LOCUS       sequence-370636-       14148 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentivirus transfer plasmid encoding Cas9, puromycin resistance, and
            a BsmBI array for cloning 4 sgRNAs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14148)
  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 14148)
  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_BsmBI-sgRNA-BsmBI
FEATURES             Location/Qualifiers
     source          1..14148
                     /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)."
     CDS             complement(2004..2330)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGATTVSY
                     HNFV"
     primer_bind     2180..2202
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2194..2211
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2195..2211
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2221..2240
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2221..2239
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2287..2307)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2287..2305)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2326..2342)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2350..2366
                     /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(2374..2380,2381..2398,2399..2404))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2419..2440
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2517..3728
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3565..3584
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      3762..3771
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3768..7871
                     /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             3768..7871
                     /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             7872..7919
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7920..7943
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             7953..8009
                     /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             8010..8606
                     /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     8503..8523
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8622..9210
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8675..8695)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9093..9104)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9282..9515
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9541..9558)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9547..9771
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9817..10245
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9904..9923)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10114..10135
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10254..10274)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10259..10588
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10439..10574
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10501..10520
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10636..10683
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10702..11076
                     /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    11206..11327
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11243..11262)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11297..11316
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11376..11392)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    11400..11416
                     /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(11424..11430,11431..11448,11449..11454))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11469..11490
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11607..11624)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11778..12366)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11858..11877)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12537..13328,13329..13397))
                     /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     13160..13179
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13398..13502)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(13577..13596)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        13768..14147
                     /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 aagagacggt ggtggcgccg
     2041 ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     2101 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     2161 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg
     2221 taatacgact cactataggg cgaattgggt accgggcccc ccctcgaggt cctccagctt
     2281 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
     2341 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     2401 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     2461 cgctttccac tgcatgacgt ctccacaatt aattaacccg tgtcggctcc agatctggcc
     2521 tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc ctcacggcga gcgctgccac
     2581 gtcagacgaa gggcgcagcg agcgtcctga tccttccgcc cggacgctca ggacagcggc
     2641 ccgctgctca taagactcgg ccttagaacc ccagtatcag cagaaggaca ttttaggacg
     2701 ggacttgggt gactctaggg cactggtttt ctttccagag agcggaacag gcgaggaaaa
     2761 gtagtccctt ctcggcgatt ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt
     2821 atataaggac gcgccgggtg tggcacagct agttccgtcg cagccgggat ttgggtcgcg
     2881 gttcttgttt gtggatcgct gtgatcgtca cttggtgagt agcgggctgc tgggctggcc
     2941 ggggctttcg tggccgccgg gccgctcggt gggacggaag cgtgtggaga gaccgccaag
     3001 ggctgtagtc tgggtccgcg agcaaggttg ccctgaactg ggggttgggg ggagcgcagc
     3061 aaaatggcgg ctgttcccga gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc
     3121 gttgaaacaa ggtggggggc atggtgggcg gcaagaaccc aaggtcttga ggccttcgct
     3181 aatgcgggaa agctcttatt cgggtgagat gggctggggc accatctggg gaccctgacg
     3241 tgaagtttgt cactgactgg agaactcggt ttgtcgtctg ttgcgggggc ggcagttatg
     3301 gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga
     3361 cgtcacccgt tctgttggct tataatgcag ggtggggcca cctgccggta ggtgtgcggt
     3421 aggcttttct ccgtcgcagg acgcagggtt cgggcctagg gtaggctctc ctgaatcgac
     3481 aggcgccgga cctctggtga ggggagggat aagtgaggcg tcagtttctt tggtcggttt
     3541 tatgtaccta tcttcttaag tagctgaagc tccggttttg aactatgcgc tcggggttgg
     3601 cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt gggtcaatat
     3661 gtaattttca gtgttagact agtaaattgt ccgctaaatt ctggccgttt ttggcttttt
     3721 tgttagacga agcttgggct gcaggtcgac tctagagcgc tgccaccatg gacaagaagt
     3781 acagcatcgg cctggacatc ggcaccaact ctgtgggctg ggccgtgatc accgacgagt
     3841 acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga
     3901 agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc acccggctga
     3961 agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat ctgcaagaga
     4021 tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg gaagagtcct
     4081 tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac atcgtggacg
     4141 aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa ctggtggaca
     4201 gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg atcaagttcc
     4261 ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg gacaagctgt
     4321 tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc aacgccagcg
     4381 gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc
     4441 tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg attgccctga
     4501 gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat gccaaactgc
     4561 agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag atcggcgacc
     4621 agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg ctgagcgaca
     4681 tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg atcaagagat
     4741 acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg
     4801 agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc tacattgacg
     4861 gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa aagatggacg
     4921 gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag cagcggacct
     4981 tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc attctgcggc
     5041 ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag aagatcctga
     5101 ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga ttcgcctgga
     5161 tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg gtggacaagg
     5221 gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac ctgcccaacg
     5281 agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat aacgagctga
     5341 ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc ggcgagcaga
     5401 aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga
     5461 aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag
     5521 atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc aaggacaagg
     5581 acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg accctgacac
     5641 tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg
     5701 acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg ctgagccgga
     5761 agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat ttcctgaagt
     5821 ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc ctgaccttta
     5881 aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac gagcacattg
