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
     2401 tcaacttgaa aaagtggcac cgagtcggtg ctttttttgt acgctcagcg gagaaaaaaa
     2461 gcaccgactc ggtgccactt tttcaagttg ataacggact agccttattt taacttgcta
     2521 tttctagctc taaaactcga caactgtcac catccggtgt ttcgtccttt ccacaagata
     2581 tataaagcca agaaatcgaa atactttcaa gttacggtaa gcatatgata gtccatttta
     2641 aaacataatt ttaaaactgc aaactaccca agaaattatt actttctacg tcacgtattt
     2701 tgtactaata tctttgtgtt tacagtcaaa ttaattccaa ttatctctct aacagccttg
     2761 tatcgtatat gcaaatatga aggaatcatg ggaaataggc cctcttcctg cccgaccttg
     2821 gatcgatccg cctgaacgct gacgtcatca acccgctcca aggaatcgcg ggcccagtgt
     2881 cactaggcgg gaacacccag cgcgcgtgcg ccctggcagg aagatggctg tgagggacag
     2941 gggagtggcg ccctgcaata tttgcatgtc gctatgtgtt ctgggaaatc accataaacg
     3001 tgaaatgtct ttggatttgg gaatcttata agttctgtat gagaccactc tttcccagtg
     3061 cagttctccg ggttggtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
     3121 tcaacttgaa aaagtggcac cgagtcggtg ctttttttgg tcatcagtcc gagaaaaaaa
     3181 gcaccgactc ggtgccactt tttcaagttg ataacggact agccttattt taacttgcta
     3241 tttctagctc taaaacactc aggaagttat ccaccgaggt acccaagcgg cgcacaagct
     3301 atataaacct gaaggaagtc tcaactttac acttaggtca agttgcttat cgtactagag
     3361 cttcagcagg aaatttaact aaaatctaat ttaaccagca tagcaaatat catttattcc
     3421 caaaatgcta aagtttgaga taaacggact tgatttccgg ctgttttgac actatccaga
     3481 atgccttgca gatgggtggg gcatgctaaa tactgcagac aattaattaa cccgtgtcgg
     3541 ctccagatct ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc cctcctcacg
     3601 gcgagcgctg ccacgtcaga cgaagggcgc agcgagcgtc ctgatccttc cgcccggacg
     3661 ctcaggacag cggcccgctg ctcataagac tcggccttag aaccccagta tcagcagaag
     3721 gacattttag gacgggactt gggtgactct agggcactgg ttttctttcc agagagcgga
     3781 acaggcgagg aaaagtagtc ccttctcggc gattctgcgg agggatctcc gtggggcggt
     3841 gaacgccgat gattatataa ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg
     3901 ggatttgggt cgcggttctt gtttgtggat cgctgtgatc gtcacttggt gagtagcggg
     3961 ctgctgggct ggccggggct ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg
     4021 gagagaccgc caagggctgt agtctgggtc cgcgagcaag gttgccctga actgggggtt
     4081 ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg aatggaagac gcttgtgagg
     4141 cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga acccaaggtc
     4201 ttgaggcctt cgctaatgcg ggaaagctct tattcgggtg agatgggctg gggcaccatc
     4261 tggggaccct gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg
     4321 gggcggcagt tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc
     4381 tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccacctgcc
     4441 ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc
     4501 tctcctgaat cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt
     4561 tctttggtcg gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat
     4621 gcgctcgggg ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc
     4681 atttgggtca atatgtaatt ttcagtgtta gactagtaaa ttgtccgcta aattctggcc
     4741 gtttttggct tttttgttag acgaagcttg ggctgcaggt cgactctaga gcgctgccac
     4801 catggacaag aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt
     4861 gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg
     4921 gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga
     4981 ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg
     5041 ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag
     5101 actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg
     5161 caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa
     5221 gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca
     5281 catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga
     5341 cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc
     5401 catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag
     5461 acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa
     5521 cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga
     5581 ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc
     5641 ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat
     5701 cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc
     5761 tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg
     5821 gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc
     5881 cggctacatt gacggcggag ccagccagga agagttctac aagttcatca agcccatcct
     5941 ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg
     6001 gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca
     6061 cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat
     6121 cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag
     6181 cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga
     6241 agtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa
     6301 gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt
     6361 gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct
     6421 gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac
     6481 cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat
     6541 ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat
     6601 tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt
     6661 gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc
     6721 ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg
     6781 caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct
     6841 ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga
     6901 cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct
     6961 gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac
     7021 agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt
     7081 gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag
     7141 aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc
     7201 cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg
     7261 ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg atgtggacca
     7321 tatcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag
     7381 cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa
     7441 gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct
     7501 gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca
     7561 gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa
     7621 cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc
     7681 caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa
     7741 ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa
     7801 gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa
     7861 gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag
     7921 caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg
     7981 gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt
     8041 tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt
     8101 gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat
     8161 cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc
     8221 ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt
     8281 gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga
     8341 ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa
     8401 gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact
     8461 gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag
     8521 ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga
     8581 acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt
     8641 gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa
     8701 gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc
     8761 ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa
     8821 agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat
     8881 cgacctgtct cagctgggag gcgacaagcg acctgccgcc acaaagaagg ctggacaggc
     8941 taagaagaag aaagattaca aagacgatga cgataaggga tccggcgcaa caaacttctc
     9001 tctgctgaaa caagccggag atgtcgaaga gaatcctgga ccgaccgagt acaagcccac
     9061 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
     9121 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
     9181 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     9241 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     9301 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     9361 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     9421 cgtcggagtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     9481 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     9541 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     9601 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga acgcgttaag tcgacaatca
     9661 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     9721 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     9781 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     9841 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     9901 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     9961 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
    10021 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
    10081 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
    10141 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
    10201 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcgtcgac tttaagacca
    10261 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
    10321 gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg
    10381 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
    10441 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
    10501 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag ggcccgttta
    10561 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
    10621 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
    10681 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
    10741 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
    10801 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
    10861 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
    10921 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
    10981 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
    11041 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
    11101 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
    11161 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
    11221 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
    11281 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
    11341 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
    11401 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
    11461 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
    11521 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
    11581 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
    11641 atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg
    11701 gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc agtgccgttc
    11761 cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt
    11821 tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt
    11881 tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc
    11941 gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg
    12001 cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc
    12061 gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctac
    12121 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
    12181 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
    12241 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
    12301 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
    12361 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
    12421 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
    12481 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
    12541 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
    12601 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
    12661 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
    12721 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
    12781 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
    12841 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
    12901 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
    12961 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
    13021 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
    13081 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
    13141 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
    13201 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
    13261 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
    13321 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
    13381 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
    13441 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
    13501 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
    13561 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
    13621 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
    13681 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
    13741 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
    13801 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
    13861 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
    13921 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
    13981 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
    14041 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
    14101 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
    14161 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
    14221 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
    14281 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
    14341 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
    14401 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
    14461 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
    14521 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
    14581 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
    14641 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
    14701 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
    14761 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag
    14821 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
    14881 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
    14941 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
    15001 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
    15061 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
    15121 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
    15181 gg
//