     5941 ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg aaggtggtgg
     6001 acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc gaaatggcca
     6061 gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg aagcggatcg
     6121 aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc
     6181 agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat atgtacgtgg
     6241 accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc gtgcctcaga
     6301 gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac aagaaccggg
     6361 gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac tactggcggc
     6421 agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc aaggccgaga
     6481 gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg gtggaaaccc
     6541 ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact aagtacgacg
     6601 agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg
     6661 atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac caccacgccc
     6721 acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg
     6781 aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg atcgccaaga
     6841 gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac atcatgaact
     6901 ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct ctgatcgaga
     6961 caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc accgtgcgga
     7021 aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct
     7081 tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc agaaagaagg
     7141 actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat tctgtgctgg
     7201 tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg
     7261 ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt ctggaagcca
     7321 agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac tccctgttcg
     7381 agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag aagggaaacg
     7441 aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac tatgagaagc
     7501 tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag cacaagcact
     7561 acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc ctggccgacg
     7621 ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc atcagagagc
     7681 aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct gccgccttca
     7741 agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag gtgctggacg
     7801 ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac ctgtctcagc
     7861 tgggaggcga caagcgacct gccgccacaa agaaggctgg acaggctaag aagaagaaag
     7921 attacaaaga cgatgacgat aagggatccg gcgcaacaaa cttctctctg ctgaaacaag
     7981 ccggagatgt cgaagagaat cctggaccga ccgagtacaa gcccacggtg cgcctcgcca
     8041 cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
     8101 ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
     8161 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
     8221 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
     8281 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc
     8341 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggagtctcgc
     8401 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg
     8461 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
     8521 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
     8581 gcatgacccg caagcccggt gcctgaacgc gttaagtcga caatcaacct ctggattaca
     8641 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     8701 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     8761 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     8821 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     8881 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     8941 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     9001 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga
     9061 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     9121 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     9181 gtcggatctc cctttgggcc gcctccccgc gtcgacttta agaccaatga cttacaaggc
     9241 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     9301 ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg
     9361 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     9421 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     9481 cagacccttt tagtcagtgt ggaaaatctc tagcagggcc cgtttaaacc cgctgatcag
     9541 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     9601 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     9661 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     9721 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     9781 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     9841 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     9901 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     9961 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
    10021 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
    10081 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
    10141 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
    10201 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
    10261 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
    10321 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
    10381 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
    10441 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
    10501 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
    10561 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
    10621 atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac
    10681 gacaaggtga ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg
    10741 cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc
    10801 gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc
    10861 caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag
    10921 ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc
    10981 atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc
    11041 aactgcgtgc acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc
    11101 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
    11161 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
    11221 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
    11281 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
    11341 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
    11401 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
    11461 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
    11521 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
    11581 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
    11641 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
    11701 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
    11761 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
    11821 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
    11881 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
    11941 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
    12001 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
    12061 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
    12121 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
    12181 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
    12241 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
    12301 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
    12361 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
    12421 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
    12481 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
    12541 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
    12601 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
    12661 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
    12721 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
    12781 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
    12841 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
    12901 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
    12961 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
    13021 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
    13081 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
    13141 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
    13201 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
    13261 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
    13321 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
    13381 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
    13441 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
    13501 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
    13561 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
    13621 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
    13681 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc
    13741 gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat taatagtaat
    13801 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
    13861 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
    13921 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
    13981 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
    14041 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
    14101 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatgg
//
